Methods for Use of a Specific Anti-Angiogenic Adenoviral Agent

Abstract

Anti-angiogenic adenovirus vectors, and therapeutic use thereof are provided, and more particularly, but not exclusively, clinical protocols for treatment of solid tumors in patients with an Ad5-PPE-1-3X-fas-chimera adenovirus vector.

Description

FIELD AND BACKGROUND OF THE INVENTION

[0001] The present invention, in some embodiments thereof, relates to anti-angiogenic adenovirus vectors, and therapeutic use thereof, and more particularly, but not exclusively, to clinical protocols for treatment of solid tumors in patients with an Ad5-PPE-1-3X-fas-chimera adenovirus vector.

[0002] Angiogenesis is a process of new blood vessel formation by sprouting from pre-existing neighboring vessels. This process is common and major feature of several pathologies. Among these are diseases in which excessive angiogenesis is a part of the pathology and thus is a target of therapy, most significantly, cancer. Angiogenesis occurs in tumors and permits their growth, invasion and metastatic proliferation. In 1971, Folkman proposed that tumor growth and metastases are angiogenesis dependent, and suggested that inhibiting angiogenesis may be a strategy to arrest tumor growth.

[0003] There are several molecules involved in angiogenesis, from cell surface molecules to transcription factors to growth factors. Hypoxia is an important environmental factor that leads to neovascularization, inducing release of several pro-angiogenic cytokines, including vascular endothelial growth factors (VEGF) and their receptors, members of the angiopoietin family, basic fibroblast growth factor, and endothelin-1 (ET-1). These factors mediate induction of angiogenesis through control of activation, proliferation and migration of endothelial cells.

[0004] Recombinant forms of endogenous inhibitors of angiogenesis have been tested for the treatment of cancer, however the potential pharmacokinetic, biotechnological and economic drawbacks of chronic delivery of these recombinant inhibitors have led scientists to develop other approaches. The development of the anti-VEGF monoclonal antibody bevacizumab has validated anti-angiogenic targeting as a complementary therapeutic modality to chemotherapy. Several small molecule inhibitors, including second-generation multi-targeted tyrosine kinase inhibitors, have also shown promise as antiangiogenic agents for cancer.

[0005] The drawbacks of chronic delivery of recombinant inhibitors, antibodies, and small molecules, as well as the limited activity manifested when these drugs are administered as monotherapy have led to the development of anti-angiogenic gene therapies. Gene therapy is an emerging modality for treating inherited and acquired human diseases. However, a number of obstacles have impeded development of successful gene therapy, including duration of expression, induction of the immune response, cytotoxicity of the vectors and tissue specificity.

[0006] Two general strategies for anti-cancer gene therapy have proposed: tumor directed or systemic gene therapy. The lack of success in targeting gene therapy products to cancerous cells or their environment by systemic treatments, and the danger of significant anti-drug or anti-vector immunity has caused most therapies to be administered to the tumor itself, despite the advantages of systemic administration. "Adenoviral vaccines", designed to induce immunity to a recombinant antigen or epitope expressed in the patient's body, have been tried but produce mostly disappointing results. Thus, elaborate, potentially dangerous and costly strategies for eluding pathological host immune responses to systemic and repeated administration of therapeutic recombinant adenoviral vectors have been proposed, including immunosuppression, oral tolerization to vector antigens and genetic modification of the vectors (see Bangari et al, Current Gene Therapy 2006; 6: 215-226).

[0007] U.S. Pat. No. 5,747,340 teaches use of a murine endothelial cell-specific promoter which shows selectivity towards angiogenic cells, and therapeutic applications thereof.

[0008] International Application WO/2008/132729 discloses a non-replicating adenovirus vector (Ad5, E1 deleted), containing a modified murine pre-proendothelin promoter (PPE-1-3X) and a fas-chimera transgene [Fas and human tumor necrosis factor (TNF) receptor] which has been developed, in which the modified murine promoter (PPE-1-3X), is able to restrict expression of the fas chimera transgene to angiogenic blood vessels, leading to targeted apoptosis of these vessels.

[0009] Endothelial-specific gene therapy with the PPE-1-3X promoter does not increase the specificity of viral interactions with the host (e.g. transfection) but restricts the expression of the transgene to those tissues that endogenously recognize the modified promoter-angiogenic endothelial cells. The chimeric receptor can trigger the Fas pathway by binding TNFα, which is less toxic in non-tumoral tissues than using the Fas/Fas ligand mechanism, which is highly expressed in non-tumoral normal tissues such as the liver. Further, TNFα was found to be abundant in the microenvironment of tumors adding to the specificity of the transgene activity in the tumor and its surroundings.

[0010] Preliminary studies have shown that a single systemic injection of a PPE-1-3X-fas chimera results in transgene expression restricted to the tumor-bearing organ, causing tumor growth retardation, necrosis of the blood vessels in the metastatic tumor mass and reduction in tumor burden in B16 melanoma and Lewis lung carcinoma mice models.

[0011] However, an effective procedure for administration of a therapeutic amount of a recombinant anti-angiogenic adenovirus vector in the clinical setting is still lacking. As such, there is a great need for defining the parameters of clinically viable protocols for anti-angiogenic-adenoviral treatment of conditions associated with neovascularization, such as cancer, without the disadvantages of the current methods as described herein.

SUMMARY OF THE INVENTION

[0012] According to an aspect of some embodiments of the present invention there is provided a method of treating a solid tumor in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a non-replicating adenovirus vector, the vector comprises a polynucleotide which comprises a fas-chimera transgene transcriptionally linked to a murine pre-proendothelin promoter, wherein the therapeutically effective amount is at least 1×10⁸ virus particles.

[0013] According to an aspect of some embodiments of the present invention there is provided a method of treating a solid tumor in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a non-replicating adenovirus vector which comprises a polynucleotide comprising a fas-chimera transgene transcriptionally linked to a murine pre-proendothelin promoter, the adenovirus vector comprising a nucleic acid sequence as set forth in SEQ ID NOs: 9 or 10.

[0014] According to an aspect of some embodiments of the present invention there is provided a method of treating a solid tumor in a subject in need thereof, the method comprising administering to the subject, in at least two separate doses, a therapeutically effective amount of a non-replicating adenovirus vector which comprises a polynucleotide comprising a fas-chimera transgene transcriptionally linked to a murine pre-proendothelin promoter, wherein the time between administration of the first dose and the at least a second dose is sufficient for anti-Ad5 antibody formation in the subject.

[0015] According to an aspect of some embodiments of the present invention there is provided a method of treating a solid tumor in a subject in need thereof, the method comprising administering to the subject a single intravenous dose of 3×10¹² or 1×10¹³ virus particles of a non-replicating adenovirus vector comprising a polynucleotide having a nucleotide sequence as set forth in SEQ ID NOs: 9 or 10.

[0016] According to an aspect of some embodiments of the present invention there is provided a method of treating a thyroid cancer in a subject in need thereof, the method comprising administering to the subject a single intravenous dose of 3×10¹² or 1×10¹³ virus particles of a non-replicating adenovirus vector comprising a polynucleotide having a nucleotide sequence as set forth in SEQ ID NOs: 9 or 10.

[0017] According to an aspect of some embodiments of the present invention there is provided a method of treating a neuroendocrine cancer in a subject in need thereof, the method comprising administering to the subject a single intravenous dose of 3×10¹² or 1×10¹² virus particles of a non-replicating adenovirus vector comprising a polynucleotide having a nucleotide sequence as set forth in SEQ ID NO: 9 or 10.

[0018] According to some embodiments of the present invention administering the dose of adenovirus vector inhibits angiogenesis of the tumor.

[0019] According to some embodiments of the present invention administering the dose of adenovirus inhibits growth of the tumor.

[0020] According to an aspect of some embodiments of the present invention there is provided a method for administering a therapeutically effective amount of a therapeutic composition comprising an adenoviral vector to a subject in need thereof comprising administering the composition to the subject at least twice, wherein the administration does not induce a dose-dependent increase in antibodies against the adenoviral vector in the subject.

[0021] According to an aspect of some embodiments of the present invention there is provided a kit for treating a solid tumor in a subject in need thereof, comprising a unit dosage of virus particles of a non-replicating adenovirus vector comprising a polynucleotide having a nucleotide sequence as set forth in SEQ ID NOs: 9 or 10, wherein the non-replicating adenovirus vector is formulated for intravenous administration, and instructions for administration of the adenovirus.

[0022] According to some embodiments of the present invention the unit dosage comprises about 3×10¹² virus particles, at least about 1×10⁸ to about 1×10¹⁶ virus particles, at least about 1×10¹¹ to about 1×10¹³ virus particles, optionally at least about 3×10¹² virus particles.

[0023] According to some embodiments of the present invention the fas-chimera transgene comprises a polynucleotide having a nucleotide sequence as set forth in SEQ ID NO: 3.

[0024] According to yet other embodiments of the present invention the fas-chimera transgene comprises a polynucleotide having a nucleotide sequence as set forth in SEQ ID NO: 2.

[0025] According to still other embodiments of the present invention the fas-chimera transgene comprises a polynucleotide having a nucleotide sequence as set forth in SEQ ID NO: 4.

[0026] According to some embodiments of the present invention the murine pre-pro endothelin promoter comprises a polynucleotide having a nucleotide sequence as set forth in SEQ ID NO: 6.

[0027] According to some embodiments of the present invention the murine pre-pro endothelin promoter comprises a polynucleotide having a nucleotide sequence as set forth in SEQ ID NO: 7.

[0028] According to yet other embodiments of the present invention the murine pre-pro endothelin promoter comprises a polynucleotide having a nucleotide sequence as set forth in SEQ ID NO: 8.

[0029] According to yet other embodiments of the present invention the murine pre-pro endothelin promoter comprises a polynucleotide having a nucleotide sequence as set forth in SEQ ID NO: 5.

[0030] According to still other embodiments of the present invention the murine prepro-endothelin promoter comprises a polynucleotide having a nucleotide sequence as set forth in SEQ ID NO: 12.

[0031] According to some embodiments of the present invention the murine pre-pro endothelin promoter comprises a polynucleotide having a nucleotide sequence as set forth in SEQ ID NO: 13.

[0032] According to some embodiments of the present invention the non-replicating adenovirus vector is an adenovirus 5 vector.

[0033] According to yet other embodiments of the present invention the adenovirus 5 vector comprises a nucleic acid sequence as set forth in SEQ ID NOs: 9 or 10.

[0034] According to some embodiments of the present invention the solid tumor is a cancerous tumor.

[0035] According to still other embodiments of the present invention the solid tumor is a primary tumor.

[0036] According to yet other embodiments of the present invention the solid tumor is a metastatic tumor.

[0037] According to some embodiments the solid tumor is a thyroid tumor.

[0038] According to some embodiments the solid tumor is a neuroendocrine tumor.

[0039] According to some embodiments administering the dose of adenovirus vector inhibits angiogenesis of the tumor.

[0040] According to some embodiments administering the dose of the adenovirus vector inhibits growth of the tumor.

[0041] According to some embodiments the adenovirus is detected in the blood of the subject at least about 4 days post administration.

[0042] According to some embodiments an amount of serum anti-adenovirus antibodies is increased following the administering, and the adenovirus is detected in the blood of the subject at least about 21 days post administration.

[0043] According to some embodiments of the present invention the adenovirus vector is administered systemically.

[0044] According to still other embodiments of the present invention the vector is administered systemically by a route selected from the group consisting of intra-articular administration, intravenous administration, intraperitoneal administration, subcutaneous administration, infusion, oral administration, rectal administration, nasal administration and inhalation.

[0045] According to still other embodiments of the present invention administering the adenovirus vector is in at least two separate systemic doses.

[0046] According to some embodiments of the present invention the administering of the adenovirus vector is in a first dose and at least a second additional dose, wherein the first dose is sufficient to induce anti-Ad5 antibodies in the subject, and wherein the time between administration of the first dose and the at least a second dose is sufficient for anti-Ad5 antibody formation in the subject

[0047] According to some embodiments of the present invention the subject is further receiving a chemotherapeutic agent as well as treatment with the virus particles of said non-replicating adenovirus vector.

[0048] According to some embodiments of the present invention the chemotherapeutic agent is administered prior to, concomitantly with, or following treatment with said virus particles.

[0049] According to some embodiments of the present invention the chemotherapeutic agent is sunitinib.

[0050] Unless otherwise defined, all technical and/or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the invention, exemplary methods and/or materials are described below. In case of conflict, the patent specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and are not intended to be necessarily limiting.

BRIEF DESCRIPTION OF THE DRAWINGS

[0051] Some embodiments of the invention are herein described, by way of example only, with reference to the accompanying drawings and images. With specific reference now to the drawings and images in detail, it is stressed that the particulars shown are by way of example and for purposes of illustrative discussion of embodiments of the invention. In this regard, the description taken with the drawings makes apparent to those skilled in the art how embodiments of the invention may be practiced.

In the drawings:

[0052] FIG. 1A-1D is a graph with photos illustrating inhibition of metastatic disease by systemic administration of Ad5-PPE-1-3X fas-chimera adenovirus vector in the Lewis Lung Cancer model. FIGS. 1A-1C are photos of exemplary lungs from control (saline) and Ad5-PPE-1-3X fas-chimera adenovirus vector treated mice. Note the strong, dose dependent inhibition of metastatic development by the Ad5-PPE-1-3X fas-chimera adenovirus vector, as reflected in both the gross morphology and weight of the excised lungs (FIG. 1C);

[0053] FIG. 2 is a histogram showing the pre-treatment titer of pre-treatment neutralizing anti-Ad5 antibodies in the serum of patients in cohort 6 (3×10¹² vp), as a function of their disease progression at day 28 post administration of Ad5-PPE-1-3X fas-chimera adenovirus vector. Note the lack of correlation between progressive disease (blue), stable disease (red) and antibody titer (Y-axis);

[0054] FIGS. 3A and 3B are histograms showing the pre-treatment titer of neutralizing anti-Ad5 antibodies in the serum of patients, as a function of their disease progression at day 56 post administration of Ad5-PPE-1-3X fas-chimera adenovirus vector. FIG. 3A represents values for cohort 6. FIG. 3B represents the values for cohorts 6 and 7. Note the lack of correlation between progressive disease (blue), stable disease (red) and the baseline titer of neutralizing antibodies;

[0055] FIG. 4 is a baseline (pre-dose) cervical CT scan of a subject with advanced papillary thyroid cancer who received a single dose (3×10¹² virus particles) of Ad5-PPE-1-3X fas-chimera adenovirus vector, demonstrating a paratracheal metastasis causing partial obstruction of the trachea (red arrows);

[0056] FIG. 5 is a follow-up cervical CT of the same subject as described in FIG. 4, 6 months post-administration of Ad5-PPE-1-3X fas-chimera adenovirus vector, demonstrating regression of the metastatic lesion (red arrows) with central liquefaction (blue arrow);

[0057] FIGS. 6A-6C are abdominal CT scans illustrating regression of a metastatic lesion following administration of a single dose of Ad5-PPE-1-3X fas-chimera adenovirus vector (3×10¹² virus particles) in a subject with advanced neuroendocrine cancer. FIG. 6A is an abdominal CT scan showing the metastatic lesion (circled) in the liver at 21 days post-administration. FIG. 6B is an abdominal CT scan showing significant regression of the lesion (circled) in the liver at 50 days post-administration.

[0058] FIG. 6C is an abdominal CT scan showing even greater regression of the lesion (circled) in the liver at 112 days post-administration;

[0059] FIGS. 7A-7C are abdominal CT scans illustrating regression of a metastatic lesion following administration of a single dose of Ad5-PPE-1-3X fas-chimera adenovirus vector (3×10¹² virus particles) as described in FIGS. 6A-6C. FIGS. 7A-7C are CT scans from the same series, same subject as in FIGS. 6A-6C, showing the same lesion from a different orientation (CT slice at a different level);

[0060] FIG. 8 is a graph illustrating the disease response of individual patients in cohorts 1-5, measured at days 0, 28 and 56, and expressed as percent change in RECIST scores, relative to those observed on day 0. Greater increase in RECIST scores is typically indicative of progressive disease (solid line=stable disease, dotted line=progressive disease);

[0061] FIG. 9 is a graph illustrating the disease response of individual patients in cohorts 6 and 7, measured at days 0, 28 and 56, and expressed as percent change in RECIST scores, relative to those observed on day 0. Greater increase in RECIST scores is typically indicative of progressive disease (solid line=stable disease, dotted line=progressive disease). Note the trend to lesser change in RECIST scores among cohorts 6 and 7;

[0062] FIGS. 10A and 10B is a timeline showing the effects of administration of a single dose of Ad5-PPE-1-3X fas-chimera adenovirus vector (3×10¹² virus particles) on disease progression in two thyroid cancer patients (see Example II herein). FIG. 10A depicts the survival of a patient with refractory metastatic papillary thyroid cancer, receiving two doses the Ad5-PPE-1-3X fas-chimera adenovirus vector nearly two years apart, and has remained progression-free for most of that time. FIG. 10B shows the progression free survival of a patient with medullary thyroid cancer, who remained stable and progression free when monitored at 120 days after receiving a single dose of the Ad5-PPE-1-3X fas-chimera adenovirus vector. SD=stable disease. PD=progressive disease. PR=partial response;

[0063] FIG. 11 is a graph illustrating the humoral immune response to a single administration of the Ad5-PPE-1-3X fas-chimera adenovirus, in cohorts 1 to 7 (cohort 1-diamond .diamond-solid.; cohort 2=square .box-solid.; cohort 3=triangle .tangle-solidup.; cohort 4="X"; cohort 5=star; cohort 6=circle ; cohort 7=vertical line |). Individual patients' anti-Adenovirus 5 IgG levels were assayed (as detailed in Example II) at day 0 (administration), and days 7, 14, 21 and 28 post infusion. Note the plateau of the immune response as early as day 7 in some cohorts (e.g. 1, 3, 4) and later in others (e.g. cohorts 2, 5, 6, 7);

[0064] FIGS. 12A and 12B show the pharmacokinetics of the Ad5-PPE-1-3X fas-chimera adenovirus vector, following intravenous administration. FIG. 12A is a graph depicting the pharmacokinetics of the Ad5-PPE-1-3X fas-chimera adenovirus vector in blood, from administration to day 56, post-infusion. Whole blood samples of patients receiving 10¹⁰ (.diamond-solid., solid black line), 3×10¹⁹ (.box-solid., dotted black line), 10¹¹ (.tangle-solidup., dashed black line), 3×10¹¹ (X, solid grey line), 10¹² (.box-solid., dotted grey line) and 3×10¹² ( , dashed grey line) Ad5-PPE-1-3X fas-chimera adenovirus particles were analyzed by RT-PCR at indicated timepoints for adenovirus 5 DNA. Adenovirus levels were reduced by at least two orders of magnitude, or undetectable by day 56. FIG. 12B represents the average levels of adenovirus particles, analyzed by RT-PCR, in whole blood sampled at the end of the infusion with the virus vector;

[0065] FIGS. 13A and 13B illustrate the effects of combined Ad5-PPE-1-3X fas-chimera adenovirus vector and sunitinib on the Lewis Lung metastatic cancer model. Mice with induced lung metastases received either the Ad5-PPE-1-3X fas-chimera adenovirus vector or oral sunitinib, in the indicated dosages, or a combination of the two therapies. Control mice received empty (sham) virus vehicles. Lung metastases were evaluated according to the tumor mass in grams (tumor burden) at 22 days post-primary tumor removal. FIG. 13 A is a histogram of the values in FIG. 13B. Note the enhanced effect of the combination therapy at 80 mg/kg sunitinib, and 10⁹ Ad5-PPE-1-3X fas-chimera virus particles.

[0066] Unless otherwise defined, all technical and/or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the invention, exemplary methods and/or materials are described below. In case of conflict, the patent specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and are not intended to be necessarily limiting.

DESCRIPTION OF SPECIFIC EMBODIMENTS OF THE INVENTION

[0067] The present invention, in some embodiments thereof, relates to anti-angiogenic adenovirus vectors, and therapeutic use thereof, and more particularly, but not exclusively, to clinical protocols for treatment of solid tumors in patients with an Ad5-PPE-1-3X-fas-chimera adenovirus vector.

[0068] Before explaining at least one embodiment of the invention in detail, it is to be understood that the invention is not necessarily limited in its application to the details set forth in the following description or exemplified by the Examples. The invention is capable of other embodiments or of being practiced or carried out in various ways.

[0069] Angiogenesis is required for the development of neoplastic and hyperproliferative growths. Gene therapy for anti-angiogenic therapy in conditions associated with neovascularization, such as cancer, has been investigated, however, despite promising results in in-vitro experiments and in animal models, there has been little success with anti-angiogenic gene therapy in the clinical setting, likely due to obstacles including duration of expression of the transferred gene, induction of host immune response, cytotoxicity of the vectors and tissue specificity of expression.

[0070] The present inventors have developed a clinically safe and effective procedure for administration of a therapeutic recombinant adenovirus vector comprising a cytotoxic fas-chimera effector sequence under transcriptional control of an angiogenic endothelial-specific modified murine pre-pro endothelin promoter, which can be used for treatment of a variety of cancers and other hyperproliferative, neovascular-dependent diseases, for example, for solid tumors.

[0071] Thus, according to one aspect of the present invention, there is provided a method of treating a solid tumor in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a non-replicating adenovirus vector, said vector comprises a polynucleotide which comprises a fas-chimera transgene transcriptionally linked to a murine pre-proendothelin promoter, wherein said therapeutically effective amount is at least 1×10⁸ virus particles, thereby treating the solid tumor.

[0072] As used herein, the phrases "cancer", "malignancy", "solid tumor" or "hyperproliferative disorder" are used as synonymous terms and refer to any of a number of diseases that are characterized by uncontrolled, abnormal proliferation of cells, the ability of affected cells to spread locally or through the bloodstream and lymphatic system to other parts of the body (i.e., metastasize) as well as any of a number of characteristic structural and/or molecular features. A "cancerous" or "malignant cell" or "solid tumor cell" is understood as a cell having specific structural properties, lacking differentiation and being capable of invasion and metastasis. "Cancer" refers to all types of cancer or neoplasm or malignant tumors found in mammals, including carcinomas and sarcomas. Examples are cancers of the breast, lung, non-small cell lung, stomach, brain, head and neck, medulloblastoma, bone, liver, colon, genitourinary, bladder, urinary, kidney, testes, uterus, ovary, cervix, prostate, melanoma, mesothelioma, sarcoma, (see DeVita, et al., (eds.), 2001, Cancer Principles and Practice of Oncology, 6th. Ed., Lippincott Williams & Wilkins, Philadelphia, Pa.; this reference is herein incorporated by reference in its entirety for all purposes).

[0073] "Cancer-associated" refers to the relationship of a nucleic acid and its expression, or lack thereof, or a protein and its level or activity, or lack thereof, to the onset of malignancy in a subject cell. For example, cancer can be associated with expression of a particular gene that is not expressed, or is expressed at a lower level, in a normal healthy cell. Conversely, a cancer-associated gene can be one that is not expressed in a malignant cell (or in a cell undergoing transformation), or is expressed at a lower level in the malignant cell than it is expressed in a normal healthy cell.

[0074] "Hyperproliferative disease" refers to any disease or disorder in which the cells proliferate more rapidly than normal tissue growth. Thus, a hyperproliferating cell is a cell that is proliferating more rapidly than normal cells.

[0075] "Neovascularization" and "angiogenesis" refer to the growth of new blood vessels. Pathological angiogenesis or neovascularization refers to unbalanced new blood vessel growth, including non-self-limiting endothelial and periendothelial cell-proliferation. "Angiogenic diseases" are conditions of unregulated angiogenesis, for example, cancer, ocular neovascularization, arthritis, diabetes, skin diseases, chronic inflammatory diseases such as rheumatoid arthritis, psoriasis and synovitis.

[0076] "Advanced cancer" means cancer that is no longer localized to the primary tumor site, or a cancer that is Stage III or IV according to the American Joint Committee on Cancer (AJCC).

[0077] "Well tolerated" refers to the absence of adverse changes in health status that occur as a result of the treatment and would affect treatment decisions.

[0078] "Metastatic" refers to tumor cells, e.g., human solid tumor or thyroid malignancy, that are able to establish secondary tumor lesions in the lungs, liver, bone or brain of immune deficient mice upon injection into the mammary fat pad and/or the circulation of the immune deficient mouse.

[0079] A "solid tumor" includes, but is not limited to, sarcoma, melanoma, carcinoma, or other solid tumor cancer. "Sarcoma" refers to a tumor which is made up of a substance like the embryonic connective tissue and is generally composed of closely packed cells embedded in a fibrillar or homogeneous substance: Sarcomas include, but are not limited to, chondrosarcoma, fibrosarcoma, lymphosarcoma, melanosarcoma, myxosarcoma, osteosarcoma, Abemethy's sarcoma, adipose sarcoma, liposarcoma, alveolar soft part sarcoma, ameloblastic sarcoma, botryoid sarcoma, chloroma sarcoma, chorio carcinoma, embryonal sarcoma, Wilms' tumor sarcoma, endometrial sarcoma, stromal sarcoma, Ewing's sarcoma, fascial sarcoma, fibroblastic sarcoma, giant cell sarcoma, granulocytic sarcoma, Hodgkin's sarcoma, idiopathic multiple pigmented hemorrhagic sarcoma, immunoblastic sarcoma of B cells, lymphoma, immunoblastic sarcoma of T-cells, Jensen's sarcoma, Kaposi's sarcoma, Kupffer cell sarcoma, angiosarcoma, leukosarcoma, malignant mesenchymoma sarcoma, parosteal sarcoma, reticulocytic sarcoma, Rous sarcoma, serocystic sarcoma, synovial sarcoma, and telangiectaltic sarcoma.

[0080] "Melanoma" refers to a tumor arising from the melanocytic system of the skin and other organs. Melanomas include, for example, acral-lentiginous melanoma, amelanotic melanoma, benign juvenile melanoma, Cloudman's melanoma, S91 melanoma, Harding-Passey melanoma, juvenile melanoma, lentigo maligna melanoma, malignant melanoma, metastatic melanoma, nodular melanoma, subungal melanoma, and superficial spreading melanoma.

[0081] "Carcinoma" refers to a malignant new growth made up of epithelial cells tending to infiltrate the surrounding tissues and give rise to metastases. Exemplary carcinomas include, for example, acinar carcinoma, acinous carcinoma, adenocystic carcinoma, adenoid cystic carcinoma, carcinoma adenomatosum, carcinoma of adrenal cortex, alveolar carcinoma, alveolar cell carcinoma, basal cell carcinoma, carcinoma basocellulare, basaloid carcinoma, basosquamous cell carcinoma, bronchioalveolar carcinoma, bronchiolar carcinoma, bronchogenic carcinoma, cerebriform carcinoma, cholangiocellular carcinoma, chorionic carcinoma, colloid carcinoma, comedo carcinoma, corpus carcinoma, cribriform carcinoma, carcinoma en cuirasse, carcinoma cutaneum, cylindrical carcinoma, cylindrical cell carcinoma, duct carcinoma, carcinoma durum, embryonal carcinoma, encephaloid carcinoma, epiermoid carcinoma, carcinoma epitheliale adenoides, exophytic carcinoma, carcinoma ex ulcere, carcinoma fibrosum, gelatiniform carcinoma, gelatinous carcinoma, giant cell carcinoma, carcinoma gigantocellulare, glandular carcinoma, granulosa cell carcinoma, hair-matrix carcinoma, hematoid carcinoma, hepatocellular carcinoma, Hurthle cell carcinoma, hyaline carcinoma, hypemephroid carcinoma, infantile embryonal carcinoma, carcinoma in situ, intraepidermal carcinoma, intraepithelial carcinoma, Krompecher's carcinoma, Kulchitzky-cell carcinoma, large-cell carcinoma, lenticular carcinoma, carcinoma lenticulare, lipomatous carcinoma, lymphoepithelial carcinoma, carcinoma medullare, medullary carcinoma, melanotic carcinoma, carcinoma molle, mucinous carcinoma, carcinoma muciparum, carcinoma mucocellulare, mucoepidernoid carcinoma, carcinoma mucosum, mucous carcinoma, carcinoma myxomatodes, naspharyngeal carcinoma, oat cell carcinoma, carcinoma ossificans, osteoid carcinoma, papillary carcinoma, periportal carcinoma, preinvasive carcinoma, prickle cell carcinoma, pultaceous carcinoma, renal cell carcinoma of kidney, reserve cell carcinoma, carcinoma sarcomatodes, schneiderian carcinoma, scirrhous carcinoma, carcinoma scroti, signet-ring cell carcinoma, carcinoma simplex, small-cell carcinoma, solanoid carcinoma, spheroidal cell carcinoma, spindle cell carcinoma, carcinoma spongiosum, squamous carcinoma, squamous cell carcinoma, string carcinoma, carcinoma telangiectaticum, carcinoma telangiectodes, transitional cell carcinoma, carcinoma tuberosum, tuberous carcinoma, verrucous carcinoma, and carcinoma viflosum.

[0082] Additional cancers include, for example, Leukemia, Hodgkin's Disease, Non-Hodgkin's Lymphoma, multiple myeloma, neuroblastoma, breast cancer, ovarian cancer, lung cancer, rhabdomyosarcoma, primary thrombocytosis, primary macroglobulinemia, small-cell lung tumors, primary brain tumors, stomach cancer, colon cancer, malignant pancreatic insulanoma, malignant carcinoid, urinary bladder cancer, premalignant skin lesions, testicular cancer, lymphomas, thyroid cancer, papillary thyroid cancer, neuroblastoma, neuroendocrine cancer, esophageal cancer, genitourinary tract cancer, malignant hypercalcemia, cervical cancer, endometrial cancer, adrenal cortical cancer, and prostate cancer.

[0083] The present inventors have shown that systemic administration of the Ad5-PPE-1-3X-fas-c adenoviral vector was correlated with a reduction in tumor mass and prolonged stable disease in a patient suffering from metastatic papillary thyroid cancer (see Example II and FIGS. 4 and 5 that follow). Thus, according to one aspect of some embodiments of the present invention there is provided a method for treating a thyroid cancer in a patient in need thereof, the method comprising administering to the subject a single or multiple intravenous dose(s) of 3×10¹² or 10¹³ virus particles of a non-replicating adenovirus vector comprising a polynucleotide having a nucleotide sequence as set forth in SEQ ID NOs: 9 or 10. Disease progression in thyroid cancer can be assessed or monitored by methods including, but not limited to, radiographic analysis of tumor mass and density (e.g. RECIST criteria), measurement of thyroid and thyroid-associated hormone levels, thyroglobulin levels, and the like.

[0084] The present inventors have shown that systemic administration of the Ad5-PPE-1-3X-fas-c adenoviral vector was correlated with a reduction in a metastatic hepatic lesion and prolonged stable disease in a patient suffering from metastatic neuroendocrine cancer (see Example II and FIGS. 6A-6C, 7A-7C that follow). Thus, according to one aspect of some embodiments of the present invention there is provided a method for treating a neuroendocrine cancer in a patient in need thereof, the method comprising administering to the subject a single or multiple intravenous dose(s) of 3×10¹² or 10¹³ virus particles of a non-replicating adenovirus vector comprising a polynucleotide having a nucleotide sequence as set forth in SEQ ID NOs: 9 or 10. Methods for assessing or monitoring disease progression in neuroendocrine cancer include radiographic analysis of tumor mass and density (e.g. RECIST criteria), measurement of liver enzyme levels (where the tumor is a liver metastasis), and the like.

[0085] The method of the present invention as claimed has been indicated effective in treating other cancers. Thus, according to further aspects of some embodiments of the present invention there is provided a method for treating an ovarian cancer, a non small cell lung cancer and/or a renal cell carcinoma in a patient in need thereof, the method comprising administering to the subject a single or multiple intravenous dose(s) of 3×10¹² or 10¹³ virus particles of a non-replicating adenovirus vector comprising a polynucleotide having a nucleotide sequence as set forth in SEQ ID NOs: 9 or 10. Methods for assessing or monitoring disease progression in these cancers include radiographic analysis of tumor mass and density (e.g. RECIST criteria), measurement of organ function (e.g. kidney function in renal cell carcinoma), measurement of specific biomarkers, endocrine function, and the like.

[0086] Contemplated subjects to be treated include mammals e.g. humans. According to one embodiment the subject has received a prior treatment for the solid tumor (e.g. radiotherapy and/or chemotherapy) and the malignant tumor has relapsed. According to another embodiment, the subject has not received a prior treatment for the malignant tumor.

[0087] The phrase "viral vector" refers to a replication-competent or replication-deficient viral particle which is capable of transferring nucleic acid molecules into a host.

[0088] The present inventors contemplate use of Replication Defective Vectors and Replication Defective Vector-Producing Packaging Cells. Examples of such vectors are adenoviral vectors, AAV vectors and retroviral vectors and others described in Shir et al, Cellular and Molecular Neurobiology, Vol. 21, No. 6, December 2001, the contents of which are incorporated herein by reference.

[0089] The term "virus" refers to any of the obligate intracellular parasites having no protein-synthesizing or energy-generating mechanism. The viral genome may be RNA or DNA contained with a coated structure of protein of a lipid membrane. Examples of viruses useful in the practice of the present invention include baculoviridiae, parvoviridiae, picornoviridiae, herepesviridiae, poxyiridiae, adenoviridiae, picotrnaviridiae. The term recombinant virus includes chimeric (or even multimeric) viruses, i.e. vectors constructed using complementary coding sequences from more than one viral subtype. (See, e.g. Feng, et al. Nature Biotechnology 15:866-870) The term "adenovirus" is synonymous with the term "adenoviral vector" and refers to viruses of the genus adenoviridiae. The term adenoviridiae refers collectively to animal adenoviruses of the genus mastadenovirus including but no limited to human, bovine, ovine, equine, canine, porcine, murine and simian adenovirus subgenera. In particular, human adenoviruses includes the A-F subgenera as well as the individual serotypes thereof the individual serotypes and A-F subgenera including but not limited to human adenovirus types 1, 2, 3, 4, 4a, 5, 6, 7, 8, 9, 10, 11 (Ad11A and Ad 11P), 12, 13, 14, 15, 16, 17, 18, 19, 19a, 20, 21, 22, 23, 24, 2.5, 26, 27, 28, 29, 30, 31, 32, 33, 34, 34a, 35, 35p, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, and 91. The term bovine adenoviruses includes but is not limited to bovine adenovirus types 1, 2, 3, 4, 7, and 10. The term canine adenoviruses includes but is not limited to canine types 1 (strains CLL, Glaxo, RI261, Utrect, Toronto 26-61) and 2. The term equine adenoviruses includes but is not limited to equine types 1 and 2. The term porcine adenoviruses includes but is not limited to porcine types 3 and 4. In one embodiment of the invention, the adenovirus is derived from the human adenovirus serotypes 2 or 5. For purposes of this invention, adenovirus vectors can be replication-competent or replication deficient in a target cell. In some embodiments, the adenovirus vectors are conditionally or selectively replicating adenoviruses, wherein a gene[s] required for viral replication is [are] operatively linked to a cell and/or context-specific promoter. Examples of selectively replicating or conditionally replicating viral vectors are known in the art (see, for example, U.S. Pat. No. 7,691,370). In one embodiment, the adenovirus vector is a conditionally replicating adenovirus wherein the E1 gene is under transcriptional control of the pre-proendothelin promoter PPE-1 (PPE-1, SEQ ID NO: 13). In another embodiment, the adenovirus vector is a conditionally replicating or selectively replicating adenovirus wherein the E1 gene is under transcriptional control of the modified pre-proendothelin promoter PPE-1-3X (PPE-1-3X, SEQ ID NO: 12). In some embodiments, adenovirus vectors suitable for use with the present invention include all adenovirus serotypes having hexon protein structure. Viral vectors suitable for therapeutic use include adenoviral vectors, retrovirusal vectors, AAV, herpesvirus vectors and the like. Engineering and production of viral vectors is well known in the art, as described in detail in, for example, U.S. Pat. No. 7,732,129 or 6,649,158, which are incorporated herein by reference, in their entirety. In specific embodiments, the adenovirus is a C-type adenovirus (Ad5, Ad2), a B-type adenovirus (Ad3, Ad16, Ad21, Ad35, Ad50), an E-type adenovirus (Ad4) or an F-type adenovirus (Ad41).

[0090] As used herein, the phrase adenoviral vector refers to a vector in which, among the nucleic acid molecules in the viral particle, sequences necessary to function as a viral vector are based on the adenoviral genome.

[0091] According to one embodiment the adenoviral vector is a non-replicating serotype 5 (Ad5) adenoviral vector.

[0092] According to another embodiment, the adenoviral vector comprises a sequence as set forth in SEQ ID NO: 1 or SEQ ID NO: 11.

[0093] It will be appreciated that the present invention also contemplates use of oncolytic viruses which reproduce themselves in cancer cells and subsequently kill the initially infected cells by lysis. Such viruses proceed to infect adjacent cells thus repeating the cycle. Contemplated examples of oncolytic viruses include, but are not limited to Herpes Simplex Virus, conditionally replicative Ads (CRAds) and reoviruses.

[0094] Two major strategies for development of CRAd vectors have been developed, mainly focusing on the genetic engineering of the early 1 (E1) genes to restrict virus replication to target cells and to spare normal tissue. Genetic complementation-type (type 1) CRAds, such as Ad524, have a mutation in the immediately early (E1A) or early (E1B) adenoviral region, which is complemented in tumor cells but not in normal cells. In trans complementation-type (type 2) CRAds, virus replication is controlled via a tumor/tissue-specific promoter.

[0095] Reovirus is a naturally occurring oncolytic virus that requires activated Ras signaling pathways of tumor cells for its replication. Ras pathways are activated in most malignant tumors via upstream signaling by receptor tyrosine kinases.

[0096] As mentioned the viral vectors of this aspect of the present invention comprise a cytotoxic fas-chimera effector sequence under transcriptional control of an angiogenic endothelial-specific modified murine pre-pro endothelin promoter.

[0097] Typically, such viral vectors are constructed using genetic recombination technology--i.e. recombinant viral vectors.

[0098] The Fas-chimera (Fas-c) polypeptide, is a previously described fusion of two "death receptors", constructed from the extracellular region of TNFR1 (SEQ ID NO: 2) and the trans-membrane and intracellular regions of Fas (SEQ ID NO: 3) [Boldin M P et al. J Biol Chem (1995) 270(14):7795-8; the contents of which are incorporated herein by reference].

[0099] According to one embodiment the Fas-c is encoded by a polynucleotide as set forth in SEQ ID NO: 4.

[0100] It will be appreciated that the present invention also contemplates use of a viral construct (e.g. an adenoviral construct) comprising an endothelial/periendothelial cell-specific promoter operatively linked to other cytotoxic polypeptides for the treatment of solid tumors.

[0101] Such polypeptides, include but are not limited to suicide polypeptides such as p53 and egr-1-TNF-alpha, cytotoxic pro-drug/enzymes for drug susceptibility therapy such as ganciclovir/thymidine kinase and 5-fluorocytosine/cytosine deaminase, and antimetastatic polypeptides such as 5 E1A.

[0102] The term "promoter" as used herein refers to a DNA sequence which directs transcription of a polynucleotide sequence operatively linked thereto in the cell in a constitutive or inducible manner. The promoter may also comprise enhancer elements which stimulate transcription from the linked promoter.

[0103] The pre-pro endothelial promoter as used herein refers to the preproendothelin-1 (PPE-1) promoter, of mammalian origin. In one embodiment, the pre-proendothelin 1 promoter is a murine pre-pro endothelin 1 promoter (PPE-1, SEQ ID NO: 13) and modifications thereof. It will be appreciated that other endothelial specific promoters can be used with the present invention, for example, the TIE-1 promoter, the TIE-2 promoter, the Endoglin promoter, the von Willerband promoter, the ICDR/flk-1 promoter, The FLT-1 promoter, the Egr-1 promoter, the ICAM-1 promoter, the VCAM-1 promoter, the PECAM-1 promoter and the aortic carboxypeptidase-like protein (ACLP) promoter.

[0104] According to one embodiment the promoter comprises at least one copy of an enhancer element that confers endothelial cell specific transcriptional activity. According to one embodiment the enhancer element is naturally found positioned between the -364 bp and -320 bp of the murine PPE-1 promoter (as set forth in SEQ ID NO: 6). In one embodiment, the promoter comprises at least two and more preferably three of the above described enhancer elements. According to a specific embodiment, the promoter comprises two of the above described enhancer elements on one strand of the promoter DNA and one of the above described enhancer element on the complementary strand of the promoter DNA.

[0105] In yet another embodiment, the promoter comprises a modified enhancer element as set forth in SEQ ID NO: 8, optionally in combination with other enhancer elements. Thus, according to this embodiment, the promoter comprises a sequence as set forth in SEQ ID NO: 7.

[0106] According to another embodiment, the promoter further comprises at least one hypoxia response element--e.g. comprising a sequence as set forth in SEQ ID NO: 5.

[0107] An exemplary promoter which can be used in the context of the present invention comprises a sequence as set forth in SEQ ID NO: 12. This sequence comprises SEQ ID NO: 5 and SEQ ID NO: 7 (which itself comprises two copies of SEQ ID NO: 6 either side of one copy of SEQ ID NO: 8).

[0108] According to a particular embodiment of this aspect of the present invention, the viral vector consists of a sequence as set forth in SEQ ID NOs: 9 or 10.

[0109] The Ad5-PPE-1-3X-fas-c sequence, as set forth in SEQ ID NO: 9 or 10 comprises a sequence which is an anti-sense copy of SEQ ID NO: 7, located at nucleic acid coordinates 894-1036, a sequence which is a single antisense copy of SEQ ID NO: 8 located at nucleotide coordinates 951-997; a sequence which is a first antisense copy of SEQ ID NO: 6 located at nucleotide coordinates 907-950; a sequence which is a second antisense copy of SEQ ID NO: 6 located at nucleotide coordinates 993-1036; and a third copy of SEQ ID NO: 6 in the sense orientation at position 823-866.

[0110] In some embodiments of the invention, the viral vector comprises additional polynucleotide sequences capable of enhancing or inhibiting transcriptional activity of an endothelial specific promoter. According to an aspect of some embodiments of the invention, the additional polynucleotide sequence includes an isolated polynucleotide comprising at least 6 nucleotides of element X of a pre-proendothelin (PPE-1) promoter, the element X having a wild type sequence as set forth by SEQ ID NO:6, wherein the at least 6 nucleotides comprise at least 2 consecutive sequences derived from SEQ ID NO:6, each of the at least 2 consecutive sequences comprises at least 3 nucleotides, at least one of the at least 3 nucleotide being positioned next to at least one nucleotide position in SEQ ID NO:6, the at least one nucleotide position in SEQ ID NO:6 is selected from the group consisting of:

[0111] (i) at least one nucleotide of wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC);

[0112] (ii) at least one nucleotide of wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG);

[0113] (iii) at least one nucleotide of wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC);

[0114] (iv) at least one nucleotide of wild type M6 sequence set forth by SEQ ID NO: 17 (GGGTG);

[0115] (v) at least one nucleotide of wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT);

[0116] (vi) at least one nucleotide of wild type M1 sequence set forth by SEQ ID NO: 20 (GTACT); and

[0117] (v) at least one nucleotide of wild type M3 sequence set forth by SEQ JD NO: 21 (CTTTT);

[0118] wherein the at least one nucleotide position is mutated as compared to SEQ ID NO:6 by at least one nucleotide substitution, at least one nucleotide deletion and/or at least one nucleotide insertion, with the proviso that a mutation of the at least one nucleotide position does not result in nucleotides GGTA at position 21-24 of SEQ ID NO:6 and/or in nucleotides CATG at position 29-32 of SEQ ID NO:6, such that when the isolated polynucleotide is integrated into the PPE-1 promoter and placed upstream of a reporter gene (e.g., luciferase coding sequence) the expression level of the reporter gene is upregulated or downregulated as compared to when SEQ ID NO:6 is similarly integrated into the PPE-1 promoter and placed upstream of the reporter gene coding sequence.

[0119] According to some embodiments of the invention, the isolated polynucleotide is not naturally occurring in a genome or a whole chromosome sequence of an organism.

[0120] As used herein the phrase "naturally occurring" refers to as found in nature, without any man-made modifications.

[0121] As described above, the at least 6 nucleotides of element X comprise at least 2 consecutive sequences derived from SEQ ID NO:6.

[0122] As used herein the phrase "consecutive sequence derived from SEQ ID NO:6" refers to a nucleic acid sequence (a polynucleotide) in which the nucleotides appear in the same order as in the nucleic acid sequence of SEQ ID NO:6 from which they are derived. It should be noted that the order of nucleotides is determined by the chemical bond (phosphodiester bond) formed between a 3'-OH of a preceding nucleotide and the 5'-phosphate of the following nucleotide.

[0123] According to some embodiments of the invention, each of the at least 2 consecutive sequences comprises at least 3 nucleotides, e.g., 3 nucleotides, 4 nucleotides, 5 nucleotides, 6 nucleotides, 7 nucleotides, 8 nucleotides, 9 nucleotides, 10 nucleotides, 11 nucleotides, 12 nucleotides, 13 nucleotides, 14 nucleotides, 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, 28 nucleotides, 29 nucleotides, 30 nucleotide, 31 nucleotides, 32 nucleotides, 33 nucleotides, 34 nucleotides, 35 nucleotides, 36 nucleotides, 37 nucleotides, 38 nucleotides, 39 nucleotides, 40 nucleotides, 41 nucleotides of SEQ ID NO:6.

[0124] As described, the isolated polynucleotide comprises at least 2 consecutive sequences derived from SEQ ID NO:6. According to some embodiments of the invention, the isolated polynucleotide comprises 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 consecutive sequences derived from SEQ ID NO:6.

[0125] As used herein the phrase "wild type" with respect to a nucleotide sequence refers to the nucleic acid sequence as appears in SEQ ID NO:6. Examples include, but are not limited to wild type M4 sequence (SEQ ID NO: 15), wild type M5 sequence (SEQ ID NO: 16), wild type M8 (SEQ ID NO:19), wild type M6 sequence (SEQ ID NO:17), wild type M7 sequence (SEQ ID NO:18), wild type M1 (SEQ ID NO:20) and wild type M3 sequence (SEQ ID NO:21).

[0126] According to some embodiments of the invention, the mutation is an insertion of at least one nucleotide in a nucleotide position with respect to SEQ ID NO:6. According to some embodiments of the invention, the insertion includes at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 nucleotides, e.g., at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 55, at least about 60, at least about 65, at least about 70, at least about 75, at least about 80, at least about 85, at least about 90, at least about 95, at least about 100, at least about 200, at least about 300, or more nucleotides.

[0127] It should be noted that the sequence which is inserted by the mutation can be derived from any source (e.g., species, tissue or cell type), and is not limited to the source of the sequence of element X.

[0128] According to some embodiments of the invention, the mutation is a combination of any of the mutation types described above, i.e., substitution, insertion and deletion. For example, while one nucleotide position in SEQ ID NO:6 can be subject to a substitution mutation, another nucleotide position in SEQ ID NO:6 can be subject to a deletion or insertion. Additionally or alternatively, while one nucleotide position in SEQ ID NO:6 can be subject to a deletion mutation, another nucleotide position in SEQ ID NO:6 can be subject to a substitution or insertion. Additionally or alternatively, while one nucleotide position in SEQ ID NO:6 can be subject to an insertion mutation, another nucleotide position in SEQ ID NO:6 can be subject to a substitution or deletion. It should be noted that various other combinations are possible.

[0129] According to specific embodiments of the invention, the mutation in the isolated polynucleotide of the invention does not result in nucleotides GGTA at position 21-24 of SEQ ID NO:6 and/or in nucleotides CATG at position 29-32 of SEQ ID NO:6.

[0130] As used herein the phrase "integrated into the PPE-1 promoter" refers to a nucleotide sequence (the isolated polynucleotide) which is covalently conjugated within the PPE-1 promoter sequence.

[0131] According to some embodiments of the invention, the isolated polynucleotide further comprises at least one copy of a nucleic acid sequence selected from the group consisting of:

[0132] (i) wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC),

[0133] (ii) wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG),

[0134] (iii) wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC),

[0135] (iv) wild type M6 sequence set forth by SEQ ID NO: 17 (GGGTG),

[0136] (v) wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT);

[0137] (vi) wild type M1 sequence set forth by SEQ ID NO: 20 (GTACT), and

[0138] (vii) wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT).

[0139] According to some embodiments of the invention, the isolated polynucleotide is integrated into (within), downstream of, or upstream of any known (or unknown) promoter sequence to thereby regulate (e.g., increase, decrease, modulate tissue-specificity, modulate inductive or constitutive expression) the transcriptional promoting activity of the promoter.

[0140] According to some embodiments of the invention, the isolated polynucleotide is for increasing expression of a heterologous polynucleotide operably linked thereto in endothelial cells. Such a polynucleotide can include wild type sequences of M4 and/or M5 in the presence or absence of additional sequences from element X, and/or in the presence of other mutated sequences from element X.

[0141] According to some embodiments of the invention, the isolated polynucleotide comprises at least one copy of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC).

[0142] According to some embodiments of the invention, the isolated polynucleotide comprises at least one copy of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG).

[0143] According to some embodiments of the invention, the isolated polynucleotide comprises at least one copy of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC) and at least one copy of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG).

[0144] According to some embodiments of the invention, the at least one nucleotide position which is mutated as compared to SEQ ID NO:6 is at least one nucleotide of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC). It should be noted that such an isolated polynucleotide may further include a wild type M6 sequence (SEQ ID NO:17) and/or a wild type M7 sequence (SEQ ID NO:18)

[0145] Non-limiting examples of isolated polynucleotides which include at least one copy of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC) and a mutation in at least one nucleotide of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs:55-62.

[0146] Non-limiting examples of isolated polynucleotides which include at least one copy of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG) and a mutation in at least one nucleotide of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs: 63-66.

[0147] Non-limiting examples of isolated polynucleotides which include at least one copy of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), at least one copy of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG) and a mutation in at least one nucleotide of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs: 67-70.

[0148] According to some embodiments of the invention, the isolated polynucleotide further comprising at least one copy of wild type M1 sequence set forth by SEQ ID NO: 20 (GTACT).

[0149] Non-limiting examples of isolated polynucleotides which include at least one copy of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), at least one copy of the wild type M1 sequence set forth by SEQ ID NO: 20 (GTACT), and a mutation in at least one nucleotide of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs: 71-105.

[0150] Non-limiting examples of isolated polynucleotides which include at least one copy of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M1 sequence set forth by SEQ ID NO: 20 (GTACT) and a mutation in at least one nucleotide of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs: 106-136.

[0151] Non-limiting examples of isolated polynucleotides which include at least one copy of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), at least one copy of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M1 sequence set forth by SEQ ID NO: 20 (GTACT) and a mutation in at least one nucleotide of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs:137-152.

[0152] According to some embodiments of the invention, the isolated polynucleotide reduces expression of a heterologous polynucleotide operably linked thereto in endothelial cells. Such a polynucleotide can include mutations in M4 and/or M5 in the presence or absence of additional sequences from element X, and/or in the presence of other mutated sequences from element X.

[0153] According to some embodiments of the invention, the at least one nucleotide position which is mutated as compared to SEQ ID NO:6 is at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC).

[0154] Non-limiting examples of isolated polynucleotides which includes a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO:46 (CATTC) are provided in SEQ ID NOs:153-162.

[0155] According to some embodiments of the invention, the at least one nucleotide position which is mutated as compared to SEQ ID NO:6 is at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG).

[0156] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG) are provided in SEQ ID NOs:163-171.

[0157] According to some embodiments of the invention, the at least one nucleotide position which is mutated as compared to SEQ ID NO:6 is at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC) and at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG).

[0158] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC) and a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG) are provided in SEQ ID NOs:172-180.

[0159] According to some embodiments of the invention, the isolated polynucleotide is for increasing expression of a heterologous polynucleotide operably linked thereto in cells other than endothelial cells. Such a polynucleotide can include mutations in M4 and/or M5 and wild type sequences of M6 and/or M7, in the presence or absence of additional sequences from element X, and/or in the presence of other mutated sequences from element X.

[0160] According to some embodiments of the invention, the isolated polynucleotide comprises a mutation in M4 (SEQ ID NO: 15) and/or in M5 (SEQ ID NO: 16) and at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and/or at least one copy of wild type M7 set forth by SEQ ID NO:18.

[0161] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC) and at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) are provided in SEQ ID NOs:181-182.

[0162] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG) and at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) are provided in SEQ ID NOs:183-189.

[0163] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG) and at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) are provided in SEQ ID NOs:190-191.

[0164] According to some embodiments of the invention, the isolated polynucleotide further comprises at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT).

[0165] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC) and at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT) are provided in SEQ ID NOs:192-195.

[0166] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG) and at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT) are provided in SEQ ID NOs:196-198.

[0167] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG) and at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT) are provided in SEQ ID NOs:199-202.

[0168] According to some embodiments of the invention, the isolated polynucleotide further comprises at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT).

[0169] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT) are provided in SEQ ID NOs:203-205.

[0170] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT) are provided in SEQ ID NOs:206-207.

[0171] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT) are provided in SEQ ID NOs:208-209.

[0172] According to some embodiments of the invention, the isolated polynucleotide reduces expression in cells of a heterologous polynucleotide operably linked thereto. Such a polynucleotide can include mutations in M4, M5, M6 and/or M7, in the presence or absence of additional sequences from element X, and/or in the presence of other mutated sequences from element X.

[0173] According to some embodiments of the invention, the isolated polynucleotide comprises at least one mutation in wild type M4 (SEQ ID NO: 15) and/or in wild type M5 (SEQ ID NO:47) and in wild type M6 set forth by SEQ ID NO: 17 (GGGTG).

[0174] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC) and a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) are provided in SEQ ID NOs:210-213.

[0175] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG) and a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) are provided in SEQ ID NOs:214-222.

[0176] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), and a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) are provided in SEQ ID NOs:223-231.

[0177] According to some embodiments of the invention, the isolated polynucleotide further comprises at least one mutation in wild type M7 set forth by SEQ ID NO: 18 (ACTTT).

[0178] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC) and a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT) are provided in SEQ ID NOs:232-236.

[0179] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG) and a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT) are provided in SEQ ID NOs:237-240.

[0180] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), and a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT) are provided in SEQ ID NOs:241-248.

[0181] According to some embodiments of the invention, the isolated polynucleotide further comprises at least one mutation in wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and at least one mutation in wild type M7 set forth by SEQ ID NO: 18 (ACTTT).

[0182] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT) are provided in SEQ ID NOs:249-258.

[0183] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT) are provided in SEQ ID NOs:259-264.

[0184] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT) are provided in SEQ ID NOs:265-270.

[0185] According to some embodiments of the invention, the isolated polynucleotide comprises at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) with additional wild type or mutated sequences derived from element X (SEQ ID NO:6).

[0186] Non-limiting examples of isolated polynucleotides which includes a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC) and at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs:271-279.

[0187] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG) and at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs:280-287.

[0188] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG) and at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs:288-291. Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs:294-298.

[0189] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and at least one copy of the wild type M8 sequence set forth by SEQ 11) NO: 19 (GCTTC) are provided in SEQ ID NOs:299-301.

[0190] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs:302-303.

[0191] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs:304-308.

[0192] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs:309-311.

[0193] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs:312-315.

[0194] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NO:316.

[0195] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NO:317.

[0196] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NO:318.

[0197] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs:319-327.

[0198] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs:328-333.

[0199] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs:334-337.

[0200] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs:338-344.

[0201] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs:345-348.

[0202] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs:349-354.

[0203] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs:355-361.

[0204] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs:362-365.

[0205] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) are provided in SEQ ID NOs:366-369.

[0206] According to some embodiments of the invention, the isolated polynucleotide comprises at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) with additional wild type or mutated sequences derived from element X (SEQ ID NO:6).

[0207] Non-limiting examples of isolated polynucleotides which includes a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:378-384.

[0208] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:628-634.

[0209] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:370-377.

[0210] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:385-390.

[0211] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:391-396.

[0212] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:397-401.

[0213] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:402-409.

[0214] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:410-417.

[0215] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:418-423.

[0216] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:424-425.

[0217] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACM) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:538-540.

[0218] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NO:426.

[0219] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:427-435.

[0220] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:436-444.

[0221] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:445-451.

[0222] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:452-458.

[0223] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:459-465.

[0224] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NO:466.

[0225] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:467-471.

[0226] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:472-477.

[0227] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:478-483.

[0228] According to some embodiments of the invention, the isolated polynucleotide further comprises at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) with additional wild type or mutated sequences derived from element X (SEQ ID NO:6).

[0229] Non-limiting examples of isolated polynucleotides which includes a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:484-495.

[0230] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:496-507.

[0231] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:508-515.

[0232] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:516-519.

[0233] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:520-523.

[0234] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC), and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:524-525.

[0235] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT), at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:526-529.

[0236] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT), at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:530-533.

[0237] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT), at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:534-535.

[0238] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT), at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:536-537.

[0239] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT) at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:538-539.

[0240] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), at least one copy of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), at least one copy of the wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT), at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NO:540.

[0241] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:541-547.

[0242] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:548-554.

[0243] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:555-559.

[0244] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT), at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:560-566.

[0245] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT), at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:567-573.

[0246] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT), at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:574-578.

[0247] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT), at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (curio are provided in SEQ ID NOs:579-583.

[0248] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT), at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:584-588.

[0249] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of the wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC), a mutation in at least one nucleotide of the wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG), a mutation in at least one nucleotide position of the wild type M6 set forth by SEQ ID NO: 17 (GGGTG), a mutation in at least one nucleotide position of the wild type M7 set forth by SEQ ID NO: 18 (ACTTT), at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) are provided in SEQ ID NOs:589-592.

[0250] According to some embodiments of the invention, the isolated polynucleotide comprises at least one-copy of wild type M3 sequence (SEQ ID NO: 21) and at least one copy of wild type M8 sequence (SEQ ID NO: 19), with at least one mutation in wild type M6 (SEQ ID NO: 17) and/or in wild type M7 (SEQ ID NO:50).

[0251] Non-limiting examples of isolated polynucleotides which include at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT), with a mutation in at least one nucleotide of the wild type M6 sequence (SEQ ID NO: 17), and/or a mutation in at least one nucleotide of the wild type M7 (SEQ ID NO: 18) are provided in SEQ ID NOs:593-600.

[0252] The present inventors have envisaged that an isolated polynucleotide which includes the wild type M8 sequence (SEQ ID NO: 19) and/or the wild type M3 (SEQ ID NO: 21) sequence in addition to tissue specific enhancers (e.g., wild type M4 and/or wild type M5), and/or induced enhancers (e.g., developmentally related- or stress related-enhancers) is expected to exert a more specific regulatory effect by suppressing expression in non-target cells or under non-induced conditions.

[0253] According to some embodiments of the invention, the isolated polynucleotide comprises at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and an endothelial specific enhancer sequence.

[0254] According to some embodiments of the invention, the isolated polynucleotide comprises at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of wild type M4 sequence set forth by SEQ ID NO: 15.

[0255] According to some embodiments of the invention, the isolated polynucleotide comprises at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of wild type M5 sequence set forth by SEQ ID NO:16.

[0256] According to some embodiments of the invention, the isolated polynucleotide comprises at least one copy of the wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC), at least one copy of wild type M4 sequence set forth by SEQ ID NO: 15 and at least one copy of wild type M5 sequence set forth by SEQ ID NO:16.

[0257] According to some embodiments of the invention, the isolated polynucleotide comprises at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) and an endothelial specific enhancer sequence.

[0258] According to some embodiments of the invention, the isolated polynucleotide comprises at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) and at least one copy of wild type M4 sequence set forth by SEQ ID NO: 15.

[0259] According to some embodiments of the invention, the isolated polynucleotide comprises at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT) and at least one copy of wild type M5 sequence set forth by SEQ ID NO:16.

[0260] According to some embodiments of the invention, the isolated polynucleotide comprises at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT), at least one copy of wild type M4 sequence set forth by SEQ ID NO: 15 and at least one copy of wild type M5 sequence set forth by SEQ ID NO:16.

[0261] According to some embodiments of the invention, the isolated polynucleotide comprises at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT), at least one copy of wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and an endothelial specific enhancer sequence.

[0262] According to some embodiments of the invention, the isolated polynucleotide comprises at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT), at least one copy of wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of wild type M4 sequence set forth by SEQ ID NO: 15.

[0263] According to some embodiments of the invention, the isolated polynucleotide comprises at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT), at least one copy of wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one copy of wild type M5 sequence set forth by SEQ ID NO: 16.

[0264] According to some embodiments of the invention, the isolated polynucleotide comprises at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT), at least one copy of wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC), at least one copy of wild type M4 sequence set forth by SEQ ID NO: 15 and at least one copy of wild type M5 sequence set forth by SEQ ID NO: 16.

[0265] According to some embodiments of the invention, the isolated polynucleotide comprises at least one copy of the wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT), at least one copy of wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC) and at least one enhancer element such as wild type M6 (SEQ ID NO: 17) and/or wild type M7 sequence (SEQ ID NO:18).

[0266] According to some embodiments of the invention, the isolated polynucleotide includes at least one copy of wild type M8 with additional flanking sequences such as at least one copy of a wild type M8 sequence (SEQ ID NO:19), at least one copy of wild type M7 (SEQ ID NO: 18) and/or wild type M9 sequence (SEQ ID NO: 14, CTGGA); and/or the isolated polynucleotide includes at least one copy of wild type M8 and at least one mutation in M7, with or without M9 (SEQ ID NO: 22). Such polynucleotides can be used as a non-specific repressor.

[0267] According to some embodiments of the invention, the isolated polynucleotide is for increasing expression of a heterologous polynucleotide operably linked thereto in cells/tissues.

[0268] According to some embodiments of the invention, the isolated polynucleotide comprises at least one copy of wild type M6 sequence set forth by SEQ ID NO: 17 (GGGTG) and/or at least one copy of wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT).

[0269] According to some embodiments of the invention, the isolated polynucleotide includes at least one copy of wild type M6 (SEQ ID NO: 17) and a mutation in at least one nucleotide of wild type M8 (SEQ ID NO: 19).

[0270] Non-limiting examples of isolated polynucleotide which include at least one copy of wild type M6 (SEQ ID NO: 17) and a mutation in at least one nucleotide of the wild type M8 (SEQ ID NO: 19) are provided in SEQ ID NOs:23-26.

[0271] According to some embodiments of the invention, the isolated polynucleotide includes at least one copy of wild type M7 (SEQ ID NO: 18) and a mutation in at least one nucleotide of wild type M8 (SEQ ID NO: 19).

[0272] Non-limiting examples of isolated polynucleotide which include at least one copy of wild type M7 (SEQ ID NO: 18) and a mutation in at least one nucleotide of the wild type M8 (SEQ ID NO: 19) are provided in SEQ ID NOs:27-28.

[0273] According to some embodiments of the invention, the isolated polynucleotide includes at least one copy of wild type M6 (SEQ ID NO: 17), at least one copy of wild type M7 (SEQ ID NO: 18) and a mutation in at least one nucleotide of wild type M8 (SEQ ID NO: 19).

[0274] According to some embodiments of the invention, the isolated polynucleotide includes at least one copy of wild type M1 (SEQ ID NO: 20) and a mutation in at least one nucleotide of wild type M8 (SEQ ID NO: 19).

[0275] Non-limiting examples of isolated polynucleotide which include at least one copy of wild type M1 (SEQ ID NO: 20) and a mutation in at least one nucleotide of the wild type M8 (SEQ ID NO: 19) are provided in SEQ ID NOs:43-54 and 601-632.

[0276] According to some embodiments of the invention, the isolated polynucleotide includes at least one copy of wild type M1 (SEQ ID NO: 20), at least one copy of wild type M6 (SEQ ID NO: 17) and/or at least one copy of wild type M7 (SEQ ID NO: 18) and a mutation in at least one nucleotide of wild type M8 (SEQ ID NO: 19).

[0277] Non-limiting examples of isolated polynucleotides which include a mutation in at least one nucleotide of wild type M8 (SEQ ID NO: 19) and at least one copy of wild type M1 (SEQ ID NO: 20), wild type M6 (SEQ ID NO: 17) and/or wild type M7 (SEQ ID NO: 18) are provided in SEQ ID NOs:29-42.

[0278] Additional examples of regulatory isolated polynucleotides which can be used according to some embodiments of the invention are provided (; SEQ ID NOs: 633-644) in the Examples section which follows.

[0279] According to an aspect of some embodiments of the invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence which comprises a first polynucleotide comprising the pre-proendothelin (PPE-1) promoter set forth by SEQ ID NO:13 and a second polynucleotide comprising at least one copy of a nucleic acid sequence selected from the group consisting of:

[0280] (i) wild type M4 sequence set forth by SEQ ID NO: 15 (CATTC),

[0281] (ii) wild type M5 sequence set forth by SEQ ID NO: 16 (CAATG),

[0282] (iii) wild type M8 sequence set forth by SEQ ID NO: 19 (GCTTC),

[0283] (iv) wild type M6 sequence set forth by SEQ ID NO: 17 (GGGTG),

[0284] (v) wild type M7 sequence set forth by SEQ ID NO: 18 (ACTTT);

[0285] (vi) wild type M1 sequence set forth by SEQ ID NO: 20 (GTACT), and

[0286] (vii) wild type M3 sequence set forth by SEQ ID NO: 21 (CTTTT);

[0287] with the proviso that the second polynucleotide is not SEQ ID NO:6 (element X), and wherein the isolated polynucleotide is not SEQ ID NO:12 (PPE-1-3X).

[0288] According to some embodiments of the invention, each of the wild type M4, M5, M8, M6, M7 and/or M1 sequences is placed in a head to tail (5'→3') orientation with respect to the PPE-1 promoter set forth by SEQ ID NO:13.

[0289] According to some embodiments of the invention, each of the wild type M4, M5, M8, M6, M7 and/or M1 sequences is placed in a tail to head (3'→5') orientation with respect to the PPE-1 promoter set forth by SEQ ID NO:13.

[0290] According to some embodiments of the invention, the wild type M4, M5, M8, M6, M7 and/or M1 sequences are placed in various orientations (head to tail or tail to head) and/or sequential order with respect the other wild type M4, M5, M8, M6, M7 and/or M1 sequences, and/or with respect to the orientation of SEQ ID NO:13.

[0291] Construction of such viral vectors may be effected using known molecular biology techniques such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Springs Harbor Laboratory, New York (1989, 1992), in Ausubel et al., Current Protocols in Molecular Biology, John Wiley and Sons, Baltimore, Md. (1989), Chang et al., Somatic Gene Therapy, CRC Press, Ann Arbor, Mich. (1995), Vega et al., Gene Targeting, CRC Press, Ann Arbor Mich. (1995), Vectors: A Survey of Molecular Cloning Vectors and Their Uses, Butterworths, Boston Mass. (1988) and Gilboa et at. [Biotechniques 4 (6): 504-512, 1486].

[0292] Construction of the viral vector of SEQ ID NO: 9 is described in International Application WO/2008/132729, the contents of which are incorporated herein by reference.

[0293] As used herein, the term "administration" refers to providing or giving a subject an agent, such as an anti-angiogenic viral composition, by any effective route.

[0294] The viral vector of this aspect of the present invention may be administered per se or as part of a pharmaceutical composition which also includes a physiologically acceptable carrier. The purpose of a pharmaceutical composition is to facilitate administration of the active ingredient to an organism.

[0295] As used herein a "pharmaceutical composition" refers to a preparation of one or more of the active ingredients described herein with other chemical components such as physiologically suitable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration of a compound to an organism.

[0296] Herein the term "active ingredient" refers to the viral vector of the present invention accountable for the biological effect.

[0297] Hereinafter, the phrases "physiologically acceptable carrier" and "pharmaceutically acceptable carrier" which may be interchangeably used refer to a carrier or a diluent that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound. An adjuvant is included under these phrases.

[0298] Herein the term "excipient" refers to an inert substance added to a pharmaceutical composition to further facilitate administration of an active ingredient. Examples, without limitation, of excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils and polyethylene glycols.

[0299] Techniques for formulation and administration of drugs may be found in "Remington's Pharmaceutical Sciences," Mack Publishing Co., Easton, Pa., latest edition, which is incorporated herein by reference.

[0300] Suitable routes of administration may, for example, include oral, rectal, transmucosal, especially transnasal, intestinal or parenteral delivery, including intramuscular, intradermal, intraperitoneal, subcutaneous and intramedullary injections as well as intrathecal, direct intraventricular, intracardiac, e.g., into the right or left ventricular cavity, into the common coronary artery, intravenous, intraperitoneal, intranasal, or intraocular injections, sublingual, rectal, transdermal, intranasal, vaginal and inhalation routes. Injection of the viral vectors into a spinal fluid can also be used as a mode of administration.

[0301] The viral vectors or compositions thereof can be administered in an in-patient or out-patient setting. In one particular embodiment, the viral vectors or compositions thereof are administered in an injection or in an intravenous drip.

[0302] The present invention also contemplates engineering of the viral vectors in order to avoid, suppress or manipulate the immune response, ideally resulting in sustained expression and immune tolerance to the transgene product--such methods are described for example in Nayak et al., Gene Therapy (12 Nov. 2009), incorporated herein by reference.

[0303] Alternately, one may administer the pharmaceutical composition in a local rather than systemic manner, for example, via injection of the pharmaceutical composition directly into the tissue or tumor mass of a patient and even more directly into the tumor cells themselves.

[0304] Pharmaceutical compositions of the present invention may be manufactured by processes well known in the art, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or lyophilizing processes.

[0305] Pharmaceutical compositions for use in accordance with the present invention thus may be formulated in conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries, which facilitate processing of the active ingredients into preparations which, can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen.

[0306] For injection, the active ingredients of the pharmaceutical composition may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hank's solution, Ringer's solution, or physiological salt buffer. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.

[0307] For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

[0308] For administration by nasal inhalation, the active ingredients for use according to the present invention are conveniently delivered in the form of an aerosol spray presentation from a pressurized pack or a nebulizer with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichloro-tetrafluoroethane or carbon dioxide. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of, e.g., gelatin for use in a dispenser may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.

[0309] The pharmaceutical composition described herein may be formulated for parenteral administration, e.g., by bolus injection or continuous infusion. Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multidose containers with optionally, an added preservative. The compositions may be suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents.

[0310] Pharmaceutical compositions for parenteral administration include aqueous solutions of the active preparation in water-soluble form. Additionally, suspensions of the active ingredients may be prepared as appropriate oily or water based injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acids esters such as ethyl oleate, triglycerides or liposomes. Aqueous injection suspensions may contain substances, which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol or dextran. Optionally, the suspension may also contain suitable stabilizers or agents which increase the solubility of the active ingredients to allow for the preparation of highly concentrated solutions.

[0311] Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile, pyrogen-free water based solution, before use.

[0312] The pharmaceutical composition of the present invention may also be formulated in rectal compositions such as suppositories or retention enemas, using, e.g., conventional suppository bases such as cocoa butter or other glycerides.

[0313] Pharmaceutical compositions suitable for use in context of the present invention include compositions wherein the active ingredients are contained in an amount effective to achieve the intended purpose. More specifically, a therapeutically effective amount means an amount of active ingredients (i.e. viral particles) effective to prevent, alleviate or ameliorate symptoms of a disorder (e.g., thyroid cancer, neuroendocrine cancer) or prolong the survival of the subject being treated.

[0314] Determination of a therapeutically effective amount is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein. Therapeutic efficacy Of administration of the adenoviral vector of the present invention can be assessed according to a variety of criteria, including clinical presentation, biochemical parameters, radiological evaluation and the like. In some embodiments, efficacy is evaluated according to one or more of the following exemplary parameters:

[0315] Biodistribution:

[0316] for example, levels of virus DNA in blood and urine samples, expression of the fas-c transgene (mRNA) in blood;

[0317] Antibodies:

[0318] for example, levels of total anti-Ad-5 Ig, IgG and neutralizing anti-Ad5 antibodies in serum;

[0319] Angiogenic Biomarkers:

[0320] for example, von Willebrand Factor, TNFα, VEGFR1 and VEGFR2 levels in the blood;

[0321] Cytokine Levels:

[0322] for example, peripheral blood cytokine levels (e.g. IL-6, IL-8--see Table 6);

[0323] Tumor Response:

[0324] Tumor dimensions can be measured on CT (or MRI) scans, or other radiographic means. Tumor response can then be evaluated according to accepted criteria, such as Response Evaluation Criteria in Solid Tumors (RECIST).

[0325] The criteria can be evaluated at any time following administration, and can also be compared to pre-dosing values. In one embodiment, the evaluation criteria are assessed prior to administration of the adenovirus vector, and then on days 4±1, 7±1, 14±1, 28±2, day 56±3, day 112±4, about 3 months, about 4 months, about 5 months, about 6 months, about 1 year or more post dosing. In some embodiments, the evaluation criteria are assessed at day 7±1, 14±1, 28±2, day 56±3, day 112±4, about 3 months, about 6 months, about 9 months, about 1 year and about every 3 months thereafter for up to 2, up to 3, up to 4 or more years post dosing.

[0326] Determination of safety of dosing or dosing regimen is well within the ability of one skilled in the art. Safety can be assessed according to a variety of criteria, including, but not limited to, clinical presentation, tissue and organ pathology, presence of abnormal vital signs (e.g. pyrexia, fatigue, chills, tachycardia, hypertension, constipation and the like), hematology values (e.g. hemoglobin, hematocrit, RCV and the like), chemistry or urinalysis abnormalities (elevated enzymes such as alkaline phosphatase ALT, AST, bilirubin and the like) and ECG, EEG, etc.

[0327] As used herein, the term "antibody" refers to a molecule including an antigen binding site which specifically binds (immunoreacts with) an antigen. The term "antibody" includes intact molecules as well as functional fragments thereof, such as Fab, F(ab)2, and Fv that are capable of binding to macrophages. These functional antibody fragments are defined as follows: (1) Fab, the fragment which contains a monovalent antigen-binding fragment of an antibody molecule, can be produced by digestion of whole antibody with the enzyme papain to yield an intact light chain and a portion of one heavy chain; (2) Fab', the fragment of an antibody molecule that can be obtained by treating whole antibody with pepsin, followed by reduction, to yield an intact light chain and a portion of the heavy chain; two Fab' fragments are obtained per antibody molecule; (3) (Fab)2, the fragment of the antibody that can be obtained by treating whole antibody with the enzyme pepsin without subsequent reduction; F(ab)2 is a dimer of two Fab' fragments held together by two disulfide bonds; (4) Fv, defined as a genetically engineered fragment containing the variable region of the light chain and the variable region of the heavy chain expressed as two chains; and (5) Single chain antibody ("SCA"), a genetically engineered molecule containing the variable region of the light chain and the variable region of the heavy chain, linked by a suitable polypeptide linker as a genetically fused single chain molecule.

[0328] Methods of producing polyclonal and monoclonal antibodies as well as fragments thereof are well known in the art (See for example, Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, New York, 1988, incorporated herein by reference).

[0329] As used herein, the term "antigen" refers to a substance that can stimulate the production of antibodies or a T-cell response in a mammal, including compositions that are injected or absorbed into a mammal. An antigen reacts with the products of specific humoral or cellular immunity, including those induced by heterologous immunogens. The term "antigen" includes all related antigenic epitopes. In one example, an antigen is a cancer antigen. A target antigen is an antigen against which an immune response is desired, for example to achieve a therapeutic effect, such as tumor regression.

[0330] For any preparation used in the methods of the invention, the therapeutically effective amount or dose can be estimated initially from in vitro and cell culture assays. For example, a dose can be formulated in animal models to achieve a desired concentration or titer. Such information can be used to more accurately determine useful doses in humans.

[0331] The therapeutically effective amount of the active ingredient can be formulated in a unit dose. As used herein "unit dose" refers to a physically discrete unit containing a predetermined quantity of an active material calculated to individually or collectively produce a desired effect such as an anti-cancer effect. A single unit dose or a plurality of unit doses can be used to provide the desired effect, such as an anti-cancer therapeutic effect. In some embodiments, the unit dosage is 1×10⁸ to about 1×10¹⁶ virus particles, at least about 1×10¹¹ to about 1×10¹³ virus particles, and optionally about 1×10¹¹, about 3×10¹¹, about 5×10¹¹, about 1×10¹², about 3×10¹², about 5×10¹², about 1×10¹³, about 3×10¹³ or more virus particles.

[0332] Toxicity and therapeutic efficacy of the active ingredients described herein can be determined by standard pharmaceutical procedures in vitro, in cell cultures or experimental animals. The data obtained from these in vitro and cell culture assays and animal studies can be used in formulating a range of dosage for use in human. The dosage may vary depending upon the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dosage can be chosen by the individual physician in view of the patient's condition. (See e.g., Fingl, et al., 1975, in "The Pharmacological Basis of Therapeutics", Ch. 1 p. 1).

[0333] Dosage amount and interval may be adjusted individually to provide levels of the active ingredient sufficient to induce or suppress the biological effect (minimal effective concentration, MEC). The MEC will vary for each preparation, but can be estimated from in vitro data. Dosages necessary to achieve the MEC will depend on individual characteristics and route of administration. Detection assays can be used to determine plasma concentrations.

[0334] Depending on the severity and responsiveness of the condition to be treated, dosing can be of a single or a plurality of administrations, with course of treatment lasting from several days to several weeks or until cure is effected or diminution of the disease state is achieved.

[0335] The amount of a composition to be administered will, of course, be dependent on the subject being treated, the severity of the affliction, the manner of administration, the judgment of the prescribing physician, etc.

[0336] According to one embodiment, about 10³ to about 10¹⁶ virus particles are administered to the subject.

[0337] According to another embodiment, about 10⁵ to about 10¹³ virus particles are administered to the subject.

[0338] According to one embodiment, about 10⁷ to about 10¹² virus particles are administered to the subject.

[0339] According to one embodiment, about 1×10¹² to about 5×10¹² virus particles are administered to the subject.

[0340] According to yet another embodiment, about 1×10⁹ to about 1×10¹⁶ virus particles, at least about 1×10¹¹ to about 1×10¹³ virus particles are administered to the subject.

[0341] According to yet another embodiment the subject is administered intravenously with 1×10¹²-1×10¹³ viral particles of SEQ ID NO: 9, or SEQ ID NO: 10.

[0342] According to yet another embodiment the subject is administered intravenously with at least two doses of 1×10¹²-1×10¹³ viral particles of SEQ ID NO: 9, or SEQ ID NO: 10. According to yet another embodiment the subject is administered intravenously with at least three or more doses of 1×10¹²-1×10¹³ viral particles of SEQ ID NO: 9, or SEQ ID NO: 10. In a particular embodiment, the at least two doses are administered at least about 1 day, at least about 3 days, at least about 5 days, at least about 7 days, at least about 2 weeks, at least about 3 weeks, at least about 4 weeks, at least about 2 months, at least about 6 months, at least about 9 months, at least about 1 year, at least about 1.25 years, at least about 1.5 years, at least about 1.75 years, at least about 2 years, at least about 2.5 years, at least about 3 years or more apart.

[0343] Compositions of the present invention may, if desired, be presented in a pack or dispenser device, such as an FDA approved kit, which may contain one or more unit dosage forms containing the active ingredient. The pack may, for example, comprise metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also be accommodated by a notice associated with the container in a form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the compositions or human or veterinary administration. Such notice, for example, may be of labeling approved by the U.S. Food and Drug Administration for prescription drugs or of an approved product insert. Compositions comprising a preparation of the invention formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition, as is further detailed above.

[0344] Thus, according to some aspects of some embodiments of the present invention there is provided a kit for treating a solid tumor in a subject in need thereof, comprising a unit dosage of virus particles of a non-replicating adenovirus vector comprising a polynucleotide having a nucleotide sequence as set forth in SEQ ID NOs: 9 or 10, wherein the non-replicating adenovirus vector is formulated for intravenous administration, and instructions for administration of the adenovirus.

[0345] The adenovirus vector can be used in combination with other treatments. For example, the uptake of adenoviral vectors into EC cells can be enhanced by treating the vectors with engineered antibodies or small peptides. Such "adenobody" treatment, was shown effective in directing adenovirus constructs to EGF receptors on cells (Watkins et al 1997, Gene Therapy 4:1004-1012). In addition, Nicklin et al have shown that a small peptide, isolated via phage display, increased specificity and efficiency of vectors in endothelial cells and decreased the expression in liver cells in culture (Nicklin et al 2000, Circulation 102:231-237). In a recent study, an FGF retargeted adenoviral vector reduced the toxicity of tk in mice (Printz et al 2000, Human Gene Therapy 11:191-204).

[0346] Low dose radiation has been shown to cause breaks in DNA strands primarily in the G2/M phase, cell membrane damage enhancing the bystander effect, and thus may potentiate other cytotoxic and anti-neoplastic therapies, when administered in combination. Vascular endothelial cells may be particularly suitable to such combination, or adjunct, therapies, since it has been demonstrated that low dose radiation specifically targets the apoptotic system of the microvascular endothelial cells (Kolesnick et al., Oncogene 2003; 22:5897-906). Angiostatin has been shown to potentiate the therapeutic effects of low dose radiation (Gorski et al. Can Res 1998; 58:5686-89). However, the effects of radiation are still poorly understood, since irradiation has also been shown to increase pro-angiogenic "tissue repair factors" (Itasaka et al., Am Assoc Canc Res, 2003; abstract 115). Similarly, certain chemotherapeutic agents have been shown to activate specific cytotoxic and apoptotic pathways [doxorubicin, cisplatin and mitomycin C induce accumulation of Fas receptor, FADD, and other proapoptotic signals in the FADD/MORT-1 pathway (Micheau et al., BBRC 1999 256:603-07)].

[0347] For example International Application WO/2008/132729 teaches combined doxorubicin and AdPPE-1 (3x)-Fas-c chimera construct administration in endothelial cells (BAEC). Thus, the viral vectors and the pharmaceutical compositions comprising same of the present invention can be used to treat solid tumors alone or in combination with one or more other established or experimental therapeutic regimen for such disorders. Therapeutic regimen for treatment of solid tumors suitable for combination with the viral vectors of the present invention include, but are not limited to chemotherapy, radiotherapy, phototherapy and photodynamic therapy, surgery, nutritional therapy, ablative therapy, combined radiotherapy and chemotherapy, brachiotherapy, proton beam therapy, immunotherapy, cellular therapy and photon beam radiosurgical therapy. The vectors of the present invention may be administered with additional ingredients which may improve the uptake of the nucleic acid construct by the cells, expression of the chimeric polypeptide by the nucleic acid construct in the cells, the activity of the expressed chimeric polypeptide or the efficacy of the treatment on any aspects of the disease. Protocols for use of recombinant anti-angiogenic adenovirus vectors for cancer treatment are known in the art. Many clinical trials of adenovirus-based anti-angiogenic gene therapy are currently being conducted, mostly involving a recombinant anti-angiogenic adenovirus in combination with other cancer therapies, and administered intra-tumorally, such as an adenovirus-p53 vaccine with chemotherapy for small cell lung cancer (NCT0049218), adenovirus-suicide gene with chemotherapy for small cell lung cancer (NCT00964756), an adenovirus-endostatin construct with chemotherapy for head and neck cancer (NCT00634595), an adenovirus-suicide gene with chemotherapy for malignant melanoma (NCT00005057) and an adenovirus-tk construct with chemotherapy for hepatocellular carcinoma (NCT00844623).

[0348] Anti-cancer drugs that can be co-administered with the compounds of the invention include, but are not limited to Acivicin; Aclarubicin; Acodazole Hydrochloride; Acronine; Adriamycin; Adozelesin; Aldesleukin; Altretamine; Ambomycin; Ametantrone Acetate; Aminoglutethimide; Amsacrine; Anastrozole; Anthramycin; Asparaginase; Asperlin; Azacitidine; Azetepa; Azotomycin; Batimastat; Benzodepa; Bicalutamide; Bisantrene Hydrochloride; Bisnafide Dimesylate; Bevacizumab, Bizelesin; Bleomycin Sulfate; Brequinar Sodium; Bropirimine; Busulfan; Cactinomycin; Calusterone; Caracemide; Carbetimer; Carboplatin; Carmustine; Carubicin Hydrochloride; Carzelesin; Cedefingol; Chlorambucil; Cirolemycin; Cisplatin; Cladribine; Crisnatol Mesylate; Cyclophosphamide; Cytarabine; Dacarbazine; Dactinomycin; Daunorubicin Hydrochloride; Decitabine; Dexormaplatin; Dezaguanine; Dezaguanine Mesylate; Diaziquone; Docetaxel; Doxorubicin; Doxorubicin Hydrochloride; Droloxifene; Droloxifene Citrate; Dromostanolone Propionate; Duazomycin; Edatrexate; Eflornithine Hydrochloride; Elsamitrucin; Enloplatin; Enpromate; Epipropidine; Epirubicin Hydrochloride; Erbulozole; Esorubicin Hydrochloride; Estramustine; Estramustine Phosphate Sodium; Etanidazole; Etoposide; Etoposide Phosphate; Etoprine; Fadrozole Hydrochloride; Fazarabine; Fenretinide; Floxuridine; Fludarabine Phosphate; Fluorouracil; Fluorocitabine; Fosquidone; Fostriecin Sodium; Gemcitabine; Gemcitabine Hydrochloride; Hydroxyurea; Idarubicin Hydrochloride; Ifosfamide; Ilmofosine; Interferon Alfa-2a; Interferon Alfa-2b; Interferon Alfa-n1; Interferon Alfa-n3; Interferon Beta-I a; Interferon Gamma-I b; Iproplatin; Irinotecan Hydrochloride; Lanreotide Acetate; Letrozole; Leuprolide Acetate; Liarozole Hydrochloride; Lometrexol Sodium; Lomustine; Losoxantrone Hydrochloride; Masoprocol; Maytansine; Mechlorethamine Hydrochloride; Megestrol Acetate; Melengestrol Acetate; Melphalan; Menogaril; Mercaptopurine; Methotrexate; Methotrexate Sodium; Metoprine; Meturedepa; Mitindomide; Mitocarcin; Mitocromin; Mitogillin; Mitomalcin; Mitomycin; Mitosper; Mitotane; Mitoxantrone Hydrochloride; Mycophenolic Acid; Nocodazole; Nogalamycin; Ormaplatin; Oxisuran; pazotinib; Paclitaxel; Pegaspargase; Peliomycin; Pentamustine; Peplomycin Sulfate; Perfosfamide; Pipobroman; Piposulfan; Piroxantrone Hydrochloride; Plicamycin; Plomestane; Porfimer Sodium; Porfiromycin; Prednimustine; Procarbazine Hydrochloride; Puromycin; Puromycin Hydrochloride; Pyrazofurin; Riboprine; Rogletimide; Safingol; Safingol Hydrochloride; Semustine; Simtrazene; Sorafinib; Sparfosate Sodium; Sparsomycin; Spirogermanium Hydrochloride; Spiromustine; Spiroplatin; Streptonigrin; Streptozocin; Sulofenur; Sunitinib; Talisomycin; Taxol; Tecogalan Sodium; Tegafur; Teloxantrone Hydrochloride; Temoporfin; Teniposide; Teroxirone; Testolactone; Thiamiprine; Thioguanine; Thiotepa; Tiazofuirin; Tirapazamine; Topotecan Hydrochloride; Toremifene Citrate; Trestolone Acetate; Triciribine Phosphate; Trimetrexate; Trimetrexate Glucuronate; Triptorelin; Tubulozole Hydrochloride; Uracil Mustard; Uredepa; Vapreotide; Verteporfin; Vinblastine Sulfate; Vincristine Sulfate; Vindesine; Vindesine Sulfate; Vinepidine Sulfate; Vinglycinate Sulfate; Vinleurosine Sulfate; Vinorelbine Tartrate; Vinrosidine Sulfate; Vinzolidine Sulfate; Vorozole; Zeniplatin; Zinostatin; Zorubicin Hydrochloride. Additional antineoplastic agents include those disclosed in Chapter 52, Antineoplastic Agents (Paul Calabresi and Bruce A. Chabner), and the introduction thereto, 1202-1263, of Goodman and Gilman's "The Pharmacological Basis of Therapeutics", Eighth Edition, 1990, McGraw-Hill, Inc. (Health Professions Division).

[0349] The present inventors have shown that administering a single dose of the viral vectors of the present invention (e.g. Ad5PPE-1-3X fas-chimera) in combination with the chemotherapeutic drug sunitinib (Sutent), can improve the efficacy of the chemotherapy treatment, or of the viral vector's effect on a metastatic cancer, in the Lewis Lung Carcinoma model (see Example VI below). Thus, in some embodiments the viral vectors of the present invention are administered in combination with one or more of the chemotherapeutic drugs, such as sunitinib (Sutent).

[0350] Chemotherapeutic agents can be administered along with the viral vectors of the invention, prior to treatment with the viral vectors of the present invention, or following treatment with the viral vectors of the present invention. In a particular embodiment, the chemotherapeutic agent is administered at least about 1 day, at least about 3 days, at least about 5 days, at least about 7 days, at least about 2 weeks, at least about 3 weeks, at least about 4 weeks, at least about 2 months, at least about 6 months, at least about 9 months, at least about 1 year prior to initiation of treatment with the viral vector of the present invention. In another particular embodiment, the chemotherapeutic agent is administered at least about 1 day, at least about 3 days, at least about 5 days, at least about 7 days, at least about 2 weeks, at least about 3 weeks, at least about 4 weeks, at least about 2 months, at least about 6 months, at least about 9 months, at least about 1 year following initiation of treatment with the viral vector of the present invention. In another particular embodiment, the chemotherapeutic agent is administered during, or alongside with initiation of treatment with the viral vector of the present invention.

[0351] The viral vectors of the present invention may also be administered with an agent that enhances expression of transgenes in adenoviral-mediated transient expression. For example International Application WO/2008/132729 teaches administration of a corticosteroid (e.g. dexamethasone and/or N-Acetyl Cysteine (NAC) prior to AdPPE-1 (3x)-Fas-c chimera construct administration, or concomitant treatment with an endothelin inhibitor such as bosentan.

[0352] In addition, the viral vectors of the present invention may also be administered with an agent that brings about transient immunosuppression, such as for example deoxyspergualin (DSG) or cyclophosphamide (see for example Smith et al., Gene Ther. 1996 June; 3(6):496-502) in order to allow for repetitive dosing.

[0353] Adenovirus-based gene therapy protocols have commonly been limited to single doses, for reasons pertaining to safety and efficacy of the drug, particularly concerning patient anti-adenovirus immune response, as most populations are repeatedly exposed and sensitized to various adenoviral antigens. The present inventors have surprisingly found no correlation between levels of anti-adenovirus antibodies in serum of patients following administration of the adenovirus of the invention, and the dose of adenovirus administered. Evaluation of antibody titers, including anti-Ad5 neutralizing antibodies, IgG and total anti-Ad5 antibodies in the patients (see Example II, and FIGS. 2 and 3A-3B) revealed no effect of the baseline levels of neutralizing anti-adenovirus antibody on disease progression following dosing. Evaluation of total anti-adenovirus Ad5 antibody titers, and specific anti-Ad5 IgG in serum of patients prior to, and following administration of the Ad5-PPE-1-3X-fas-c adenoviral vectors revealed increased antibody titers following adenovirus vector administration, but indicated no correlation with the doses administered (see Table 4 in Example II that follows), suggesting that repeated doses of the adenovirus as described herein may not lead to an immune response limiting the clinical utility of the adenovirus in the host. Further, it was found that administration of AdSPPE-1-3X-fas chimera was effective in reducing disease progression even in subjects with high levels of neutralizing anti-Ad5 antibodies and total anti-Ad5 antibodies detected prior to first injection of the construct (see FIGS. 2 and 3). Yet further, assays of blood and urine Ad5-PPE-1-3X-fas-c levels (biodistribution) following administration revealed high levels of the adenovirus constructs of the invention 28 days post-administration, even in the presence of elevated total or IgG anti-Ad5 antibody levels (see Tables 4 and 7 of Example II that follows, specifically, see subjects 2, 9 and 10). In one subject [036], for example, significant levels of transgene expression were detected in an aspirate from a metastatic lesion as many as 28 days following adenovirus vector administration, despite a massive fold increase (X3125) in anti-Ad5 antibody titer (see Tables 4 and 9 in Example II that follows).

[0354] Thus, in some embodiments, administering the dose of the adenovirus vector of the present invention inhibits growth of a tumor. In other embodiments, administering the adenovirus vector inhibits angiogenesis of the tumor.

[0355] Thus, the adenovirus vectors of the present invention can be administered in one, at least two, three or more doses, with intervals therebetween sufficient for antibody formation, without causing a dose-dependent antiviral antibody response. Such intervals are typically 21-28 days, but may be as few as 1 or 2 days, or as many as 7 days, 10 days, 2 weeks, three weeks, four, six, eight ten or more weeks. Thus, according to another aspect of some embodiments of the present invention there is provided a method for administering a therapeutically effective amount of a therapeutic composition comprising an adenoviral vector to a subject in need thereof comprising administering the composition to the subject at least twice, wherein the administration does not induce a dose-dependent increase in antibodies against the adenoviral vector in the subject. According to another aspect of one embodiment of the present invention, the time between administration between a first dose and an at least second dose is sufficient for anti-Ad5 antibody formation.

[0356] In yet another embodiment, the adenovirus vectors of the present invention are effective when administered to subjects having elevated levels of serum anti Ad5 antibodies. In some particular embodiments the anti-Ad5 antibody levels are elevated compared to the subjects pre-dosing baseline anti-Ad5 levels. In yet other embodiments, the adenovirus vectors of the present invention are detected in the blood of the subjects having elevated anti-Ad5 antibody levels are compared to the subjects pre-dosing baseline anti-Ad5 levels, at day 4 post-administration, day 7 post-administration, day 15 post-administration, day 21 post-administration, day 28 post-administration, day 37 post-administration, day 56 post-administration or day 112 post-administration or more. Such anti-Ad5 antibodies can be neutralizing antibodies, total anti-Ad5 antibodies, or a specific anti-Ad5 antibody subtype, such as anti-Ad5 IgG.

[0357] Thus, in some embodiments, the adenovirus is detected in the blood of the subject at least about 4 days post administration. According to other embodiments an amount of serum anti-adenovirus antibodies is increased following the administering, and the adenovirus is detected in the blood of the subject at least about 21 days post administration.

[0358] It is expected that during the fife of a patent maturing from this application many relevant chemotherapeutic agents will be developed and the scope of the term chemotherapeutic agent is intended to include all such new technologies a priori.

[0359] As used herein the term "about" refers to ±10%.

[0360] The terms "comprises", "comprising", "includes", "including", "having" and their conjugates mean "including but not limited to".

[0361] The term "consisting of means "including and limited to".

[0362] The term "consisting essentially of" means that the composition, method or structure may include additional ingredients, steps and/or parts, but only if the additional ingredients; steps and/or parts do not materially alter the basic and novel characteristics of the claimed composition, method or structure.

[0363] As used herein, the singular form "a", "an" and "the" include plural references unless the context clearly dictates otherwise. For example, the term "a compound" or "at least one compound" may include a plurality of compounds, including mixtures thereof.

[0364] Throughout this application, various embodiments of this invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

[0365] Whenever a numerical range is indicated herein, it is meant to include any cited numeral (fractional or integral) within the indicated range. The phrases "ranging/ranges between" a first indicate number and a second indicate number and "ranging/ranges from" a first indicate number "to" a second indicate number are used herein interchangeably and are meant to include the first and second indicated numbers and all the fractional and integral numerals therebetween.

[0366] As used herein the term "method" refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by practitioners of the chemical, pharmacological, biological, biochemical and medical arts.

[0367] As used herein, the term "treating" includes abrogating, substantially inhibiting, slowing or reversing the progression of a condition, substantially ameliorating clinical or aesthetical symptoms of a condition or substantially preventing the appearance of clinical or aesthetical symptoms of a condition.

[0368] It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination or as suitable in any other described embodiment of the invention. Certain features described in the context of various embodiments are not to be considered essential features of those embodiments, unless the embodiment is inoperative without those elements.

[0369] Various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below find experimental support in the following examples.

EXAMPLES

[0370] Reference is now made to the following examples, which together with the above descriptions illustrate some embodiments of the invention in a non limiting fashion.

[0371] Generally, the nomenclature used herein and the laboratory procedures utilized in the present invention include molecular, biochemical, microbiological and recombinant DNA techniques. Such techniques are thoroughly explained in the literature. See, for example, "Molecular Cloning: A laboratory Manual" Sambrook et al., (1989); "Current Protocols in Molecular Biology" Volumes I-III Ausubel, R. M., ed. (1994); Ausubel et al., "Current Protocols in Molecular Biology", John Wiley and Sons, Baltimore, Md. (1989); Perbal, "A Practical Guide to Molecular Cloning", John Wiley & Sons, New York (1988); Watson et al., "Recombinant DNA", Scientific American Books, New York; Birren et al. (eds) "Genome Analysis: A Laboratory Manual Series", Vols. 1-4, Cold Spring Harbor Laboratory Press, New York (1998); methodologies as set forth in U.S. Pat. Nos. 4,666,828; 4,683,202; 4,801,531; 5,192,659 and 5,272,057; "Cell Biology: A Laboratory Handbook", Volumes I-III Cellis, J. E., ed. (1994); "Culture of Animal Cells--A Manual of Basic Technique" by Freshney, Wiley-Liss, N.Y. (1994), Third Edition; "Current Protocols in Immunology" Volumes I-III Coligan J. E., ed. (1994); Stites et al. (eds), "Basic and Clinical Immunology" (8th Edition), Appleton & Lange, Norwalk, Conn. (1994); Mishell and Shiigi (eds), "Selected Methods in Cellular Immunology", W. H. Freeman and Co., New York (1980); available immunoassays are extensively described in the patent and scientific literature, see, for example, U.S. Pat. Nos. 3,791,932; 3,839,153; 3,850,752; 3,850,578; 3,853,987; 3,867,517; 3,879,262; 3,901,654; 3,935,074; 3,984,533; 3,996,345; 4,034,074; 4,098,876; 4,879,219; 5,011,771 and 5,281,521; "Oligonucleotide Synthesis" Gait, M. J., ed. (1984); "Nucleic Acid Hybridization" Hames, B. D., and Higgins S. J., eds. (1985); "Transcription and Translation" Hames, B. D., and Higgins S. J., eds. (1984); "Animal Cell Culture" Freshney, R. I., ed. (1986); "Immobilized Cells and Enzymes" IRL Press, (1986); "A Practical Guide to Molecular Cloning" Perbal, B., (1984) and "Methods in Enzymology" Vol. 1-317, Academic Press; "PCR Protocols: A Guide To Methods And Applications", Academic Press, San Diego, Calif. (1990); Marshak et al., "Strategies for Protein Purification and Characterization--A Laboratory Course Manual" CSHL Press (1996); all of which are incorporated by reference as if fully set forth herein. Other general references are provided throughout this document. The procedures therein are believed to be well known in the art and are provided for the convenience of the reader. All the information contained therein is incorporated herein by reference.

Example I

Efficacy of AdSPPE-1-3X-fas chimera in Lewis Lung Carcinoma Model

[0372] In order to evaluate effective dosage ranges Ad5PPE-1-3X-fas chimera adenovirus for treatment of solid tumors, metastatic tumors were induced in the mouse Lewis Lung Carcinoma model, and a range of doses of Ad5PPE-1-3X-fas chimera adenovirus (SEQ ID NO: 10) administered systemically.

Materials and Experimental Methods

[0373] Construction and Cloning of the Viral Vector:

[0374] The vector was constructed using a backbone containing most of the genome of adenovirus type 5, as well as partial homology to an adaptor plasmid, which enables recombination.

[0375] The E1 early transcriptional unit was deleted from the backbone plasmid, and further modified by deleting the pWE25 and the Amp resistance selection marker site.

[0376] The adaptor plasmid, containing sequences of the Ad5, CMV promoter, MCS, and SV40 polyA was modified to delete deleting the CMV promoter, and the PPE-1 promoter and Fas-c fragment were inserted by restriction digestion.

[0377] The modified PPE-1 promoter (PPE-1-3X, SEQ ID NO: 12) and the Fas-chimera transgene (Fas-c, SEQ ID NO: 4) were utilized for construction of the adenoviral vector. The PPE-1-(3X)-Fas-c element (2115 bp) was constructed from the PPE-1-(3X)-luc element. This element contains the 1.4 kb of the murine preproendothelin PPE-1-(3X) promoter, the Luciferase gene, the SV40 polyA site and the first intron of the murine ET-1 gene, originated from the pEL8 plasmid (8848 bp) used by Harats et al (Harats D. et al., JCI, 1995). The PPE-3-Luc cassette was extracted from the pEL8 plasmid using the BamHI restriction enzyme. The Luciferase gene was substituted by the Fas-c gene [composed of the extra cellular and intra membranal domains of the human TNF-R1 (Tumor Necrosis Factor Receptor 1, SEQ ID NO: 2) and of the Fas (p55) intracellular domain (SEQ ID NO: 3) (Boldin et al, JBC, 1995)] to obtain the PPE-1-3x-Fas-c cassette.

[0378] PPE-1(3x)-Fas-c Plasmid--The cassette was further introduced into the backbone plasmid by restriction digestion, resulting with the PPE-1(3x)-Fas-c plasmid.

[0379] Adaptor-PPE-1(3x)-Fas-c Plasmid--The PPE-1-3x-Fas-c element was extracted from the first generation construct PPE-1-3x-Fas-c plasmid, and was amplified with designated PCR primers introducing SnaB1 and EcoR1 restriction sites at the 5'-and-3'-end respectively. These sites were used to clone the PPE-Fas-c fragment into the adaptor plasmid digested with SnaB1 and EcoR1, resulting in the adaptor-PPE-1-3x-Fas-c used for transfection of the host cells (for example, PER.C6 cells).

[0380] Lewis Lung Carcinoma Model

[0381] Freshly harvested D122 Lewis Lung Carcinoma (LLC) cells (5×10⁵ per mouse) were administered to C57BL/6 mice (3 months old), by subcutaneous injection into the foot pad. Primary tumors developed in the feet in about 14 days. When tumors reached a diameter of at least 7 mm, the mice were anesthetized and the distal segment of the limb was amputated. Five days after limb amputation, mice were randomized to one of the following groups:

TABLE-US-00001 Dose- virus particles Treatment (vp/mouse) Group 1 Ad5PPE-1-3X-fas chimera adenovirus .sup. 10¹¹ Group 2 Ad5PPE-1-3X-fas chimera adenovirus .sup. 10¹⁰ Group 3 Ad5PPE-1-3X-fas chimera adenovirus 10⁹ Group 4 Ad5PPE-1-3X-fas chimera adenovirus 10⁸ Group 5 Ad5PPE-1-3X-fas chimera adenovirus 10⁷ Group 6 Ad5PPE-1-3X-fas chimera adenovirus 10⁶ Group 7 Vehicle 0

[0382] Vehicle=PBS, 10% glycerol

[0383] Viruses were intravenously injected to the mouse tail. Mice were sacrificed upon death of 5 control (vehicle injected) mice (approximately 24 days after virus/control administration). Mice in which the primary tumor re-emerged after treatment, were withdrawn from the study. Upon sacrifice, animal's lungs were removed, washed and weighed, and blood was collected for liver function testing.

[0384] Liver and tumor tissues were cut and frozen in 4% formaldehyde or in OCT compound for histological analysis by hematoxylin-eosin staining or anti PECAM1 (anti-CD31) staining, respectively. Tumor weight differences were evaluated using the Mann-Whitney U-test.

[0385] The purpose of this study was to evaluate the dose dependent efficacy of Ad5PPE-1-3X-fas chimera adenovirus and to assess the minimal effective dose.

Results

[0386] Clinically Significant Doses (10⁶ to 10¹¹ vp/Mouse) of Ad5PPE-1-3X-fas Chimera Adenovirus do not Cause Non-Specific Systemic Effects

[0387] Mortality was observed in all groups prior to scheduled sacrifice with no dose response trend. Clinical signs of surviving mice did not reveal any abnormalities. No dose response trend was observed in mice weights.

[0388] No statistical significant differences were observed in the kidney function in all groups, except for an increase in uric acid observed at the lowest dose cohort on day 5 post dosing. No statistical significant differences were observed in the SGOP and SGPT levels of treated vs. non treated mice.

[0389] Macroscopic examination at necropsy (brain, liver heart, spleen, gonads, kidneys, small intestine, lungs), did not reveal any abnormalities except for metastases found in the lungs. No statistical differences were found in the weight of organs between groups (liver, spleen, small intestine, heart, kidney, lung, and brain), except for the heart and spleen where an increase (not dose dependent) in the weight was observed in mice treated with adenovirus compared to the untreated mice.

[0390] Microscopic observations (on the lung, liver and heart) showed mild to moderate inflammation and regenerative changes of the liver following treatment with 10⁸-10¹¹ vp/mouse.

[0391] Ad5PPE-1-3X-fas Chimera Adenovirus (10⁸ to 10¹¹ vp/Mouse) Suppresses Metastatic Tumor Growth

[0392] FIGS. 1A-1C show exemplary lungs of treatment and control groups, at the completion of the study. In the lung, large metastases were found in mice treated with vehicle (FIG. 1A). These metastases were reduced in a dose dependent fashion in Ad5PPE-1-3X-fas chimera adenovirus-treated mice (FIGS. 1B-1C), with the strongest effect in the highest dose (10¹¹ vp/mouse) cohort (FIG. 1C), in which most of the lungs appeared to be normal or with only small metastases. This effect decreased to non observable levels at low titers of the virus (10⁷ vp/mouse and 10⁶ vp/mouse). The protective, anti-tumor effect of the AdSPPE-1-3X-fas chimera adenovirus was confirmed by the difference in weight of the lungs of the Ad5PPE-1-3X-fas chimera adenovirus-treated mice, which was significantly reduced when compared to control mice but diminished to non observable level at low virus titers (FIG. 1D).

[0393] Thus, a single, systemic injection of a clinically significant amount of Ad5PPE-1-3X-fas chimera adenovirus was highly effective in reducing the tumor burden in Lewis Lung Carcinoma model in a dose dependent fashion. 10¹¹ vp/mouse resulted in a 70% decrease in tumor burden. No efficacy of treatment was observed at the lowest doses (10⁶ vp/mouse-10⁷ vp/mouse).

Example II

Administration of Ad5PPE-1-3X-fas chimera in the Clinical Setting

[0394] In order to determine the safety and efficacy of administration of AD5PPE-1-3X-fas-chimera in the clinical setting, outcomes such as toxicity, adverse effects, antibody titer, biodistribution, disease progression and disease recurrence and survival were monitored in subjects with solid primary and metastatic tumors receiving intravenous infusion of a range of doses of the Ad5PPE-1-3X-fas chimera adenovirus vector.

Materials and Experimental Methods

[0395] Subject Population:

[0396] Subjects with advanced and/or metastatic solid organ cancer 18 years of age and older, without remaining options for standard curative or palliative measures are enrolled.

[0397] Criteria for subject's inclusion in the treatment group are:

[0398] 1. Subjects at least 18 years of age;

[0399] 2. Histologically confirmed malignancy that is metastatic or unresectable and for which standard curative or palliative measures do not exist or are no longer effective;

[0400] 3. Karnofsky performance status of ≧70%;

[0401] 4. Adequate haematological profile: ANC≧1500/μl, hemoglobin≧10 g/dl, platelets>100,000/μl, and INR within normal limits;

[0402] 5. Adequate renal function (CCT>60 mL/min/1.73 m2);

[0403] 6. Adequate hepatic function: (ALT and AST<2.5×ULN) and total bilirubin within normal limits.

[0404] Criteria for subject's exclusion from the treatment group are:

[0405] 1. Recent cardiac event (within 12 months) or active cardiac/vascular disease;

[0406] 2. Recent surgery (within 4 weeks);

[0407] 3. Proliferative retinopathy;

[0408] 4. Liver disease;

[0409] 5. CNS metastasis;

[0410] 6. Recent anti-angiogenic therapy (within 6 weeks);

[0411] 7. Current/recent (within 4 weeks) immunosuppressive therapy;

[0412] 8. Recent chemo/radiotherapy or investigational agent (within 4 weeks);

[0413] 9. Uncontrolled co-morbidity.

[0414] Composition: Ad5-PPE-1-3X-fas-chimera

[0415] Ad5-PPE-1-3X-fas-chimera (SEQ ID NO: 10) is a vascular disruptive gene therapeutic, consisting of a non-replicating adenovirus vector (Ad5, E1 deleted, SEQ ID NO: 1) which contains a modified murine pre-proendothelin promoter (PPE-1-3x, SEQ ID NO: 12) and a fas-chimera transgene [Fas and human tumor necrosis factor (TNF) receptor](SEQ ID NO: 4). It is formulated as a sterile vector solution and supplied frozen (below -65° C.), in single use vials. Each vial contains 0.5 mL of vector solution at a specific viral titer.

[0416] Dosage

[0417] Dose escalation is done by cohort as follows:

TABLE-US-00002 Dose (virus particles) Subjects per Cohort Cohort 1 1 × 10¹⁰ 3 Cohort 2 3 × 10¹⁰ 3 Cohort 3 1 × 10¹¹ 3 Cohort 4 3 × 10¹¹ 3 Cohort 5 1 × 10¹² 3 Cohort 6 3 × 10¹² 12 Cohort 7 1 × 10¹³ 6

[0418] Since no dose limiting toxicity was observed after dosing 3 subjects in each of the first 6 cohorts, 9 additional subjects were enrolled into cohort 6 according to the protocol and after approval of the Institutional Review Boards. 27 subjects (3 in each of cohorts 1-5 [total of 15 subjects] and 12 in Cohort 6) were treated. Since no dose limiting toxicity was observed, 6 more patients were enrolled to cohort 7. A total of 33 subjects were treated. One case of NCI Grade 3 fever, shortly following administration of the highest dose (cohort 7), was observed.

[0419] The Ad5-PPE-1-3X-fas-chimera is administered as an intravenous infusion. The same drug volume and saline volume were used for each subject within each cohort, and each subject was infused with the same volume of the drug. The infusion duration was between 3 and 5 minutes in cohorts 1-5 and 15 minutes in cohort 6 and 50 minutes for cohort 7. In general, these numbers reflect the instructions specified in the pharmacy manual provided with the study.

[0420] Subject Evaluation Criteria

[0421] Biodistribution:

[0422] Blood and urine samples were collected prior to dosing, at the end of the infusion (blood samples only), 3 hours, 6 hours, and on days 4(±1), 7(±1), 14(±1), 28(±2) and at day 56(±3) for evaluation of levels of virus DNA (in blood and urine) and the expression of the transgene (messenger RNA in blood). Only samples with detectable viral DNA are tested for transgene expression.

[0423] Antibodies:

[0424] Serum samples were collected prior to dosing for analysis of total anti-Ad-5 Ig, IgG and neutralizing anti-Ad5 antibody levels, and on day 28 and day 56 for analysis of total anti-Ad-5 Ig, IgG anti-Ad5 antibody levels.

[0425] Ad-5-IgG assay: Serum samples were diluted and analyzed for adenovirus-specific immunoglobulin G (IgG) by ELISA. For the ELISA, 96 well flat bottomed, high-binding Immulon-IV plates were coated with 50 ul Ad5-antigen (Upenn Vector Core) at (5×10 8 particles/well) in pH 9.5 carbonate buffer (coating buffer) overnight at 4° C., washed two times in PBS/0.05% Tween, blocked in PBS/1% HSA for 1 hour at 24° C., and then washed two times in PBS/0.05 Tween. Appropriately diluted (3-fold) samples were added to antigen-coated plates and incubated for 4 hours at room temperature. Plates were washed three times with PBS/0.05% Tween and incubated with peroxidase conjugated goat anti-human Ig (1:5000 dilution, Jackson ImmunoResearch Laboratories, Inc.) for 1 hour at room temperature. Plates were washed three times as above and TMB substrate (Sigma Chemical Co., St Louis, Mo.) is added at 100 ul/well for 30 minutes. Reaction was stopped when 100 ul 2N H₂SO₄ was added, and optical densities were read at 450 nm on a VersaMAX tunable microplate reader (Molecular Devices). Titer is the dilution achieving 0.5 maximum OD.

[0426] Neutralizing anti-Ad5 antibodies assay: The ability of serum to block adenovirus infection of Hela cells (ATCC) in vitro was analyzed utilizing adenovirus expressing green fluorescent protein (GFP) as a reporter (Upenn Vector Core). Various dilutions of test sera and, as a control/standard, pooled AB sera (Sigma), pre-incubated with moi of 1000 of reporter viruses for 1 hour at 37° C. were added to 90% confluent cell cultures. Cells were incubated for 16 hours and expression of GFP quantified by fluoroimaging using the Victor² (Wallace/PerkinElmer) and visually. The neutralizing titer of the sera was calculated as the reciprocal dilution of serum with 50% of the maximum fluorescence at 1000 MOI.

[0427] Angiogenic Biomarkers:

[0428] Blood samples were collected prior to dosing, and at the following visits thereafter, for evaluation of von Willebrand Factor levels and TNFα levels, on days 4±1, 7±1, 14±1, 28±2 and at day 56±3 post dosing.

[0429] Cytokine Levels:

[0430] Peripheral blood cytokine (see Table 6 below) levels were measured in cohort 6 patients at baseline, and at the following times after dosing with Ad5-PPE-1-3X-fas-chimera, at 6 hours, 4 days, 7 days, 14 days, 28 days, and 56 days post dosing.

[0431] Tumor Response:

[0432] The possible effect of the drug treatment on tumor response was evaluated by measuring the tumor dimensions on CT (or MRI) scans according to Response Evaluation Criteria in Solid Tumors (RECIST) criteria at screening and at week 4 and 8 post dosing, or according to RECIST criteria at screening, baseline day -1/0 and at day 7±1, week 4 (day 28±2) and week 8 (day 56±3) post dosing.

[0433] RECIST Criteria:

Evaluation of Target Lesions

[0434] Complete Response (CR):

[0435] Disappearance of all target lesions.

[0436] Partial Response (PR):

[0437] At least a 30% decrease in the sum of the LD of target lesions, taking as reference the baseline sum LD.

[0438] Stable Disease (SD):

[0439] Neither sufficient shrinkage to qualify for PR nor sufficient increase to qualify for PD, taking as reference the smallest sum LD since the treatment started.

[0440] Progressive Disease (PD):

[0441] At least a 20% increase in the sum of the LD of target lesions, taking as reference the smallest sum LD recorded since the treatment started or the appearance of one or more new lesions

Evaluation of Non-Target Lesions

[0442] Complete Response (CR):

[0443] Disappearance of all non-target lesions and normalization of tumor marker level

[0444] Incomplete Response/Stable Disease (SD):

[0445] Persistence of one or more non-target lesion(s) or/and maintenance of tumor marker level above the normal limits

[0446] Progressive Disease (PD):

[0447] Appearance of one or more new lesions and/or unequivocal progression of existing non-target lesions

Evaluation of Best Overall Response

[0448] The best overall response is the best response recorded from the start of the treatment until disease progression/recurrence (taking as reference for PD the smallest measurements recorded since the treatment started). In general, the patient's best response assignment will depend on the achievement of both measurement and confirmation criteria

[0449] Patients with a global deterioration of health status requiring discontinuation of treatment without objective evidence of disease progression at that time should be classified as having "symptomatic deterioration". Every effort should be made to document the objective progression even after discontinuation of treatment.

[0450] In some circumstances it may be difficult to distinguish residual disease from normal tissue. When the evaluation of complete response depends on this determination, it is recommended that the residual lesion be investigated (fine needle aspirate/biopsy) to confirm the complete response status.

Results

Patient Disposition

[0451] Disposition of patients is summarized herein: 26 subjects in 6 cohorts were enrolled between November 2007 and August 2009, and a total of 33 subjects had enrolled by December 2010. Each of the 3 subjects in cohorts 1-4 discontinued the study prematurely: 2 subjects due to death, 8 due to voluntary subject drop out, and 2 to pursue other treatment protocols. Of the 2 deaths, patient no. 1 was reported as a serious adverse effect, whereas patient no. 9 died 88 days post dosing and therefore, as defined in the protocol, was not reported as an SAE. In cohort 5, one subject completed the study (up to day 56), one withdrew prior to day 56 and one designated as "Other" withdrew due to disease progression. Five cohort 6 patients completed the study, 5 withdrew due to disease progression (2 had brain metastases), and one died approximately 2 months after receiving study treatment. Four cohort 7 patients completed the study; two withdrew due to disease progression.

[0452] Among the initial 33 patients enrolled, 31 attended the day 28 visit, and 19 attended the day 56 visit. Subjects were contacted for follow-up every 2-3 months thereafter for a period of up to 3 years or more post dosing for monitoring.

Patient Population Profile

[0453] Demographics

[0454] Demographics and baseline characteristics are summarized in Table 1 that follows. Overall, 54% of the subjects were male and 46% female. All but five of the subjects (4-Hispanic and 1-Asian) enrolled were Caucasian. The mean age of the subjects enrolled was 58.5 years, with the youngest 35.1 years old and the oldest 74.1 years. The mean Karnofsky score at entry was 85.4; the lowest score at entry was 70 (the minimum permitted by the protocol inclusion criteria) and the highest 100.

TABLE-US-00003 TABLE 1 Demographics/Baseline Characteristics Summary Demographic/Baseline Characteristic Number % Gender Male 14 53.8 Female 12 46.2 Race Caucasian 21 80.8 Asian 1 3.8 Hispanic 4 15.4 Age, mean (years) 58.5 -- Karnofsky score (mean) 85.4 --

[0455] Medical History

[0456] Medical history is summarized herewith: As expected in a patient population with a mean age of 58.5 years, all subjects had other medical conditions at baseline. The most common medical conditions (defined as those present in at least three subjects) were: hypertension (13 subjects); fatigue (10 subjects); constipation (6 subjects); diarrhea and anemia (5 subjects each); nausea, GERD, hypercholesterolemia, hyperlipidemia, seasonal allergy, and cholecystectomy (4 subjects each); abdominal pain, cough, hypothyroidism, low back pain, and pneumonia (3 subjects each).

[0457] Primary Diagnosis

[0458] The frequency of tumor type at baseline is summarized in Table 2 that follows. The most frequent tumor types were colorectal adenocarcinoma, non-small cell lung cancer, melanoma, sarcoma, and carcinoid/neuroendocrine. One subject each had the following tumor types: transitional cell carcinoma of the bladder, cancer of the distal esophagus, Merkle cell carcinoma, lung small cell carcinoma, renal cell carcinoma, gastro-intestinal stromal tumor, testicular sex cord tumor, and papillary thyroid carcinoma.

TABLE-US-00004 TABLE 2 Primary Tumor Diagnoses (cohorts 1-7) Number at Tumor Type Total number cohort 6-7 Colorectal Adenocarcinoma 11 5 Carcinoid/Neuroendocrine 4 4 Non-small cell lung cancer 3 1 Renal Cell Carcinoma 3 3 Melanoma 2 1 Sarcoma 2 0 Thyroid cancer 2 2 Esophageal Adenocarcinoma 1 1 Merkle cell carcinoma 1 1 Bladder/Transitional Cell 1 0 Lung small cell carcinoma 1 0 G-I stromal tumor 1 0 Testicular/sex cord tumor 1 0

Prior Chemotherapy

[0459] Prior chemotherapy is summarized by the number and frequency of chemotherapy herewith: All subjects enrolled in the trial had previous chemotherapy with multiple agents, with 3.7 (range 1-8) mean previous chemotherapy lines. The most frequently utilized medications were: 5-FU (10.9%), Bevacizumab (10.9%), Capecitabine (8.6%), Leucovorin (8.0%), Irinotecan (8.0%), Cetuximab (5.2%), Oxaliplatin (4.6%), Temozolomide (3.4%), Cisplatin (3.4%), Folfox (3.4%), Sunitinib (2.9%), Dacarbazine (2.3%), and Docetaxel (2.3%). Most subjects (16/26, 61.5%), received at least one prior anti-angiogenic agent: Bevacizumab (12 patients), Sunitinib (4 patients), and Sorafenib (2 patients). The use of these chemotherapeutic agents reflects the types of tumors occurring in this patient population.

[0460] Other Anti-Tumor Interventions

[0461] Essentially all subjects had previous surgery for their tumor. The details of the surgery and of other anti-tumor interventions, excluding chemotherapy, are summarized herewith: The average number of previous surgical procedures related to the tumor was >2/subject, with a few subjects having had 4 surgical procedures. Also, 12 of the subjects had previous radiation therapy. All of the courses of radiation therapy occurred at least 8 months prior to study treatment except for subject no. 13, whose last course of radiotherapy ended about 4.5 months previously. The protocol required no radiotherapy for at least 4 weeks prior to enrolment.

[0462] Tumor Evaluation at Baseline

[0463] All subjects had CT evaluations of tumor target lesions with measurement of tumor size at baseline for subsequent comparison post-treatment. For these measurements, the longest diameter of each target lesion was utilized.

[0464] Summary of Patient Population

[0465] Most subjects enrolled in this study were Caucasian with an almost equal gender distribution and a mean age of 58.5 years. The mean entry Karnofsky score was 85.4, and all subjects had concurrent medical conditions. The most frequent tumor types were colorectal adenocarcinoma, non-small cell lung cancer, melanoma, sarcoma and carcinoid/neuroendocrine. All subjects enrolled in the trial had previous cancer-related surgery and chemotherapy with multiple agents, the most common of which were 5-FU, Bevacizumab, Capecitabine, Leucovorin, Irinotecan, Cetuximab, Oxaliplatin, Temozolomide, Cisplatin, Folofox, Sunitinib, Dacarbazine, and Docetaxel; the majority also had previous anti-angiogenic therapy and radiation therapy.

Safety Evaluations

[0466] Adverse Events

[0467] The incidence and frequency of adverse events are summarized in Table 3 that follows: Overall, events experienced by at least 4 (15.4%) subjects included: pyrexia (50%); fatigue (46.2%); prolonged aPTT (38.5%); chills, hyponatremia and decreased hemoglobin (26.9% each); constipation (23.1%); nausea, vomiting, anorexia, and dyspoena (19.2% each); lymphopenia, elevated aspartate aminotransferase, hyperglycemia, myalgia, and hyperhidrosis (15.4% each). Pyrexia and chills occurred at higher doses only (cohorts 4-6). In cohort 6, 9 of 11 subjects experienced pyrexia and 6 of 11 also had chills. In cohort 7, one subject experienced a grade 3 fever. In all patients, the pyrexia and chills occurred on the day of study treatment (post-infusion), were transient, and resolved within 24 hours.

TABLE-US-00005 TABLE 3 Most Frequent Adverse Events (cohort 1-6) Adverse Event Number Patients % Pyrexia 13 50.0 Fatigue 12 46.2 aPTT Prolonged 10 38.5 Chills 7 26.9 Hyponatremia 7 26.9 Hemoglobin decreased 7 26.9 Constipation 6 23.1 Nausea 5 19.2 Vomiting 5 19.2 Anorexia 5 19.2 Dyspnea 5 19.2 Lymphopenia 4 15.4 AST Increased 4 15.4 Hyperglycemia 4 15.4 Myalgia 4 15.4 Hyperhidrosis 4 15.4

[0468] Most of the events of pyrexia (11 of 14) and chills (7 of 8) that occurred in the higher dose groups were considered possibly, probably or definitely related to study drug. The only other events considered definitely related to study drug included two events of nausea and one event each of anemia, vomiting, fatigue, hyperhidrosis, anorexia, and headache. None of these events were classified as NCI Grade 3 or higher. Thus, no dose limiting toxicities had occurred. Most of the other adverse events were considered unrelated or unlikely related to study drug.

[0469] Of the most common adverse events, fatigue was considered unrelated in 9/12 subjects and unlikely in one subject; prolonged aPTT unrelated in 7/10 subjects; decreased hemoglobin was considered unrelated in all 7 subjects; and constipation unrelated in all occurrences. Note: The local investigator commented that the aPTT local lab test is highly sensitive (as used for screening for lupus anti-coagulant); indeed, all aPTT elevation adverse events occurred in patients of that clinic.

[0470] The following Grade 3 or higher adverse events were reported; all were considered unrelated or unlikely related to the investigational drug: Subject no. 001 (cohort 1): Grade 3 disease progression; subject no. 007 (cohort 3): Grade 3 hyponatremia and hyperkalemia; subject no. 008 (cohort 3): Grade 3 hemoglobin decreased; subject no. 009 (cohort 3): Grade 3 fatigue; subject no. 013 (cohort 4): Grade 3 hypokalemia, hyponatremia, hypochloremia, and fatigue; subject no. 025 (cohort 6): Grade 4 suicidal ideation; subject no. 027 (cohort 6): Grade 3 weakness; subject no. 033 (cohort 6): Grade 4 abdominal pain and Grade 3 hyperglycemia; subject no. 035 (cohort 6): Grade 3 elevated total bilirubin, elevated AST, elevated ALT, and elevated alkaline phosphatase; and subject no. 036 (cohort 6): Grade 3 hyponatremia. Disease progression in subject no. 1 was a serious adverse event; all of the other events were not serious. All of the subjects with Grade 3 hyponatremia (no. 007, no. 013, and no. 036) with or without other Grade 3 metabolic events had gastro-intestinal malignancies.

[0471] Serious Adverse Events

[0472] No serious adverse events (SAEs) related to treatment with the study drug were reported.

[0473] Laboratory Testing:

[0474] Hematology and Coagulation

[0475] No significant trends in mean changes occurred after treatment in any of the cohorts for the following tests: basophil count, eosinophil count, lymphocyte count, hematocrit, hemoglobin, neutrophil count, platelet count, RBC, and PT. A trend for increased aPTT after treatment was observed in each cohort, although most mean values remained within the normal range. Mean and median white blood cell counts (WBC) tended to decrease in cohorts 3-6 from the day of dosing to day 4, but individual values remained within normal limits in all subjects except for two subjects in cohort 6: (1) subject no. 26 had a low WBC (3700 K/μL) on the day of dosing that decreased to 2700 K/μL on Day 4 but then increased to at least 4000 K/μL for the remainder of the study and (2) subject no. 32 had a normal WBC of 6.3 K/μL on the day of dosing that decreased to 4.5 K/μL on Day 4 and then varied between 4.3 and 6.0 K/μL.

[0476] Values for hematology tests that were considered clinically significant findings include:

[0477] Subject no. 8 (in cohort 3) had a hematocrit of 32.3% and hemoglobin of 10.6 g/dL on the day of dosing that gradually decreased over time to 25.5% and 7.9 g/dL, respectively, on day 28. This subject's platelet count was 443,000 K/μL on the day of dosing and increased to 516,000 K/μL on day 28.

[0478] Subject no. 14 (in cohort 4) had a hematocrit of 34.2% and hemoglobin of 10.8 g/dL at screening; these values gradually decreased over time to 26.4% and 8.3 g/dL, respectively, on day 14. This subject's aPTT was 33.1 seconds at screening, 34.4 seconds at day 4, and then increased to 49.6 seconds at day 7 and 53.6 seconds at day 14.

[0479] Subject no. 33 (in cohort 6) had a hematocrit of 38.3% at screening and 33.1% on the day of dosing and hemoglobin of 12.3 g/dL at screening and 10.9 g/dL on the day of dosing; both stabilized for the duration of the study. This subject's platelet count was 116,000 K/μL on the day of dosing and decreased to 89,000 MIL on day 4 but subsequently increased to 129,000 K/μL on Day 28. The PTT was normal at 31 seconds on the day of dosing but increased to 44.5 and 48.2 seconds on Days 14 and 28, respectively.

[0480] Summarizing the shift in hematology and coagulation lab values from the day of dosing to day 28 for the combined cohorts: The shift did not suggest any trend for values to shift to below or above the normal range at 28 days post-treatment. In particular, for both hematocrit and hemoglobin, no trend was observed for values to shift to below the normal range.

[0481] Chemistry

[0482] Summary of serum chemistry testing: No significant trends in median changes occurred after treatment in any of the cohorts for the following tests: ALT, AST, albumin, alkaline phosphatase, calcium, creatinine, glucose, potassium, sodium, bilirubin, total protein, and BUN. Mean levels of the following were increased, due to a high level observed in one subject: ALT and AST in cohort 6 at Day 28 due to levels of 406 U/L and 232 U/L, respectively, in subject no. 35; total bilirubin in cohort 6 at Day 28 due to a level of 7.4 in subject no. 35. Very high alkaline phosphatase levels in the following subjects accounted for elevated mean levels in their respective cohorts: subjects no. 4 (cohort 2), no. 24 (cohort 5), and no. 22, 26, 30, 35, and 36 (all in cohort 6).

[0483] Chemistry tests: Subjects no. 4 and no. 22 had elevated ALT and AST tests on the day of dosing that subsequently decreased/normalized. Subject no. 35 in cohort 6 had elevated levels of ALT and AST at day 28: 406 U/L and 232 U/L, respectively, but these levels normalized by day 56. This subject (no. 35) was hospitalized for bile duct obstruction due to tumor progression and also had markedly elevated levels of total bilirubin, 7.4 mg/dL, and alkaline phosphatase, 1176 U/L, at day 28. Otherwise, all ALT and AST elevations were <3X the upper limits of normal and tended to be sporadic. No subject other than no. 35 had an abnormal total bilirubin level during participation in the study. Elevated alkaline phosphatase on the day of dosing was common, presumably due to tumor and/or metastatic disease, but a significant progressive increase in this lab test only occurred in two subjects (no. 4 who had a level of 383 U/L at screening, 493 U/L on the day of dosing and 609 U/L at Day 28 and no. 36 who had a level of 163 U/L on the day of dosing and 377 U/L on Day 28). No subjects developed clinically significant post-treatment levels for any of the other chemistry tests.

[0484] Summarizing the shift in serum chemistry values from the day of dosing to day 28 for the combined cohorts: 5/15 subjects who had a normal level at the day of dosing had a low level of serum sodium on day 28. However, the sodium levels were abnormal only intermittently, usually only slightly below normal, and not clinically significant. The shift tables did not suggest any other trend for values to shift to below or above the normal range at 28 days post-treatment.

[0485] Urinalysis

[0486] The frequency of an abnormality on urinalysis on the day of dosing (89.5%) and at screening (76.0%) was high, but no trends were observed at the post-dosing visits. A review of the data indicated that no clinically significant abnormalities occurred on urinalysis, although a few subjects had small amounts of protein in their urine, and subject no. 30 in cohort 6 had 2+ proteinuria at day 28.

[0487] Antibodies to Adenovirus 5

[0488] Serum samples from subjects tested for antibodies to Adenovirus 5 (total and IgG); increase in titers between pre- and post-dose for IgG antibodies are summarized in Table 4 and FIG. 11. The post-dose samples were collected at day 28 or 56 in all but 3 subjects; these 3 had post-dose samples tested earlier due to the non-availability of samples at day 28. All post-dose IgG antibody titers increased at least 7-fold; 8 of 33 subjects had at least a 100-fold increase in IgG antibodies over the pre-dose titer. All post-dose total antibody titers to adenovirus 5 increased at least 5-fold; 10 of 26 subjects had at least a 625-fold increase in total antibodies over the pre-dose titer. There was no correlation between fold increase (total and IgG antibody titers) and dose level (P>0.05, Pearson and Spearman correlation tests). Data collected at day 56 for cohort 7 subjects indicated a trend of decreased IgG levels, compared to day 28 (see Table 4 that follows).

TABLE-US-00006 TABLE 4 Summary of Anti-Adenovirus 5 Increase in IgG Antibody Titers and Total Antibody Titers Anti-Ad5 Titer Anti-Ad5 IgG Titer Post-dose Fold Pre- Post-dose Fold Post-dose Cohort Patient Pre-dose (day 28) increase dose (day 28) increase (day 56) 1 1 62500 1562500 25 3500 43,740 12 2 2500 1562500 625 400 43,740 109 3 12500 1562500 125 1500 43,740 29 2 4 2500 62500 25 300 5,500 18 5 500 12500 25 180 1500 8 6 2500 7812500 3125 350 35,000 100 3 7 2500 12500 5 600 4100 7 8 2500 312500 125 950 43,740 46 9 2500 7812500 3125 1000 29,000 29 4 10 12500 1562500 125 420 20,000 48 13 2500 1562500 625 200 9,000 45 14 2500 1562500 625 650 11,000 17 5 20 12500 312500 25 90 13,000 144 21 2500 312500 125 300 14,000 47 24 12500 312500 25 1,200 12,000 10 6 22 12500 1562500 125 850 11,500 14 25 2500 312500 125 850 11,000 13 26 2500 312500 125 70 11,000 157 27 12500 7812500 625 540 43,740 81 28 ND 312500 ND 700 43,740 62 29 12500 312500 25 120 18,000 150 30 62500 7812500 125 6000 43,740 7 32 2500 1562500 625 200 43,740 219 33 2500 7812500 3125 500 43,740 87 35 2500 62500 25 ND ND ND 36 2500 7812500 3125 400 30,000 75 38 12500 7812500 625 800 43,470 54 7 39 62,500 1,562,500 25 800 30000 38 18000 40 12,500 312,500 25 300 35000 117 22000 41 62,500 7,812,500 125 200 7500 38 10000 42 12,500 1,562,500 125 450 5500 12 4500 43 62,500 1,562,500 25 140 7000 50 5000 44 12,500 312,500 25 200 22000 110 NA

[0489] All subjects were also assessed as to their level of neutralizing Adenovirus 5 antibodies at baseline (Table 5). Results show that 35% had highly elevated baseline levels (>210) and 41% of the subjects had low levels (≦18) of neutralizing antibodies.

TABLE-US-00007 TABLE 5 Summary of Anti-Adenovirus 5 Neutralizing Antibody Titers at Baseline Subject ID Cohort Titer 001 1 620 002 1 750 003 1 210 004 2 10.5 005 2 10.5 006 2 400 007 3 5 008 3 380 009 3 210 010 4 1050 013 4 860 014 4 5 020 5 18 021 5 5 024 5 520 022 6 110 025 6 210 026 6 5 027 6 5 028 6 940 029 6 5 030 6 1250 032 6 195 033 6 220 035 6 10.5 036 6 5 038 6 620 039 7 1000 040 7 5 041 7 15 042 7 5 043 7 160 044 7 85

[0490] No correlation could be discerned between the presence of neutralizing anti-Ad5 antibodies at baseline and disease progression at days 28 (FIG. 2) or 56 (FIGS. 3A, 3B Table 5) (Pearson Product Moment Correlation; P>0.050; Spearman Rank Order Correlation; P>0.050).

[0491] Cytokine Levels in Peripheral Blood in Cohort 6

[0492] Peripheral blood cytokine levels were measured in cohort 6 patients at baseline, and at the following times after dosing with AD5-PPE-1-3XD-FAS-CHIMERA-: 6 hours, 4 days, 7 days, 14 days, 28 days, and 56 days post infusion. Results are summarized in Table 6 that follows.

TABLE-US-00008 TABLE 6 Summary of Mean Peripheral Blood Cytokine Levels in Cohort 6 Baseline 6 h D4 D7 D14 D28 D56 Cytokine (n = 10) (n = 10) (n = 2) (n = 9) (n = 9) (n = 9) (n = 3) IL-6 10.38 1018.28 2.92 7.2 47.99 12.13 0 IL-8 21.85 181.68 42.6 33.93 43.68 31.59 21.62 VEGF 73.47 54.82 0 147.09 132.77 133.53 0 FGF 11.92 9.95 5.75 20.42 19.55 12.43 7.30 TNF-a 0 1.53 0 0 0 0 0 sTNFRI 1746.44 3143.89 1725.60 2383.03 2706.33 2222.37 1456.77 sTNFRII 5819.81 7509.75 10063.71 7921.62 10155.62 7717.02 5866.12

[0493] A significant increase in mean IL-6 levels occurred 6 hours post-infusion; this level returned to baseline by day 4. A smaller increase in TNFR11 levels was noted, peaking between days 4 and 14. No major elevations occurred with the other measured cytokines.

[0494] Vital Signs

[0495] No significant post-treatment trends were observed for systolic BP, diastolic BP, and respiratory rate, and no subject developed any clinically significant abnormalities.

[0496] Significant elevations in temperature (>38 degrees Centigrade) occurred in 5 subjects at 6 hours post-infusion: one subject in cohort 5: no. 20 (38.3 degrees Centigrade) and 4 subjects in cohort 6: no. 26 (39.1 degrees Centigrade), no. 27 (39.7 degrees Centigrade), no. 32 (39.8 degrees Centigrade), and no. 36 (39 degrees Centigrade). In each, the temperature had normalized prior to or by day 4 (which was the first recording of temperature following that of 6 hours post-infusion). The mean heart rate was increased to 101.2 beats/minute in cohort 6 at 6 hours post-dosing. This was due to amore rapid heart rate in the subjects that had experienced pyrexia in this cohort (heart rate of 107-140 beats/minute). Weight is summarized herein: no major change in weight occurred, except for a decrease in mean weight at follow-up (75.0 kg at screening, 75.0 kg at day 56, and 63.5 kg at follow-up).

[0497] Physical Exams

[0498] No treatment-related or clinically significant changes on exams are apparent from the data of by-subject physical exams at screening and day 56.

[0499] ECGs

[0500] ECG parameters at screening are summarized herein: At screening, 15 subjects had normal ECGs, and 11 had abnormalities that were considered not clinically significant. The original protocol specified that a follow-up ECG should be performed at the day 56 follow-up visit. As most subjects withdrew from the study prior to day 56, in cohorts 1-5 only subject no. 7 had a follow-up ECG and this showed ECG changes considered not clinically significant.

[0501] The protocol was later amended to obtain the follow-up ECG at the day 28 visit. In cohort 6, there were 4 follow-up ECGs obtained: two were normal and 2 had minor, non-clinically significant findings with no major changes from the screening ECG.

[0502] Biodistribution of Ad5-PPE-1-3X-fas-chimera

[0503] Due to an error, no whole blood samples were drawn for some of the patients in cohorts 5 and 6[3]). Urine samples were tested for 11 of the 12 cohort 6 patients and for the 12 additional patients from lower cohorts tested for levels in blood.

[0504] Analysis of the Urine Samples for Presence of Adenovirus:

[0505] Maximum levels of Adenovirus Vector DNA detected in urine are summarized in Table 7.

TABLE-US-00009 TABLE 7 Maximum Levels of Adenovirus Vector in Urine (cohorts 1-6) Adenovirus Vector Cohort Subject ID DNA copies/μg gDNA 1 001 0 1 002 3.2 × 10³ (Day 28) 1 003 5.3 (Day 56) 2 004 6.2 × 10² (3 hr) 2 005 0 2 006 0 3 007 0 3 008 0 3 009 2.2 × 10³ (Day 4) 4 010 5.3 (Day 14) 4 014 1.1 × 10³ (6 hr) 5 024 5.3 (6 hr) 6 022 1.1 × 10⁴ (3 hr) 6 025 1.5 × 10⁴ (3 hr) 6 026 5.3 (3 hr) 6 028 5.3 (3 hr) 6 029 1.6 × 10⁷ (3 hr) 6 030 4.7 × 10⁵ (3 hr) 6 032 5.3 (3 hr) 6 033 2.3 × 10⁴ (3 hr) 6 035 1.8 × 10⁴ (3 hr) 6 036 0 6 038 0 VP--Viral particles; BLQ < 20 copies; BLD < 1.4 copies

[0506] Analysis of the Whole Blood Samples for the Presence of Adenovirus Vector

[0507] Average levels of Adenovirus Vector DNA, as detected by RT-PCR in whole blood for cohorts 1-6 are summarized over time in FIG. 12A. Table 8 that follows shows the median values for cohorts 1-6. Before infusion of the adenovirus vector, none of the patients tested showed amplification of adenoviral gene (below detectable levels). A dose-dependent increase in average maximum levels of adenovirus vector DNA found in whole blood is evident from the data. At the end of the infusion all patients had individual blood virus levels in the range of 1.9×10³ and 5.5×10⁷ copies/μg gDNA, correlating positively with dose received.

[0508] In cohort 6 patients (FIG. 12A, grey dashed line), average levels of adenovirus decreased from a range of 2.1×10⁵-5.5×10⁷ (end of infusion) to a range of 1.1×10⁴-2.6×10⁵ copies/μg gDNA (three hours post infusion, see FIG. 12B). Levels of adenovirus continued to decrease 6 hours post infusion and subsequently decreased throughout the final time points. By day 56, Adenovirus DNA levels were either all reduced by at least 2 log-fold, or were undetectable.

TABLE-US-00010 TABLE 8 Presence of Adenovirus Vector in Whole Blood: Median Values Median Adenovirus Vector DNA copies/μg gDNA at end of infusion Cohort Dose vp (Maximum) 1 1 × 10¹⁰ 1.6 × 10⁴ 2 3 × 10¹⁰ 4.5 × 10⁵ 3 1 × 10¹¹ 1.6 × 10⁵ 4 3 × 10¹¹ 1.7 × 10⁶ 5 1 × 10¹² 6.5 × 10⁶ 6 3 × 10¹² 2.5 × 10⁷

[0509] Analysis of the Expression Levels of Fas-Chimera Transgene in Whole Blood

[0510] None of the 21 subjects tested had detectable levels of transgene cDNA (as determined by RT-PCR, representing blood mRNA levels) in whole blood. However, in one patient 01-036 (esophageal cancer) a sample from a subcutaneous metastasis was tested and detectable levels of adenovirus transgene expression were found in the aspirate on days 4 (1.4×10⁵ copies/μg RNA) and 28 (3.9×10⁵ copies/μg RNA) after treatment (Table 9), providing direct evidence of the specificity of the transgene expression in the target tumor tissue.

TABLE-US-00011 TABLE 9 Aspirate from tumor of patient 01-036 (copies/μg RNA) time Pt no. cohort sample copies Pre 36 6 Tumor 0.00E+00 Day 4 36 6 Tumor 1.40E+05 Day 28 36 6 Tumor 3.90E+04

[0511] Summary of Safety Results

[0512] No signs of significant safety issues with Ad5-PPE-1-3X-fas-chimera were observed in this study. The maximal therapeutic dose (MTD) was determined to be 10³ virus particles per dose, in view of a dose limiting toxicity of a NCI grade 3 fever, shortly following dosing at the highest dose tested (cohort 7). Other than abnormal lab results, the most frequent adverse events were pyrexia, fatigue, chills, and constipation. Pyrexia and chills occurred mostly in the higher dose groups and were usually considered related to study medication. Although no trend for hematology values to shift outside the normal range was observed, several subjects had declines in hemoglobin and hematocrit values after study treatment. No clinically significant post-treatment chemistry or urinalysis abnormalities occurred during the study with the exception of a markedly elevated alkaline phosphatase and elevations of ALT, AST, and total bilirubin occurring in a subject with bile duct obstruction due to progression of cancer. No clinically significant post-treatment changes in vital signs were observed, except for the occurrence of fever 6 hours post-infusion in one cohort 5 and 4 cohort 6 subjects; the cohort 6 subjects with fever also had increased heart rates, resulting in a mean increase in heart rate for cohort 6 at 6 hours post-infusion. No treatment-related changes were observed on physical exams or ECGs.

[0513] All subjects had pre-dose anti-Adenovirus 5 antibodies (total and IgG) with total antibody titers increasing at least 5-fold post-treatment. FIG. 11 shows the anti-Adenovirus 5 antibodies in each of cohorts 1-6, measured at baseline, day 7, day 14 and day 28, and illustrates the tendency of the anti-Adenovirus 5 antibodies to plateau is between 7 and 14 days post infusion, despite the variability observed between patients in the baseline antibody titer levels. Levels of anti-Adenovirus 5 antibodies (total and IgG) tended to increase post dosing, peaking on day 28 and tending to decrease by day 56 (FIG. 11). Overall, the fold increase was higher for the total anti-Adenovirus 5 titers than for the IgG titers, but no correlation was discerned between fold increase in anti-Adenovirus 5 antibodies and the dosage level. 34.6% of subjects had highly elevated levels of neutralizing antibodies to Adenovirus 5, measured at baseline, 23.1% had moderately elevated levels, and 42.3% had low levels. However, there was no correlation discernable between neutralizing anti-body titer at baseline and a clinical response (measured as stable disease). A significant increase in mean IL-6 blood levels occurred 6 hours post-dosing with Ad5-PPE-1-3X-fas-chimera; this level returned to baseline by day 4. No major elevations occurred with the other measured cytokines (II-8, VEGF, FGF, and TNF-alpha).

[0514] In most subjects with detectable levels of Adenovirus 5 in the urine, the presence was transient, with levels detectable only within the initial 24 hours after the intravenous (IV) infusion of Ad5-PPE-1-3X-fas-chimera. The Ad5-PPE-1-3X-fas-chimera adenovirus vector was present in high copy numbers in whole blood directly after the IV infusion. The levels of adenovirus vector subsequently decreased with time in whole blood. Samples with vector present in the whole blood were tested for the expression of the Fas-chimera transgene (RT-PCR for the transgene mRNA). None of the 21 subjects tested had detectable levels of transgene cDNA (cDNA is the RT-PCR reaction product representing blood mRNA levels) in whole blood.

Preliminary Efficacy Assessment

[0515] Disease Progression or Recurrence

[0516] Disease progression was assessed according to clinical deterioration and radiographic growth, based on RECIST criteria. In cohorts 1-5, at day 56, 3/14 patients had stable disease. In cohort 6, on day 56 (n=12), 1 PR, 4 SD ("stable disease") and 7 had progressive disease. Thus at day 56, 5/12 (42%) had stable disease or better. In cohort 7, at day 56, 4 out of 6 subjects (67%) had stable disease: In cohorts 1-6, at day 28, 21/26 (80.8%) of the subjects were considered not to have deterioration, one had deteriorated (5.6%), and 4 had no observation for this endpoint. Only 8 subjects from cohorts 1-6 had observations at the Day 56 visit, and all of these were negative for deterioration.

[0517] Disease progression was also assessed by frequency of radiographic growth (measured according to % growth in the sum longest diameter of target lesions as defined in the RECIST scoring criteria). Where no new lesions were present and there was either no growth in the longest diameter sum of the target lesions or that growth was no more than 20%, the subject was considered to have stable radiographic disease at that visit.

[0518] Reduction in Refractory Metastatic Papillary Thyroid Cancer Following Multiple Infusions of 3×10¹² Virus Particles of Ad5-PPE-1-3X-fas-chimera

[0519] Of these patients, Subject 026, a 69 year-old Hispanic female, was enrolled with metastatic papillary thyroid cancer that was resistant to radioiodine and was dosed in cohort 6 on Mar. 16, 2009. Baseline CT scan showed a mass lesion in the neck with pressure on the airway (FIG. 4, arrows). Day 28 scan showed stable disease, and a follow-up scan 6 months (FIG. 5, arrows) and 12 months after treatment showed a greater than 30% reduction in the long diameter of the mass and no pressure on the airway, with radiographic lucency suggestive of central necrosis (FIG. 5, blue arrow). Additionally, thyroglobulin levels have decreased in this patient: the level at baseline was 426 ng/mL and 10 months later had decreased to 326 ng/mL (normal levels<55 mg/mL).

[0520] A second dose was administered one and one half years after the first dosing, and the patient was progression free at day 120 post infusion. Thyroglobulin levels at the time of second dosing were 281 ng/mL, rising to 477 ng/mL at day 28, and dropping to 382 ng/mL at day 56 post infusion. No dose limiting toxicities, severe side effects or cytokine storm were observed throughout the treatment period. (FIG. 10A)

[0521] In another patient with a medullary thyroid cancer lesion, administration of 3×10¹² virus particles of Ad5-PPE-1-3X-fas-chimera resulted in dramatic reduction in levels of the thyroid cancer biomarker calcitonin: levels were greatly elevated before administration (14, 331 pg/ml, normal=<10 pg/ml), increased (16,324 pg/ml) at 56 days post infusion, but showed marked reduction (6600 pg/ml) at 120 days post infusion. At 120 days post-treatment the patient was still progression free. (FIG. 10B)

[0522] Reduction in metastatic lesion of advanced neurorendocrine cancer following a single infusion of 3×10¹² virus particles of Ad5-PPE-1-3X-fas-chimera.

[0523] Another patient in cohort 6, enrolled with advanced neuroendocrine cancer, was dosed in cohort 6 on Mar. 16, 2009. Day 21 CT scan showed several hepatic metastases (FIGS. 6A and 7A, red circle). Follow-up CT scans at 50 (FIGS. 6B and 7B, red circle) and 112 (FIGS. 6C and 7C, red circle) days post dosing showed continued regression of one of these metastatic lesions. According to RECIST criteria, this patient was scored as "stable disease", and has maintained "stable disease" classification for four months afterwards.

[0524] Summary of Efficacy

[0525] Although this single infusion study with patients having advanced or metastatic solid organ cancer is small, certain trends can be discerned from the efficacy data.

[0526] On day 56 evaluation, three of the 14 patients in Cohorts 1-5 had stable radiographic disease (SD); among the 12 Cohort-6 patients, five had stable disease on day 56, and one patient (with papillary thyroid carcinoma) had a near partial response on day 56 (out of 12 patients on cohort 6), which became a partial response (PR) later.

[0527] Among the 5 cohort 7 patients, 5 remained classified as having "stable disease" (SD). When the percent change in RECIST scores is plotted for days 0, 28 and 56, and correlated with the evaluations of stable and progressive disease, it can be seen that three out of 14 (21%) in cohorts 1-5 were stable at day 56 (FIG. 8). In cohorts 6 and 7 (FIG. 9), however, 9 out of 17 patients were stable at day 56.

[0528] In this open-label, dose-ranging study evaluating a single IV infusion of safety and efficacy of administration of Ad5PPE-1-3X-fas-chimera (SEQ ID NO: 9) in the clinical setting, 3 subjects with advanced or metastatic cancer have been enrolled in each of the first 5 ascending dose cohorts and 12 subjects have been enrolled in the 6^th cohort. Taken together, the data indicates Ad5-PPE-1-3X-fas-chimera is safe for systemic administration, in all of the doses tested, as no safety or dose-limiting toxicities (DLT) have been observed at any of the utilized doses. Although two serious adverse events were reported, these were related to progression of the cancer and unrelated to study treatment. Several subjects who received Ad5-PPE-1-3X-fas-chimera at one of the 2 highest doses developed transient pyrexia and chills shortly after receiving study treatment, events commonly occurring after the administration of adenovirus vectors. Clinically significant abnormalities on laboratory testing were infrequent and considered unrelated to Ad5-PPE-1-3X-fas-chimera treatment.

[0529] Maximally tolerated dose (MTD) was determined as 10¹³ virus particles per dose, in view of the single incident of dose limiting toxicity (NCI Grade 3 fever) observed in the cohort receiving the highest dose tested (cohort 7). Although being a small single infusion study among patients with multiple types of advanced, refractory cancer, evidence for efficacy of Ad5-PPE-1-3X-fas-chimera infusion includes patients with stable radiographic disease and one prolonged partial response among patients receiving 3×10¹² virus particles of Ad5-PPE-1-3X-fas-chimera (cohort 6) (Table 10).

TABLE-US-00012 TABLE 10 Disease response to a single dose of Ad5PPE-1-3X-Fas-chimera: Percent stable disease at day 56, by indication (cohorts 6 and 7). Number of % SD at Type of Cancer Patients Day 56 Colorectal Adenocarcinoma 5 40% Carcinoid/Neuroendocrine 4 75% Non-small cell lung cancer 1 100% Renal cell carcinoma 3 33% Melanoma 1 0% Thyroid cancer 2 100% Merkle cell carcinoma 1 0% Esophageal Adenocarcinoma 1 0%

[0530] In one patient suffering from esophageal cancer the subcutaneous metastasis was sampled and detectable levels of Ad5-PPE-1-3X-fas-chimera transgene expression were found in the aspirate on days 4 and 28 after treatment.

Example III

Effect of Ad5PPE-1-3X-fas chimera Administration on Thyroid Cancer in the Clinical Setting

[0531] In order to determine the efficacy of administration of AD5PPE-1-3X-fas-chimera on Thyroid Cancer in the clinical setting, outcomes such as toxicity, adverse effects, antibody titer, biodistribution, disease progression and disease recurrence and survival were monitored in subjects with Thyroid Cancer tumors receiving intravenous infusion of the Ad5PPE-1-3X-fas chimera adenovirus vector.

Materials and Experimental Methods

[0532] Indications:

[0533] Advanced progressive and radioiodine-refractory differentiated thyroid cancer (DTC) (papillary, follicular, Hurthle cell) cancer patients.

[0534] Safety Objectives:

[0535] To evaluate the safety of a single systemic dose of the Ad5PPE-1-3X-fas chimera adenovirus vector in patients with advanced thyroid cancer.

[0536] Efficacy and Pharmacodynamics Objectives:

[0537] 1. To evaluate the response to treatment for patients with advanced DTC with measurable disease using Response Evaluation Criteria in Solid Tumors (RECIST Criteria);

[0538] 2. To assess the pharmacokinetic and pharmacodynamic profile of Ad5PPE-1-3X-fas chimera adenovirus vector;

[0539] 3. Evaluation of changes in candidate biomarkers in response to Ad5PPE-1-3X-fas chimera adenovirus vector treatment.

[0540] 4. To explore influences of pre-treatment tumor genetic alterations on response to Ad5PPE-1-3X-fas chimera adenovirus vector treatment using archival tumor materials.

[0541] Efficacy Endpoints:

[0542] 1. The primary efficacy endpoint is the proportion of patients who have achieved an objective response to the study agent (according to RECIST criteria).

[0543] 2. Secondary endpoints will include changes in thyroglobulin levels in response to treatment (in anti-thyroglobulin antibody-negative patients only).

[0544] Study Design:

[0545] Prospective, open-label, single dose study in 2 groups of patients (parallel enrolment):

[0546] Group A--Treatment of 12 evaluable patients with progressive thyroid cancer disease despite treatment with radioiodine, but naive to targeted anti-angiogenic therapies (e.g. tyrosine kinase inhibitors or anti-VEGF monoclonal antibodies). Subjects may also have had treatment with other cancer chemotherapy.

[0547] Group B--Treatment of 12 evaluable patients with progressive thyroid cancer despite prior treatment with radioiodine and with at least one anti-angiogenic therapy. Subjects may also have had treatment with other cancer chemotherapy.

[0548] Treatment Plan and Study Duration:

[0549] Ad5PPE-1-3X-fas chimera adenovirus vectors are administered as a single intravenous infusion of 3×10¹² vp (virus particles). The post-infusion efficacy follow up period will be until disease progression occurs. The post-treatment safety and efficacy evaluation visits will be every four weeks until week 12 or disease progression (whichever occurs later). Thereafter, restaging evaluations will occur every 2 months until at least one year after study enrolment or until progression (the earlier). Formal restaging of indicator lesions is performed every 8 weeks.

[0550] Population Size:

[0551] Groups A and B of this study will each enroll 12 evaluable subjects, for a total of 24 evaluable subjects. Evaluable subject are subjects for whom the chosen evaluation criteria can be applied through the duration of the study. The trial is designed according to the 2-stage Simon statistical method. If at least 1 response is observed in the initial 12 patients, further 24 patients will be enrolled from that group (up to a total of 37 patients per group or 74 total).

[0552] Inclusion Criteria:

[0553] 1. Patients≧18 years of age;

[0554] 2. Histologically or cytologically confirmed advanced DTC (papillary, follicular, Hurthle cell);

[0555] 3. Absence of sensitivity to therapeutic radioiodine;

[0556] 4. No previous treatment with anti-angiogenic agents (Group A patients only);

[0557] 5. Measurable disease, defined as at least one lesion that can be accurately measured in at least one dimension (longest diameter to be recorded) as ≧20 mm with conventional techniques or as ≧10 mm with spiral CT scan (Note: Disease that is measurable by physical examination only is not eligible, and CT and/or MRI are required in assessing indicator lesions);

[0558] 6. Life expectancy>3 months;

[0559] 7. ECOG performance status (PS) 0, 1, or 2; Karnofsky performance status of ≧60%;

[0560] 8. Objective evidence of tumor progression in the 6 month period prior to the Screening Visit, as assessed by:

[0561] i. Progressively increasing suitable tumor markers, where appropriate; and

[0562] ii. Unequivocal progression of objectively measured disease on successive appropriate imaging (e.g. CT scan). In cases of uncertainty of tumor progression, the Principal Investigator of the study will be available to assist in decisions.

[0563] 9. Patients with a normal/acceptable hematological profile, as demonstrated by a peripheral leukocyte count>3000 cells/mcL, an absolute neutrophil count>1500 cells/μl, hemoglobin≧10 g/dl, platelets>100,000/μl, and International Normalized Ratio (INR)<1.2× the Upper Limit of Normal (ULN);

[0564] 10. Patients with adequate renal function, i.e. serum creatinine<1.5 times upper normal limits; and adequate hepatic function, as defined by ALT and AST<2.5× the upper limit of normal and total bilirubin below the upper limit of normal;

[0565] 11. Males and females of childbearing potential must utilize, throughout the course of the trial a standard contraception method;

[0566] 12. Ability to understand and the willingness to sign a written informed consent document;

[0567] 13. Willingness to comply with study requirements.

[0568] Exclusion Criteria:

[0569] 1. Pregnant or breastfeeding females;

[0570] 2. Disease that is measurable by physical examination only;

[0571] 3. Presence of any of the following:

[0572] Radiotherapy or chemotherapy<4 weeks prior to baseline visit;

[0573] Radiotherapy to ≧25% of bone marrow;

[0574] Major surgery<4 weeks prior to baseline visit;

[0575] Concurrent and/or prior therapy with octreotide will be allowed, provided tumor progression on this therapy has been demonstrated;

[0576] Concurrent and/or prior therapy with biphosphonates will be allowed;

[0577] 4. Any other ongoing investigational agents within 4 weeks before enrolment;

[0578] 5. Patients, who suffered from an acute cardiac event within the last 12 months, including myocardial infarction, cardiac arrythmia, admission for unstable angina, cardiac angioplasty, or stenting;

[0579] 6. QTc prolongation (defined as QTc interval≧500 msecs) or other significant ECG abnormalities (e.g. frequent ventricularectopy, evidence of ongoing myocardial ischemia);

[0580] 7. Patients with active vascular disease, either myocardial or peripheral;

[0581] 8. Patients with proliferative and/or vascular retinopathy;

[0582] 9. Patients with known active liver disease (alcoholic, drug/toxin induced, genetic, or autoimmune) other than related to tumor metastases;

[0583] 10. Patients with known CNS metastatic disease (Exception: patients with treated CNS metastases stable by radiographic examinations>6 moths after definitive therapy administered, are eligible);

[0584] 11. Patients testing positive to one of the following viruses: HIV, HBV or HCV;

[0585] 12. Any of the following conditions:

[0586] Serious or non-healing wound, ulcer, or bone fracture;

[0587] History of abdominal fistula, gastro-intestinal perforation, active diverticulitis, intra-abdominal abscess or gastro-intestinal tract bleeding within 28 days of enrolment;

[0588] Any history of cerebrovascular accident (CVA) within 6 months of enrolment;

[0589] Current use of therapeutic warfarin (Note: Low molecular weight heparin and prophylactic low-dose warfarin [International Normalized Ratio (INR)<1.2× Upper Limit of Normal (ULN)] are permitted);

[0590] History of bleeding disorder, including patients with hemophilia, disseminated intravascular coagulation (DIC), or any other abnormality of coagulation potentially predisposing patients to bleeding;

[0591] Poorly controlled depression or anxiety disorder, or recent (within the previous 6 months) suicidal ideation;

[0592] 13. Patients with an ongoing requirement for an immunosuppressive treatment, including the use of glucocorticoids or cyclosporin, or with a history of chronic use of any such medication within the last 4 weeks before enrolment;

[0593] 14. Uncontrolled intercurrent illness including, but not limited to ongoing or active infection, or psychiatric illness/social situations that would limit compliance with study requirements.

[0594] Formulation, Dosage and Administration:

[0595] Ad5PPE-1-3X-fas chimera adenovirus vector is formulated as a sterile vector solution. The solution is supplied frozen (below-65° C.), in single use vials. Each vial contains 0.5 ml of vector solution at a specific viral titer. The vector solution is thawed and maintained on ice during dilution and handling.

[0596] AdSPPE-1-3X-fas chimera adenovirus vector is administered as a single intravenous infusion, maximal dose being 1×10¹²-1×10¹³ virus particles (vp). Prior to infusion, the solution for injection is brought to room temperature. The vials are opened in a biological safety cabinet and diluted (by injection) 1:4 with of normal saline for infusion. A single infusion of diluted Ad5PPE-1-3X-fas chimera adenovirus vector is administered at a rate of 1 ml/minute. Multiple doses may be administered at predetermined minimal intervals.

[0597] Safety Evaluation:

[0598] 1. Adverse events are recorded on an ongoing basis during. The events are followed until resolved, or until the study end, whichever comes sooner.

[0599] 2. Vital signs are recorded at screening, prior to dosing, 30, 60 minutes, 4 and 6 hours after dosing and at all patient visits.

[0600] 3. A physical examination is conducted in conjunction with each study visit.

[0601] 4. Safety laboratory assessment (vital signs, blood haematology and chemistry, urine analysis) are conducted at screening, prior to dosing, and at all patient visits, starting from day 4±1 until disease progression or Week 12, whichever occurs later.

[0602] 5. After disease progression or Week 12 (whichever occurs later), each patient is contacted for safety follow-up by telephone every 2 months until 1 year after study enrolment.

[0603] Biodistribution:

[0604] Blood and urine samples for evaluation of levels of adenovirus vector DNA and the fas transgene are collected for adenovirus vector DNA: at several time points post-infusion.

[0605] Evaluation of Efficacy: Tumor Response

[0606] The effect of Ad5PPE-1-3X-fas chimera adenovirus vector treatment on tumor response is evaluated by measuring the tumor according to RECIST criteria (see above) at screening and at subsequent visits until disease progression. Changes in tumor volume are evaluated and analyzed based on radiological studies.

[0607] Biochemical and Laboratory Assays:

[0608] Antibodies: Serum samples are collected prior to dosing and at all patient visits, starting from Week 1, for analysis of levels of antibodies to the adenovirus (both total immunoglobulin, total IgG and Ad-5 neutralizing Abs) and to the fas-chimera transgene.

[0609] Tumor markers: TSH, anti-thyroglobulin antibody, and thyroglobulin are tested in conjunction with all follow-up evaluations.

[0610] Tumor measurement: Evaluation of biological effect on indicator lesion (CT, MRI).

[0611] Statistical Evaluation:

[0612] Patients are divided into two groups of subjects based upon previous treatment of their thyroid cancer: Group A, DTC radioiodine resistant and naive to anti-angiogenics; Group B; DTC radioiodine resistant with progression on anti-angiogenics. These two groups are evaluated independently for efficacy and toxicity end points.

[0613] Patient recruitment for each group is in 2 stages:

[0614] Stage 1: Enter 12 patients into the study. 12 subjects for each group. If no clinical responses are observed, the therapy is considered ineffective in this patient population and the study is terminated. If at least 1 response is observed, the study group proceeds to Stage 2.

[0615] Stage 2: Enter an additional 25 evaluable patients into the each group study. If three or fewer responses are observed after all 37 evaluable patients have been evaluated for response, the therapy is considered insufficiently effective in this patient population. If 4 or more responses are observed, this is considered adequate evidence of promising activity and this treatment may be recommended for further testing in subsequent studies in this patient population.

[0616] Unless toxicity is encountered or the study is stopped at the interim analysis, this study will accruing 37 eligible subjects for each group and evaluable patients. In order to account for ineligibility, cancellation, major treatment violation, or other reasons or early withdrawal or drop out, an additional 4 patients will be enrolled for each group. Thus 41 patients will be enrolled into this study for each group. Total of 82 subjects.

[0617] Assessments will be carried out according to the schedule in Table 10 that follows.

TABLE-US-00013 TABLE 10 SCHEDULE OF ASSESSMENTS D₀ Prior to Follow up Week Weeks¹⁰ Screening Dosing after Week 1 Week 2 Weeks 12, 20, 28, 36, (≦2 wks day of Dosing Day ±1 ±1 4 & 8 ±3 44, 52 from D₀) dosing (D.sub.≦1) 4 ± 1 Day Day ±3 Days Days ±3 Days F/U¹¹ Inclusion Criteria X Exclusion Criteria X Medical History X Informed Consent X Pregnancy Test¹ X Physical Exam ECG X X Vital Signs² X X X X X X Hematology³ X X X X X X X X Chemistry⁴ X X X X X X X X¹² HIV, HBV, HCV X Urinalysis⁵ X X X X X X X ECG X X Drug infusion X Antibody⁶ X X X X X X X Distribution⁷ X X X X X X Concomitant Medications X X X X X X Adverse Events X X X X X X X X X Tumor Measure⁸ X X X XX Tumor Markers⁹ X X X XX Legend ¹Women of childbearing potential will be tested for pregnancy with the use of a serum pregnancy test at Screening and Week 12 ²Vital signs (blood pressure, body temperature and heart rate) will be recorded at screening, prior to dosing at 30 and 60 minutes after dosing and at 6 hours post-dosing, and at all patient visits thereafter. ³Hematology will include: complete blood count with differential, PT and PTT Fibrinogen ⁴Chemistry will include: electrolytes (sodium, potassium, calcium), creatinine; bilirubin, alkaline phosphatase, ALT and AST; total protein and albumin ⁵Routine urine analysis ⁶Blood samples will be collected for levels of serum antibodies against the adenovirus and the transgene ⁷Blood and urine samples will be collected for biodistribution determination (levels of the viral DNA and transgene). See protocol for details. ⁸Evaluation of indicator lesion (CT, MRI, etc.) to be done at each visit until disease progression ⁹TSH, anti-thyroglobulin antibody, and thyroglobulin should be tested at each visit until disease progression ¹⁰Visits at Weeks 20, 28, 36, 44, and 52 to be conducted only if no disease progression has occurred prior to the visit ¹¹Telephone contact will be performed every 2 months after Week 12 or disease progression (whichever occurs later) until one year after study enrolment.

[0618] Thyroglobulin is to be tested every 2 months until 13 months post dosing or until progression, whichever occurs first.

Example IV

Effect of Ad5PPE-1-3X-fas Chimera Administration Combined with Chemotherapy

[0619] In order to evaluate the combined therapeutic oncolytic effect of Ad5PPE-1-3X-fas chimera and chemotherapy on tumor size in cancer, systemic administration of Ad5PPE-1-3X-fas chimera and concomitant chemotherapy in the rapidly metastasizing Lewis Lung Carcinoma model was chosen.

[0620] The Lewis Lung Cancer model provides a method for observing the effects of treatment on established, metastatic cancer.

[0621] Sunitinib (Sutent) targets tyrosine kinase, and inhibits the action of VEGF, producing an anti-angiogenic effect, and is used, among others, in stromal tumors and advanced renal cell cancer.

[0622] Antimetastatic Effects of a Single Systemic Dose of Ad5PPE-1-3X-Fas Chimera and Oral Sunitinib in Mice Bearing Lung Metastases:

[0623] C57BL/6 male mice (13 to 19 in each group) aged 8 weeks were inoculated with 5×10⁵ LLC cells into the left footpad. The foot was amputated under general anesthesia as soon as the primary tumor developed to 7 mm. 2 days later (post foot amputation) a single intravenous injection of Ad5PPE-1-3X-fas chimera (10⁹ or 10¹¹ virus particles) was administered through the tail vein. After receiving the vector, a daily regimen of oral sunitinib was administered, 40 or 80 mg/kg once a day, on days 1-5, 8-13 and 16-17. Mouse sacrifice was scheduled for the 22^nd day post primary tumor removal. Mouse well-being was monitored daily by observation and weighing. Results (Tumor burden) relate to the tumor mass, in grams (known as Tumor Burden).

[0624] FIGS. 13A and 13B detail the results of two groups of the metastatic mice receiving the combination therapy, compared to each treatment mode alone. While a dose effect of the treatments could be discerned in the tumor burden of mice receiving 80 mg/kg sunitinib compared to those receiving 40 mg/kg sunitinib, and in the tumor burden of mice receiving 10¹¹ Ad5PPE-1-3X fas-c compared to those receiving 10⁹ Ad5PPE-1-3X fas-c, the results of combining the two treatment modes reveals a statistically significant difference (P<0.05) between the control group and all the treatment groups, the mean tumor burden in the control group being significantly greater than in each of the other groups. The high viral dose (10¹¹ virus particles) and the low dose-combination treatment (10⁹ virus particles and 80 mg/kg sunitinib), were found to be most effective in reducing the tumor burden, resulting in a statistically lower tumor burden than that of either the 10⁹ virus particles and 40 mg/kg sunitinib groups. These combination groups also showed reduced variability and generally lower scores, compared to the other experimental groups. The results show that combined treatment of systemically administered AdPPE-1-3X-fas-chimera+oral sunitinib is effective against metastatic disease.

[0625] Taken together, these results indicate that when administered in combination with currently employed clinical chemotherapy protocols, AdSPPE-1-3X fas-c can increase their therapeutic effectiveness and potentially allow reduced dosage and frequency of treatments.

[0626] Although the invention has been described in conjunction with specific embodiments thereof, it is evident that many alternatives, modifications and variations will be apparent to those skilled in the art. Accordingly, it is intended to embrace all such alternatives, modifications and variations that fall within the spirit and broad scope of the appended claims.

[0627] All publications, patents and patent applications mentioned in this specification are herein incorporated in their entirety by reference into the specification, to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated herein by reference. In addition, citation or identification of any reference in this application shall not be construed as an admission that such reference is available as prior art to the present invention. To the extent that section headings are used, they should not be construed as necessarily limiting.

Sequence CWU 1

1

644134350DNAArtificial sequenceEmpty Ad5 vector sequence without repeats 1catcatcaat aatatacctt attttggatt gaagccaata tgataatgag ggggtggagt 60ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg tagtagtgtg gcggaagtgt 120gatgttgcaa gtgtggcgga acacatgtaa gcgacggatg tggcaaaagt gacgtttttg 180gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg gttttaggcg gatgttgtag 240taaatttggg cgtaaccgag taagatttgg ccattttcgc gggaaaactg aataagagga 300agtgaaatct gaataatttt gtgttactca tagcgcgtaa tatttgtcta gggccgcggg 360gactttgacc gtttacgtgg agactcgccc aggtgttttt ctcaggtgtt ttccgcgttc 420cgggtcaaag ttggcgtttt attattatag tcagtacgtc tcgagcatgc atctaggcgg 480ccgcatggca gaaattcgcg aattcgctag cgttaacgga tcctctagac gagatccgaa 540cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 600taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 660tcatgtctag atctgtactg aaatgtgtgg gcgtggctta agggtgggaa agaatatata 720aggtgggggt cttatgtagt tttgtatctg ttttgcagca gccgccgccg ccatgagcac 780caactcgttt gatggaagca ttgtgagctc atatttgaca acgcgcatgc ccccatgggc 840cggggtgcgt cagaatgtga tgggctccag cattgatggt cgccccgtcc tgcccgcaaa 900ctctactacc ttgacctacg agaccgtgtc tggaacgccg ttggagactg cagcctccgc 960cgccgcttca gccgctgcag ccaccgcccg cgggattgtg actgactttg ctttcctgag 1020cccgcttgca agcagtgcag cttcccgttc atccgcccgc gatgacaagt tgacggctct 1080tttggcacaa ttggattctt tgacccggga acttaatgtc gtttctcagc agctgttgga 1140tctgcgccag caggtttctg ccctgaaggc ttcctcccct cccaatgcgg tttaaaacat 1200aaataaaaaa ccagactctg tttggatttg gatcaagcaa gtgtcttgct gtctttattt 1260aggggttttg cgcgcgcggt aggcccggga ccagcggtct cggtcgttga gggtcctgtg 1320tattttttcc aggacgtggt aaaggtgact ctggatgttc agatacatgg gcataagccc 1380gtctctgggg tggaggtagc accactgcag agcttcatgc tgcggggtgg tgttgtagat 1440gatccagtcg tagcaggagc gctgggcgtg gtgcctaaaa atgtctttca gtagcaagct 1500gattgccagg ggcaggccct tggtgtaagt gtttacaaag cggttaagct gggatgggtg 1560catacgtggg gatatgagat gcatcttgga ctgtattttt aggttggcta tgttcccagc 1620catatccctc cggggattca tgttgtgcag aaccaccagc acagtgtatc cggtgcactt 1680gggaaatttg tcatgtagct tagaaggaaa tgcgtggaag aacttggaga cgcccttgtg 1740acctccaaga ttttccatgc attcgtccat aatgatggca atgggcccac gggcggcggc 1800ctgggcgaag atatttctgg gatcactaac gtcatagttg tgttccagga tgagatcgtc 1860ataggccatt tttacaaagc gcgggcggag ggtgccagac tgcggtataa tggttccatc 1920cggcccaggg gcgtagttac cctcacagat ttaagggtgg gaaagaatat ataaggtggg 1980ggtcttatgt agttttgtat ctgttttgca gcagccgccg ccgccatgag caccaactcg 2040tttgatggaa gcattgtgag ctcatatttg acaacgcgca tgcccccatg ggccggggtg 2100cgtcagaatg tgatgggctc cagcattgat ggtcgccccg tcctgcccgc aaactctact 2160accttgacct acgagaccgt gtctggaacg ccgttggaga ctgcagcctc cgccgccgct 2220tcagccgctg cagccaccgc ccgcgggatt gtgactgact ttgctttcct gagcccgctt 2280gcaagcagtg cagcttcccg ttcatccgcc cgcgatgaca agttgacggc tcttttggca 2340caattggatt ctttgacccg ggaacttaat gtcgtttctc agcagctgtt ggatctgcgc 2400cagcaggttt ctgccctgaa ggcttcctcc cctcccaatg cggtttaaaa cataaataaa 2460aaaccagact ctgtttggat ttggatcaag caagtgtctt gctgtcttta tttaggggtt 2520ttgcgcgcgc ggtaggcccg ggaccagcgg tctcggtcgt tgagggtcct gtgtattttt 2580tccaggacgt ggtaaaggtg actctggatg ttcagataca tgggcataag cccgtctctg 2640gggtggaggt agcaccactg cagagcttca tgctgcgggg tggtgttgta gatgatccag 2700tcgtagcagg agcgctgggc gtggtgccta aaaatgtctt tcagtagcaa gctgattgcc 2760aggggcaggc ccttggtgta agtgtttaca aagcggttaa gctgggatgg gtgcatacgt 2820ggggatatga gatgcatctt ggactgtatt tttaggttgg ctatgttccc agccatatcc 2880ctccggggat tcatgttgtg cagaaccacc agcacagtgt atccggtgca cttgggaaat 2940ttgtcatgta gcttagaagg aaatgcgtgg aagaacttgg agacgccctt gtgacctcca 3000agattttcca tgcattcgtc cataatgatg gcaatgggcc cacgggcggc ggcctgggcg 3060aagatatttc tgggatcact aacgtcatag ttgtgttcca ggatgagatc gtcataggcc 3120atttttacaa agcgcgggcg gagggtgcca gactgcggta taatggttcc atccggccca 3180ggggcgtagt taccctcaca gatttgcatt tcccacgctt tgagttcaga tggggggatc 3240atgtctacct gcggggcgat gaagaaaacg gtttccgggg taggggagat cagctgggaa 3300gaaagcaggt tcctgagcag ctgcgactta ccgcagccgg tgggcccgta aatcacacct 3360attaccggct gcaactggta gttaagagag ctgcagctgc cgtcatccct gagcaggggg 3420gccacttcgt taagcatgtc cctgactcgc atgttttccc tgaccaaatc cgccagaagg 3480cgctcgccgc ccagcgatag cagttcttgc aaggaagcaa agtttttcaa cggtttgaga 3540ccgtccgccg taggcatgct tttgagcgtt tgaccaagca gttccaggcg gtcccacagc 3600tcggtcacct gctctacggc atctcgatcc agcatatctc ctcgtttcgc gggttggggc 3660ggctttcgct gtacggcagt agtcggtgct cgtccagacg ggccagggtc atgtctttcc 3720acgggcgcag ggtcctcgtc agcgtagtct gggtcacggt gaaggggtgc gctccgggct 3780gcgcgctggc cagggtgcgc ttgaggctgg tcctgctggt gctgaagcgc tgccggtctt 3840cgccctgcgc gtcggccagg tagcatttga ccatggtgtc atagtccagc ccctccgcgg 3900cgtggccctt ggcgcgcagc ttgcccttgg aggaggcgcc gcacgagggg cagtgcagac 3960ttttgagggc gtagagcttg ggcgcgagaa ataccgattc cggggagtag gcatccgcgc 4020cgcaggcccc gcagacggtc tcgcattcca cgagccaggt gagctctggc cgttcggggt 4080caaaaaccag gtttccccca tgctttttga tgcgtttctt acctctggtt tccatgagcc 4140ggtgtccacg ctcggtgacg aaaaggctgt ccgtgtcccc gtatacagac ttgagaggcc 4200tgtcctcgag cggtgttccg cggtcctcct cgtatagaaa ctcggaccac tctgagacaa 4260aggctcgcgt ccaggccagc acgaaggagg ctaagtggga ggggtagcgg tcgttgtcca 4320ctagggggtc cactcgctcc agggtgtgaa gacacatgtc gccctcttcg gcatcaagga 4380aggtgattgg tttgtaggtg taggccacgt gaccgggtgt tcctgaaggg gggctataaa 4440agggggtggg ggcgcgttcg tcctcactct cttccgcatc gctgtctgcg agggccagct 4500gttggggtga gtactccctc tgaaaagcgg gcatgacttc tgcgctaaga ttgtcagttt 4560ccaaaaacga ggaggatttg atattcacct ggcccgcggt gatgcctttg agggtggccg 4620catccatctg gtcagaaaag acaatctttt tgttgtcaag cttggtggca aacgacccgt 4680agagggcgtt ggacagcaac ttggcgatgg agcgcagggt ttggtttttg tcgcgatcgg 4740cgcgctcctt ggccgcgatg tttagctgca cgtattcgcg cgcaacgcac cgccattcgg 4800gaaagacggt ggtgcgctcg tcgggcacca ggtgcacgcg ccaaccgcgg ttgtgcaggg 4860tgacaaggtc aacgctggtg gctacctctc cgcgtaggcg ctcgttggtc cagcagaggc 4920ggccgccctt gcgcgagcag aatggcggta gggggtctag ctgcgtctcg tccggggggt 4980ctgcgtccac ggtaaagacc ccgggcagca ggcgcgcgtc gaagtagtct atcttgcatc 5040cttgcaagtc tagcgcctgc tgccatgcgc gggcggcaag cgcgcgctcg tatgggttga 5100gtgggggacc ccatggcatg gggtgggtga gcgcggaggc gtacatgccg caaatgtcgt 5160aaacgtagag gggctctctg agtattccaa gatatgtagg gtagcatctt ccaccgcgga 5220tgctggcgcg cacgtaatcg tatagttcgt gcgagggagc gaggaggtcg ggaccgaggt 5280tgctacgggc gggctgctct gctcggaaga ctatctgcct gaagatggca tgtgagttgg 5340atgatatggt tggacgctgg aagacgttga agctggcgtc tgtgagacct accgcgtcac 5400gcacgaagga ggcgtaggag tcgcgcagct tgttgaccag ctcggcggtg acctgcacgt 5460ctagggcgca gtagtccagg gtttccttga tgatgtcata cttatcctgt cccttttttt 5520tccacagctc gcggttgagg acaaactctt cgcggtcttt ccagtactct tggatcggaa 5580acccgtcggc ctccgaacgg taagagccta gcatgtagaa ctggttgacg gcctggtagg 5640cgcagcatcc cttttctacg ggtagcgcgt atgcctgcgc ggccttccgg agcgaggtgt 5700gggtgagcgc aaaggtgtcc ctgaccatga ctttgaggta ctggtatttg aagtcagtgt 5760cgtcgcatcc gccctgctcc cagagcaaaa agtccgtgcg ctttttggaa cgcggatttg 5820gcagggcgaa ggtgacatcg ttgaagagta tctttcccgc gcgaggcata aagttgcgtg 5880tgatgcggaa gggtcccggc acctcggaac ggttgttaat tacctgggcg gcgagcacga 5940tctcgtcaaa gccgttgatg ttgtggccca caatgtaaag ttccaagaag cgcgggatgc 6000ccttgatgga aggcaatttt ttaagttcct cgtaggtgag ctcttcaggg gagctgagcc 6060cgtgctctga aagggcccag tctgcaagat gagggttgga agcgacgaat gagctccaca 6120ggtcacgggc cattagcatt tgcaggtggt cgcgaaaggt cctaaactgg cgacctatgg 6180ccattttttc tggggtgatg cagtagaagg taagcgggtc ttgttcccag cggtcccatc 6240caaggttcgc ggctaggtct cgcgcggcag tcactagagg ctcatctccg ccgaacttca 6300tgaccagcat gaagggcacg agctgcttcc caaaggcccc catccaagta taggtctcta 6360catcgtaggt gacaaagaga cgctcggtgc gaggatgcga gccgatcggg aagaactgga 6420tctcccgcca ccaattggag gagtggctat tgatgtggtg aaagtagaag tccctgcgac 6480gggccgaaca ctcgtgctgg cttttgtaaa aacgtgcgca gtactggcag cggtgcacgg 6540gctgtacatc ctgcacgagg ttgacctgac gaccgcgcac aaggaagcag agtgggaatt 6600tgagcccctc gcctggcggg tttggctggt ggtcttctac ttcggctgct tgtccttgac 6660cgtctggctg ctcgagggga gttacggtgg atcggaccac cacgccgcgc gagcccaaag 6720tccagatgtc cgcgcgcggc ggtcggagct tgatgacaac atcgcgcaga tgggagctgt 6780ccatggtctg gagctcccgc ggcgtcaggt caggcgggag ctcctgcagg tttacctcgc 6840atagacgggt cagggcgcgg gctagatcca ggtgatacct aatttccagg ggctggttgg 6900tggcggcgtc gatggcttgc aagaggccgc atccccgcgg cgcgactacg gtaccgcgcg 6960gcgggcggtg ggccgcgggg gtgtccttgg atgatgcatc taaaagcggt gacgcgggcg 7020agcccccgga ggtagggggg gctccggacc cgccgggaga gggggcaggg gcacgtcggc 7080gccgcgcgcg ggcaggagct ggtgctgcgc gcgtaggttg ctggcgaacg cgacgacgcg 7140gcggttgatc tcctgaatct ggcgcctctg cgtgaagacg acgggcccgg tgagcttgaa 7200cctgaaagag agttcgacag aatcaatttc ggtgtcgttg acggcggcct ggcgcaaaat 7260ctcctgcacg tctcctgagt tgtcttgata ggcgatctcg gccatgaact gctcgatctc 7320ttcctcctgg agatctccgc gtccggctcg ctccacggtg gcggcgaggt cgttggaaat 7380gcgggccatg agctgcgaga aggcgttgag gcctccctcg ttccagacgc ggctgtagac 7440cacgccccct tcggcatcgc gggcgcgcat gaccacctgc gcgagattga gctccacgtg 7500ccgggcgaag acggcgtagt ttcgcaggcg ctgaaagagg tagttgaggg tggtggcggt 7560gtgttctgcc acgaagaagt acataaccca gcgtcgcaac gtggattcgt tgatatcccc 7620caaggcctca aggcgctcca tggcctcgta gaagtccacg gcgaagttga aaaactggga 7680gttgcgcgcc gacacggtta actcctcctc cagaagacgg atgagctcgg cgacagtgtc 7740gcgcacctcg cgctcaaagg ctacaggggc ctcttcttct tcttcaatct cctcttccat 7800aagggcctcc ccttcttctt cttctggcgg cggtggggga ggggggacac ggcggcgacg 7860acggcgcacc gggaggcggt cgacaaagcg ctcgatcatc tccccgcggc gacggcgcat 7920ggtctcggtg acggcgcggc cgttctcgcg ggggcgcagt tggaagacgc cgcccgtcat 7980gtcccggtta tgggttggcg gggggctgcc atgcggcagg gatacggcgc taacgatgca 8040tctcaacaat tgttgtgtag gtactccgcc gccgagggac ctgagcgagt ccgcatcgac 8100cggatcggaa aacctctcga gaaaggcgtc taaccagtca cagtcgcaag gtaggctgag 8160caccgtggcg ggcggcagcg ggcggcggtc ggggttgttt ctggcggagg tgctgctgat 8220gatgtaatta aagtaggcgg tcttgagacg gcggatggtc gacagaagca ccatgtcctt 8280gggtccggcc tgctgaatgc gcaggcggtc ggccatgccc caggcttcgt tttgacatcg 8340gcgcaggtct ttgtagtagt cttgcatgag cctttctacc ggcacttctt cttctccttc 8400ctcttgtcct gcatctcttg catctatcgc tgcggcggcg gcggagtttg gccgtaggtg 8460gcgccctctt cctcccatgc gtgtgacccc gaagcccctc atcggctgaa gcagggctag 8520gtcggcgaca acgcgctcgg ctaatatggc ctgctgcacc tgcgtgaggg tagactggaa 8580gtcatccatg tccacaaagc ggtggtatgc gcccgtgttg atggtgtaag tgcagttggc 8640cataacggac cagttaacgg tctggtgacc cggctgcgag agctcggtgt acctgagacg 8700cgagtaagcc ctcgagtcaa atacgtagtc gttgcaagtc cgcaccaggt actggtatcc 8760caccaaaaag tgcggcggcg gctggcggta gaggggccag cgtagggtgg ccggggctcc 8820gggggcgaga tcttccaaca taaggcgatg atatccgtag atgtacctgg acatccaggt 8880gatgccggcg gcggtggtgg aggcgcgcgg aaagtcgcgg acgcggttcc agatgttgcg 8940cagcggcaaa aagtgctcca tggtcgggac gctctggccg gtcaggcgcg cgcaatcgtt 9000gacgctctag accgtgcaaa aggagagcct gtaagcgggc actcttccgt ggtctggtgg 9060ataaattcgc aagggtatca tggcggacga ccggggttcg agccccgtat ccggccgtcc 9120gccgtgatcc atgcggttac cgcccgcgtg tcgaacccag gtgtgcgacg tcagacaacg 9180ggggagtgct ccttttggct tccttccagg cgcggcggct gctgcgctag cttttttggc 9240cactggccgc gcgcagcgta agcggttagg ctggaaagcg aaagcattaa gtggctcgct 9300ccctgtagcc ggagggttat tttccaaggg ttgagtcgcg ggacccccgg ttcgagtctc 9360ggaccggccg gactgcggcg aacgggggtt tgcctccccg tcatgcaaga ccccgcttgc 9420aaattcctcc ggaaacaggg acgagcccct tttttgcttt tcccagatgc atccggtgct 9480gcggcagatg cgcccccctc ctcagcagcg gcaagagcaa gagcagcggc agacatgcag 9540ggcaccctcc cctcctccta ccgcgtcagg aggggcgaca tccgcggttg acgcggcagc 9600agatggtgat tacgaacccc cgcggcgccg ggcccggcac tacctggact tggaggaggg 9660cgagggcctg gcgcggctag gagcgccctc tcctgagcgg cacccaaggg tgcagctgaa 9720gcgtgatacg cgtgaggcgt acgtgccgcg gcagaacctg tttcgcgacc gcgagggaga 9780ggagcccgag gagatgcggg atcgaaagtt ccacgcaggg cgcgagctgc ggcatggcct 9840gaatcgcgag cggttgctgc gcgaggagga ctttgagccc gacgcgcgaa ccgggattag 9900tcccgcgcgc gcacacgtgg cggccgccga cctggtaacc gcatacgagc agacggtgaa 9960ccaggagatt aactttcaaa aaagctttaa caaccacgtg cgtacgcttg tggcgcgcga 10020ggaggtggct ataggactga tgcatctgtg ggactttgta agcgcgctgg agcaaaaccc 10080aaatagcaag ccgctcatgg cgcagctgtt ccttatagtg cagcacagca gggacaacga 10140ggcattcagg gatgcgctgc taaacatagt agagcccgag ggccgctggc tgctcgattt 10200gataaacatc ctgcagagca tagtggtgca ggagcgcagc ttgagcctgg ctgacaaggt 10260ggccgccatc aactattcca tgcttagcct gggcaagttt tacgcccgca agatatacca 10320taccccttac gttcccatag acaaggaggt aaagatcgag gggttctaca tgcgcatggc 10380gctgaaggtg cttaccttga gcgacgacct gggcgtttat cgcaacgagc gcatccacaa 10440ggccgtgagc gtgagccggc ggcgcgagct cagcgaccgc gagctgatgc acagcctgca 10500aagggccctg gctggcacgg gcagcggcga tagagaggcc gagtcctact ttgacgcggg 10560cgctgacctg cgctgggccc caagccgacg cgccctggag gcagctgggg ccggacctgg 10620gctggcggtg gcacccgcgc gcgctggcaa cgtcggcggc gtggaggaat atgacgagga 10680cgatgagtac gagccagagg acggcgagta ctaagcggtg atgtttctga tcagatgatg 10740caagacgcaa cggacccggc ggtgcgggcg gcgctgcaga gccagccgtc cggccttaac 10800tccacggacg actggcgcca ggtcatggac cgcatcatgt cgctgactgc gcgcaatcct 10860gacgcgttcc ggcagcagcc gcaggccaac cggctctccg caattctgga agcggtggtc 10920ccggcgcgcg caaaccccac gcacgagaag gtgctggcga tcgtaaacgc gctggccgaa 10980aacagggcca tccggcccga cgaggccggc ctggtctacg acgcgctgct tcagcgcgtg 11040gctcgttaca acagcggcaa cgtgcagacc aacctggacc ggctggtggg ggatgtgcgc 11100gaggccgtgg cgcagcgtga gcgcgcgcag cagcagggca acctgggctc catggttgca 11160ctaaacgcct tcctgagtac acagcccgcc aacgtgccgc ggggacagga ggactacacc 11220aactttgtga gcgcactgcg gctaatggtg actgagacac cgcaaagtga ggtgtaccag 11280tctgggccag actatttttt ccagaccagt agacaaggcc tgcagaccgt aaacctgagc 11340caggctttca aaaacttgca ggggctgtgg ggggtgcggg ctcccacagg cgaccgcgcg 11400accgtgtcta gcttgctgac gcccaactcg cgcctgttgc tgctgctaat agcgcccttc 11460acggacagtg gcagcgtgtc ccgggacaca tacctaggtc acttgctgac actgtaccgc 11520gaggccatag gtcaggcgca tgtggacgag catactttcc aggagattac aagtgtcagc 11580cgcgcgctgg ggcaggagga cacgggcagc ctggaggcaa ccctaaacta cctgctgacc 11640aaccggcggc agaagatccc ctcgttgcac agtttaaaca gcgaggagga gcgcattttg 11700cgctacgtgc agcagagcgt gagccttaac ctgatgcgcg acggggtaac gcccagcgtg 11760gcgctggaca tgaccgcgcg caacatggaa ccgggcatgt atgcctcaaa ccggccgttt 11820atcaaccgcc taatggacta cttgcatcgc gcggccgccg tgaaccccga gtatttcacc 11880aatgccatct tgaacccgca ctggctaccg ccccctggtt tctacaccgg gggattcgag 11940gtgcccgagg gtaacgatgg attcctctgg gacgacatag acgacagcgt gttttccccg 12000caaccgcaga ccctgctaga gttgcaacag cgcgagcagg cagaggcggc gctgcgaaag 12060gaaagcttcc gcaggccaag cagcttgtcc gatctaggcg ctgcggcccc gcggtcagat 12120gctagtagcc catttccaag cttgataggg tctcttacca gcactcgcac cacccgcccg 12180cgcctgctgg gcgaggagga gtacctaaac aactcgctgc tgcagccgca gcgcgaaaaa 12240aacctgcctc cggcatttcc caacaacggg atagagagcc tagtggacaa gatgagtaga 12300tggaagacgt acgcgcagga gcacagggac gtgccaggcc cgcgcccgcc cacccgtcgt 12360caaaggcacg accgtcagcg gggtctggtg tgggaggacg atgactcggc agacgacagc 12420agcgtcctgg atttgggagg gagtggcaac ccgtttgcgc accttcgccc caggctgggg 12480agaatgtttt aaaaaaaaaa aaagcatgat gcaaaataaa aaactcacca aggccatggc 12540accgagcgtt ggttttcttg tattcccctt agtatgcggc gcgcggcgat gtatgaggaa 12600ggtcctcctc cctcctacga gagtgtggtg agcgcggcgc cagtggcggc ggcgctgggt 12660tctcccttcg atgctcccct ggacccgccg tttgtgcctc cgcggtacct gcggcctacc 12720ggggggagaa acagcatccg ttactctgag ttggcacccc tattcgacac cacccgtgtg 12780tacctggtgg acaacaagtc aacggatgtg gcatccctga actaccagaa cgaccacagc 12840aactttctga ccacggtcat tcaaaacaat gactacagcc cgggggaggc aagcacacag 12900accatcaatc ttgacgaccg gtcgcactgg ggcggcgacc tgaaaaccat cctgcatacc 12960aacatgccaa atgtgaacga gttcatgttt accaataagt ttaaggcgcg ggtgatggtg 13020tcgcgcttgc ctactaagga caatcaggtg gagctgaaat acgagtgggt ggagttcacg 13080ctgcccgagg gcaactactc cgagaccatg accatagacc ttatgaacaa cgcgatcgtg 13140gagcactact tgaaagtggg cagacagaac ggggttctgg aaagcgacat cggggtaaag 13200tttgacaccc gcaacttcag actggggttt gaccccgtca ctggtcttgt catgcctggg 13260gtatatacaa acgaagcctt ccatccagac atcattttgc tgccaggatg cggggtggac 13320ttcacccaca gccgcctgag caacttgttg ggcatccgca agcggcaacc cttccaggag 13380ggctttagga tcacctacga tgatctggag ggtggtaaca ttcccgcact gttggatgtg 13440gacgcctacc aggcgagctt gaaagatgac accgaacagg gcgggggtgg cgcaggcggc 13500agcaacagca gtggcagcgg cgcggaagag aactccaacg cggcagccgc ggcaatgcag 13560ccggtggagg acatgaacga tcatgccatt cgcggcgaca cctttgccac acgggctgag 13620gagaagcgcg ctgaggccga agcagcggcc gaagctgccg cccccgctgc gcaacccgag 13680gtcgagaagc ctcagaagaa accggtgatc aaacccctga cagaggacag caagaaacgc 13740agttacaacc taataagcaa tgacagcacc ttcacccagt accgcagctg gtaccttgca 13800tacaactacg gcgaccctca gaccggaatc cgctcatgga ccctgctttg cactcctgac 13860gtaacctgcg gctcggagca ggtctactgg tcgttgccag acatgatgca agaccccgtg 13920accttccgct ccacgcgcca gatcagcaac tttccggtgg tgggcgccga gctgttgccc 13980gtgcactcca agagcttcta caacgaccag gccgtctact cccaactcat ccgccagttt 14040acctctctga cccacgtgtt caatcgcttt cccgagaacc agattttggc gcgcccgcca 14100gcccccacca tcaccaccgt cagtgaaaac gttcctgctc tcacagatca cgggacgcta 14160ccgctgcgca acagcatcgg aggagtccag cgagtgacca ttactgacgc cagacgccgc 14220acctgcccct acgtttacaa ggccctgggc atagtctcgc cgcgcgtcct atcgagccgc 14280actttttgag caagcatgtc catccttata tcgcccagca ataacacagg ctggggcctg 14340cgcttcccaa gcaagatgtt tggcggggcc aagaagcgct ccgaccaaca cccagtgcgc 14400gtgcgcgggc actaccgcgc gccctggggc gcgcacaaac gcggccgcac tgggcgcacc 14460accgtcgatg acgccatcga cgcggtggtg gaggaggcgc gcaactacac gcccacgccg 14520ccaccagtgt ccacagtgga cgcggccatt cagaccgtgg tgcgcggagc ccggcgctat 14580gctaaaatga agagacggcg gaggcgcgta gcacgtcgcc accgccgccg acccggcact 14640gccgcccaac gcgcggcggc ggccctgctt aaccgcgcac gtcgcaccgg ccgacgggcg 14700gccatgcggg ccgctcgaag gctggccgcg ggtattgtca ctgtgccccc caggtccagg 14760cgacgagcgg ccgccgcagc agccgcggcc attagtgcta tgactcaggg tcgcaggggc 14820aacgtgtatt gggtgcgcga ctcggttagc ggcctgcgcg tgcccgtgcg cacccgcccc 14880ccgcgcaact agattgcaag aaaaaactac ttagactcgt actgttgtat gtatccagcg 14940gcggcggcgc gcaacgaagc tatgtccaag cgcaaaatca aagaagagat gctccaggtc

15000atcgcgccgg agatctatgg ccccccgaag aaggaagagc aggattacaa gccccgaaag 15060ctaaagcggg tcaaaaagaa aaagaaagat gatgatgatg aacttgacga cgaggtggaa 15120ctgctgcacg ctaccgcgcc caggcgacgg gtacagtgga aaggtcgacg cgtaaaacgt 15180gttttgcgac ccggcaccac cgtagtcttt acgcccggtg agcgctccac ccgcacctac 15240aagcgcgtgt atgatgaggt gtacggcgac gaggacctgc ttgagcaggc caacgagcgc 15300ctcggggagt ttgcctacgg aaagcggcat aaggacatgc tggcgttgcc gctggacgag 15360ggcaacccaa cacctagcct aaagcccgta acactgcagc aggtgctgcc cgcgcttgca 15420ccgtccgaag aaaagcgcgg cctaaagcgc gagtctggtg acttggcacc caccgtgcag 15480ctgatggtac ccaagcgcca gcgactggaa gatgtcttgg aaaaaatgac cgtggaacct 15540gggctggagc ccgaggtccg cgtgcggcca atcaagcagg tggcgccggg actgggcgtg 15600cagaccgtgg acgttcagat acccactacc agtagcacca gtattgccac cgccacagag 15660ggcatggaga cacaaacgtc cccggttgcc tcagcggtgg cggatgccgc ggtgcaggcg 15720gtcgctgcgg ccgcgtccaa gacctctacg gaggtgcaaa cggacccgtg gatgtttcgc 15780gtttcagccc cccggcgccc gcgccgttcg aggaagtacg gcgccgccag cgcgctactg 15840cccgaatatg ccctacatcc ttccattgcg cctacccccg gctatcgtgg ctacacctac 15900cgccccagaa gacgagcaac tacccgacgc cgaaccacca ctggaacccg ccgccgccgt 15960cgccgtcgcc agcccgtgct ggccccgatt tccgtgcgca gggtggctcg cgaaggaggc 16020aggaccctgg tgctgccaac agcgcgctac caccccagca tcgtttaaaa gccggtcttt 16080gtggttcttg cagatatggc cctcacctgc cgcctccgtt tcccggtgcc gggattccga 16140ggaagaatgc accgtaggag gggcatggcc ggccacggcc tgacgggcgg catgcgtcgt 16200gcgcaccacc ggcggcggcg cgcgtcgcac cgtcgcatgc gcggcggtat cctgcccctc 16260cttattccac tgatcgccgc ggcgattggc gccgtgcccg gaattgcatc cgtggccttg 16320caggcgcaga gacactgatt aaaaacaagt tgcatgtgga aaaatcaaaa taaaaagtct 16380ggactctcac gctcgcttgg tcctgtaact attttgtaga atggaagaca tcaactttgc 16440gtctctggcc ccgcgacacg gctcgcgccc gttcatggga aactggcaag atatcggcac 16500cagcaatatg agcggtggcg ccttcagctg gggctcgctg tggagcggca ttaaaaattt 16560cggttccacc gttaagaact atggcagcaa ggcctggaac agcagcacag gccagatgct 16620gagggataag ttgaaagagc aaaatttcca acaaaaggtg gtagatggcc tggcctctgg 16680cattagcggg gtggtggacc tggccaacca ggcagtgcaa aataagatta acagtaagct 16740tgatccccgc cctcccgtag aggagcctcc accggccgtg gagacagtgt ctccagaggg 16800gcgtggcgaa aagcgtccgc gccccgacag ggaagaaact ctggtgacgc aaatagacga 16860gcctccctcg tacgaggagg cactaaagca aggcctgccc accacccgtc ccatcgcgcc 16920catggctacc ggagtgctgg gccagcacac acccgtaacg ctggacctgc ctccccccgc 16980cgacacccag cagaaacctg tgctgccagg cccgaccgcc gttgttgtaa cccgtcctag 17040ccgcgcgtcc ctgcgccgcg ccgccagcgg tccgcgatcg ttgcggcccg tagccagtgg 17100caactggcaa agcacactga acagcatcgt gggtctgggg gtgcaatccc tgaagcgccg 17160acgatgcttc tgatagctaa cgtgtcgtat gtgtgtcatg tatgcgtcca tgtcgccgcc 17220agaggagctg ctgagccgcc gcgcgcccgc tttccaagat ggctacccct tcgatgatgc 17280cgcagtggtc ttacatgcac atctcgggcc aggacgcctc ggagtacctg agccccgggc 17340tggtgcagtt tgcccgcgcc accgagacgt acttcagcct gaataacaag tttagaaacc 17400ccacggtggc gcctacgcac gacgtgacca cagaccggtc ccagcgtttg acgctgcggt 17460tcatccctgt ggaccgtgag gatactgcgt actcgtacaa ggcgcggttc accctagctg 17520tgggtgataa ccgtgtgctg gacatggctt ccacgtactt tgacatccgc ggcgtgctgg 17580acaggggccc tacttttaag ccctactctg gcactgccta caacgccctg gctcccaagg 17640gtgccccaaa tccttgcgaa tgggatgaag ctgctactgc tcttgaaata aacctagaag 17700aagaggacga tgacaacgaa gacgaagtag acgagcaagc tgagcagcaa aaaactcacg 17760tatttgggca ggcgccttat tctggtataa atattacaaa ggagggtatt caaataggtg 17820tcgaaggtca aacacctaaa tatgccgata aaacatttca acctgaacct caaataggag 17880aatctcagtg gtacgaaaca gaaattaatc atgcagctgg gagagtccta aaaaagacta 17940ccccaatgaa accatgttac ggttcatatg caaaacccac aaatgaaaat ggagggcaag 18000gcattcttgt aaagcaacaa aatggaaagc tagaaagtca agtggaaatg caatttttct 18060caactactga ggcagccgca ggcaatggtg ataacttgac tcctaaagtg gtattgtaca 18120gtgaagatgt agatatagaa accccagaca ctcatatttc ttacatgccc actattaagg 18180aaggtaactc acgagaacta atgggccaac aatctatgcc caacaggcct aattacattg 18240cttttaggga caattttatt ggtctaatgt attacaacag cacgggtaat atgggtgttc 18300tggcgggcca agcatcgcag ttgaatgctg ttgtagattt gcaagacaga aacacagagc 18360tttcatacca gcttttgctt gattccattg gtgatagaac caggtacttt tctatgtgga 18420atcaggctgt tgacagctat gatccagatg ttagaattat tgaaaatcat ggaactgaag 18480atgaacttcc aaattactgc tttccactgg gaggtgtgat taatacagag actcttacca 18540aggtaaaacc taaaacaggt caggaaaatg gatgggaaaa agatgctaca gaattttcag 18600ataaaaatga aataagagtt ggaaataatt ttgccatgga aatcaatcta aatgccaacc 18660tgtggagaaa tttcctgtac tccaacatag cgctgtattt gcccgacaag ctaaagtaca 18720gtccttccaa cgtaaaaatt tctgataacc caaacaccta cgactacatg aacaagcgag 18780tggtggctcc cgggctagtg gactgctaca ttaaccttgg agcacgctgg tcccttgact 18840atatggacaa cgtcaaccca tttaaccacc accgcaatgc tggcctgcgc taccgctcaa 18900tgttgctggg caatggtcgc tatgtgccct tccacatcca ggtgcctcag aagttctttg 18960ccattaaaaa cctccttctc ctgccgggct catacaccta cgagtggaac ttcaggaagg 19020atgttaacat ggttctgcag agctccctag gaaatgacct aagggttgac ggagccagca 19080ttaagtttga tagcatttgc ctttacgcca ccttcttccc catggcccac aacaccgcct 19140ccacgcttga ggccatgctt agaaacgaca ccaacgacca gtcctttaac gactatctct 19200ccgccgccaa catgctctac cctatacccg ccaacgctac caacgtgccc atatccatcc 19260cctcccgcaa ctgggcggct ttccgcggct gggccttcac gcgccttaag actaaggaaa 19320ccccatcact gggctcgggc tacgaccctt attacaccta ctctggctct ataccctacc 19380tagatggaac cttttacctc aaccacacct ttaagaaggt ggccattacc tttgactctt 19440ctgtcagctg gcctggcaat gaccgcctgc ttacccccaa cgagtttgaa attaagcgct 19500cagttgacgg ggagggttac aacgttgccc agtgtaacat gaccaaagac tggttcctgg 19560tacaaatgct agctaactat aacattggct accagggctt ctatatccca gagagctaca 19620aggaccgcat gtactccttc tttagaaact tccagcccat gagccgtcag gtggtggatg 19680atactaaata caaggactac caacaggtgg gcatcctaca ccaacacaac aactctggat 19740ttgttggcta ccttgccccc accatgcgcg aaggacaggc ctaccctgct aacttcccct 19800atccgcttat aggcaagacc gcagttgaca gcattaccca gaaaaagttt ctttgcgatc 19860gcaccctttg gcgcatccca ttctccagta actttatgtc catgggcgca ctcacagacc 19920tgggccaaaa ccttctctac gccaactccg cccacgcgct agacatgact tttgaggtgg 19980atcccatgga cgagcccacc cttctttatg ttttgtttga agtctttgac gtggtccgtg 20040tgcaccagcc gcaccgcggc gtcatcgaaa ccgtgtacct gcgcacgccc ttctcggccg 20100gcaacgccac aacataaaga agcaagcaac atcaacaaca gctgccgcca tgggctccag 20160tgagcaggaa ctgaaagcca ttgtcaaaga tcttggttgt gggccatatt ttttgggcac 20220ctatgacaag cgctttccag gctttgtttc tccacacaag ctcgcctgcg ccatagtcaa 20280tacggccggt cgcgagactg ggggcgtaca ctggatggcc tttgcctgga acccgcactc 20340aaaaacatgc tacctctttg agccctttgg cttttctgac cagcgactca agcaggttta 20400ccagtttgag tacgagtcac tcctgcgccg tagcgccatt gcttcttccc ccgaccgctg 20460tataacgctg gaaaagtcca cccaaagcgt acaggggccc aactcggccg cctgtggact 20520attctgctgc atgtttctcc acgcctttgc caactggccc caaactccca tggatcacaa 20580ccccaccatg aaccttatta ccggggtacc caactccatg ctcaacagtc cccaggtaca 20640gcccaccctg cgtcgcaacc aggaacagct ctacagcttc ctggagcgcc actcgcccta 20700cttccgcagc cacagtgcgc agattaggag cgccacttct ttttgtcact tgaaaaacat 20760gtaaaaataa tgtactagag acactttcaa taaaggcaaa tgcttttatt tgtacactct 20820cgggtgatta tttaccccca cccttgccgt ctgcgccgtt taaaaatcaa aggggttctg 20880ccgcgcatcg ctatgcgcca ctggcaggga cacgttgcga tactggtgtt tagtgctcca 20940cttaaactca ggcacaacca tccgcggcag ctcggtgaag ttttcactcc acaggctgcg 21000caccatcacc aacgcgttta gcaggtcggg cgccgatatc ttgaagtcgc agttggggcc 21060tccgccctgc gcgcgcgagt tgcgatacac agggttgcag cactggaaca ctatcagcgc 21120cgggtggtgc acgctggcca gcacgctctt gtcggagatc agatccgcgt ccaggtcctc 21180cgcgttgctc agggcgaacg gagtcaactt tggtagctgc cttcccaaaa agggcgcgtg 21240cccaggcttt gagttgcact cgcaccgtag tggcatcaaa aggtgaccgt gcccggtctg 21300ggcgttagga tacagcgcct gcataaaagc cttgatctgc ttaaaagcca cctgagcctt 21360tgcgccttca gagaagaaca tgccgcaaga cttgccggaa aactgattgg ccggacaggc 21420cgcgtcgtgc acgcagcacc ttgcgtcggt gttggagatc tgcaccacat ttcggcccca 21480ccggttcttc acgatcttgg ccttgctaga ctgctccttc agcgcgcgct gcccgttttc 21540gctcgtcaca tccatttcaa tcacgtgctc cttatttatc ataatgcttc cgtgtagaca 21600cttaagctcg ccttcgatct cagcgcagcg gtgcagccac aacgcgcagc ccgtgggctc 21660gtgatgcttg taggtcacct ctgcaaacga ctgcaggtac gcctgcagga atcgccccat 21720catcgtcaca aaggtcttgt tgctggtgaa ggtcagctgc aacccgcggt gctcctcgtt 21780cagccaggtc ttgcatacgg ccgccagagc ttccacttgg tcaggcagta gtttgaagtt 21840cgcctttaga tcgttatcca cgtggtactt gtccatcagc gcgcgcgcag cctccatgcc 21900cttctcccac gcagacacga tcggcacact cagcgggttc atcaccgtaa tttcactttc 21960cgcttcgctg ggctcttcct cttcctcttg cgtccgcata ccacgcgcca ctgggtcgtc 22020ttcattcagc cgccgcactg tgcgcttacc tcctttgcca tgcttgatta gcaccggtgg 22080gttgctgaaa cccaccattt gtagcgccac atcttctctt tcttcctcgc tgtccacgat 22140tacctctggt gatggcgggc gctcgggctt gggagaaggg cgcttctttt tcttcttggg 22200cgcaatggcc aaatccgccg ccgaggtcga tggccgcggg ctgggtgtgc gcggcaccag 22260cgcgtcttgt gatgagtctt cctcgtcctc ggactcgata cgccgcctca tccgcttttt 22320tgggggcgcc cggggaggcg gcggcgacgg ggacggggac gacacgtcct ccatggttgg 22380gggacgtcgc gccgcaccgc gtccgcgctc gggggtggtt tcgcgctgct cctcttcccg 22440actggccatt tccttctcct ataggcagaa aaagatcatg gagtcagtcg agaagaagga 22500cagcctaacc gccccctctg agttcgccac caccgcctcc accgatgccg ccaacgcgcc 22560taccaccttc cccgtcgagg cacccccgct tgaggaggag gaagtgatta tcgagcagga 22620cccaggtttt gtaagcgaag acgacgagga ccgctcagta ccaacagagg ataaaaagca 22680agaccaggac aacgcagagg caaacgagga acaagtcggg cggggggacg aaaggcatgg 22740cgactaccta gatgtgggag acgacgtgct gttgaagcat ctgcagcgcc agtgcgccat 22800tatctgcgac gcgttgcaag agcgcagcga tgtgcccctc gccatagcgg atgtcagcct 22860tgcctacgaa cgccacctat tctcaccgcg cgtacccccc aaacgccaag aaaacggcac 22920atgcgagccc aacccgcgcc tcaacttcta ccccgtattt gccgtgccag aggtgcttgc 22980cacctatcac atctttttcc aaaactgcaa gataccccta tcctgccgtg ccaaccgcag 23040ccgagcggac aagcagctgg ccttgcggca gggcgctgtc atacctgata tcgcctcgct 23100caacgaagtg ccaaaaatct ttgagggtct tggacgcgac gagaagcgcg cggcaaacgc 23160tctgcaacag gaaaacagcg aaaatgaaag tcactctgga gtgttggtgg aactcgaggg 23220tgacaacgcg cgcctagccg tactaaaacg cagcatcgag gtcacccact ttgcctaccc 23280ggcacttaac ctacccccca aggtcatgag cacagtcatg agtgagctga tcgtgcgccg 23340tgcgcagccc ctggagaggg atgcaaattt gcaagaacaa acagaggagg gcctacccgc 23400agttggcgac gagcagctag cgcgctggct tcaaacgcgc gagcctgccg acttggagga 23460gcgacgcaaa ctaatgatgg ccgcagtgct cgttaccgtg gagcttgagt gcatgcagcg 23520gttctttgct gacccggaga tgcagcgcaa gctagaggaa acattgcact acacctttcg 23580acagggctac gtacgccagg cctgcaagat ctccaacgtg gagctctgca acctggtctc 23640ctaccttgga attttgcacg aaaaccgcct tgggcaaaac gtgcttcatt ccacgctcaa 23700gggcgaggcg cgccgcgact acgtccgcga ctgcgtttac ttatttctat gctacacctg 23760gcagacggcc atgggcgttt ggcagcagtg cttggaggag tgcaacctca aggagctgca 23820gaaactgcta aagcaaaact tgaaggacct atggacggcc ttcaacgagc gctccgtggc 23880cgcgcacctg gcggacatca ttttccccga acgcctgctt aaaaccctgc aacagggtct 23940gccagacttc accagtcaaa gcatgttgca gaactttagg aactttatcc tagagcgctc 24000aggaatcttg cccgccacct gctgtgcact tcctagcgac tttgtgccca ttaagtaccg 24060cgaatgccct ccgccgcttt ggggccactg ctaccttctg cagctagcca actaccttgc 24120ctaccactct gacataatgg aagacgtgag cggtgacggt ctactggagt gtcactgtcg 24180ctgcaaccta tgcaccccgc accgctccct ggtttgcaat tcgcagctgc ttaacgaaag 24240tcaaattatc ggtacctttg agctgcaggg tccctcgcct gacgaaaagt ccgcggctcc 24300ggggttgaaa ctcactccgg ggctgtggac gtcggcttac cttcgcaaat ttgtacctga 24360ggactaccac gcccacgaga ttaggttcta cgaagaccaa tcccgcccgc ctaatgcgga 24420gcttaccgcc tgcgtcatta cccagggcca cattcttggc caattgcaag ccatcaacaa 24480agcccgccaa gagtttctgc tacgaaaggg acggggggtt tacttggacc cccagtccgg 24540cgaggagctc aacccaatcc ccccgccgcc gcagccctat cagcagcagc cgcgggccct 24600tgcttcccag gatggcaccc aaaaagaagc tgcagctgcc gccgccaccc acggacgagg 24660aggaatactg ggacagtcag gcagaggagg ttttggacga ggaggaggag gacatgatgg 24720aagactggga gagcctagac gaggaagctt ccgaggtcga agaggtgtca gacgaaacac 24780cgtcaccctc ggtcgcattc ccctcgccgg cgccccagaa atcggcaacc ggttccagca 24840tggctacaac ctccgctcct caggcgccgc cggcactgcc cgttcgccga cccaaccgta 24900gatgggacac cactggaacc agggccggta agtccaagca gccgccgccg ttagcccaag 24960agcaacaaca gcgccaaggc taccgctcat ggcgcgggca caagaacgcc atagttgctt 25020gcttgcaaga ctgtgggggc aacatctcct tcgcccgccg ctttcttctc taccatcacg 25080gcgtggcctt cccccgtaac atcctgcatt actaccgtca tctctacagc ccatactgca 25140ccggcggcag cggcagcaac agcagcggcc acacagaagc aaaggcgacc ggatagcaag 25200actctgacaa agcccaagaa atccacagcg gcggcagcag caggaggagg agcgctgcgt 25260ctggcgccca acgaacccgt atcgacccgc gagcttagaa acaggatttt tcccactctg 25320tatgctatat ttcaacagag caggggccaa gaacaagagc tgaaaataaa aaacaggtct 25380ctgcgatccc tcacccgcag ctgcctgtat cacaaaagcg aagatcagct tcggcgcacg 25440ctggaagacg cggaggctct cttcagtaaa tactgcgcgc tgactcttaa ggactagttt 25500cgcgcccttt ctcaaattta agcgcgaaaa ctacgtcatc tccagcggcc acacccggcg 25560ccagcacctg ttgtcagcgc cattatgagc aaggaaattc ccacgcccta catgtggagt 25620taccagccac aaatgggact tgcggctgga gctgcccaag actactcaac ccgaataaac 25680tacatgagcg cgggacccca catgatatcc cgggtcaacg gaatacgcgc ccaccgaaac 25740cgaattctcc tggaacaggc ggctattacc accacacctc gtaataacct taatccccgt 25800agttggcccg ctgccctggt gtaccaggaa agtcccgctc ccaccactgt ggtacttccc 25860agagacgccc aggccgaagt tcagatgact aactcagggg cgcagcttgc gggcggcttt 25920cgtcacaggg tgcggtcgcc cgggcagggt ataactcacc tgacaatcag agggcgaggt 25980attcagctca acgacgagtc ggtgagctcc tcgcttggtc tccgtccgga cgggacattt 26040cagatcggcg gcgccggccg ctcttcattc acgcctcgtc aggcaatcct aactctgcag 26100acctcgtcct ctgagccgcg ctctggaggc attggaactc tgcaatttat tgaggagttt 26160gtgccatcgg tctactttaa ccccttctcg ggacctcccg gccactatcc ggatcaattt 26220attcctaact ttgacgcggt aaaggactcg gcggacggct acgactgaat gttaagtgga 26280gaggcagagc aactgcgcct gaaacacctg gtccactgtc gccgccacaa gtgctttgcc 26340cgcgactccg gtgagttttg ctactttgaa ttgcccgagg atcatatcga gggcccggcg 26400cacggcgtcc ggcttaccgc ccagggagag cttgcccgta gcctgattcg ggagtttacc 26460cagcgccccc tgctagttga gcgggacagg ggaccctgtg ttctcactgt gatttgcaac 26520tgtcctaacc ctggattaca tcaagatctt tgttgccatc tctgtgctga gtataataaa 26580tacagaaatt aaaatatact ggggctccta tcgccatcct gtaaacgcca ccgtcttcac 26640ccgcccaagc aaaccaaggc gaaccttacc tggtactttt aacatctctc cctctgtgat 26700ttacaacagt ttcaacccag acggagtgag tctacgagag aacctctccg agctcagcta 26760ctccatcaga aaaaacacca ccctccttac ctgccgggaa cgtacgagtg cgtcaccggc 26820cgctgcacca cacctaccgc ctgaccgtaa accagacttt ttccggacag acctcaataa 26880ctctgtttac cagaacagga ggtgagctta gaaaaccctt agggtattag gccaaaggcg 26940cagctactgt ggggtttatg aacaattcaa gcaactctac gggctattct aattcaggtt 27000tctctagaat cggggttggg gttattctct gtcttgtgat tctctttatt cttatactaa 27060cgcttctctg cctaaggctc gccgcctgct gtgtgcacat ttgcatttat tgtcagcttt 27120ttaaacgctg gggtcgccac ccaagatgat taggtacata atcctaggtt tactcaccct 27180tgcgtcagcc cacggtacca cccaaaaggt ggattttaag gagccagcct gtaatgttac 27240attcgcagct gaagctaatg agtgcaccac tcttataaaa tgcaccacag aacatgaaaa 27300gctgcttatt cgccacaaaa acaaaattgg caagtatgct gtttatgcta tttggcagcc 27360aggtgacact acagagtata atgttacagt tttccagggt aaaagtcata aaacttttat 27420gtatactttt ccattttatg aaatgtgcga cattaccatg tacatgagca aacagtataa 27480gttgtggccc ccacaaaatt gtgtggaaaa cactggcact ttctgctgca ctgctatgct 27540aattacagtg ctcgctttgg tctgtaccct actctatatt aaatacaaaa gcagacgcag 27600ctttattgag gaaaagaaaa tgccttaatt tactaagtta caaagctaat gtcaccacta 27660actgctttac tcgctgcttg caaaacaaat tcaaaaagtt agcattataa ttagaatagg 27720atttaaaccc cccggtcatt tcctgctcaa taccattccc ctgaacaatt gactctatgt 27780gggatatgct ccagcgctac aaccttgaag tcaggcttcc tggatgtcag catctgactt 27840tggccagcac ctgtcccgcg gatttgttcc agtccaacta cagcgaccca ccctaacaga 27900gatgaccaac acaaccaacg cggccgccgc taccggactt acatctacca caaatacacc 27960ccaagtttct gcctttgtca ataactggga taacttgggc atgtggtggt tctccatagc 28020gcttatgttt gtatgcctta ttattatgtg gctcatctgc tgcctaaagc gcaaacgcgc 28080ccgaccaccc atctatagtc ccatcattgt gctacaccca aacaatgatg gaatccatag 28140attggacgga ctgaaacaca tgttcttttc tcttacagta tgattaaatg agacatgatt 28200cctcgagttt ttatattact gacccttgtt gcgctttttt tgtgcgtgct ccacattggc 28260tgcggtttct cacatcgaag tagactgcat tccagccttc acagtctatt tgctttacgg 28320atttgtcacc ctcacgctca tctgcagcct catcactgtg gtcatcgcct ttatccagtg 28380cattgactgg gtctgtgtgc gctttgcata tctcagacac catccccagt acagggacag 28440gactatagct gagcttctta gaattcttta attatgaaat ttactgtgac ttttctgctg 28500attatttgca ccctatctgc gttttgttcc ccgacctcca agcctcaaag acatatatca 28560tgcagattca ctcgtatatg gaatattcca agttgctaca atgaaaaaag cgatctttcc 28620gaagcctggt tatatgcaat catctctgtt atggtgttct gcagtaccat cttagcccta 28680gctatatatc cctaccttga cattggctgg aacgcaatag atgccatgaa ccacccaact 28740ttccccgcgc ccgctatgct tccactgcaa caagttgttg ccggcggctt tgtcccagcc 28800aatcagcctc gcccaccttc tcccaccccc actgaaatca gctactttaa tctaacagga 28860ggagatgact gacaccctag atctagaaat ggacggaatt attacagagc agcgcctgct 28920agaaagacgc agggcagcgg ccgagcaaca gcgcatgaat caagagctcc aagacatggt 28980taacttgcac cagtgcaaaa ggggtatctt ttgtctggta aagcaggcca aagtcaccta 29040cgacagtaat accaccggac accgccttag ctacaagttg ccaaccaagc gtcagaaatt 29100ggtggtcatg gtgggagaaa agcccattac cataactcag cactcggtag aaaccgaagg 29160ctgcattcac tcaccttgtc aaggacctga ggatctctgc acccttatta agaccctgtg 29220cggtctcaaa gatcttattc cctttaacta ataaaaaaaa ataataaagc atcacttact 29280taaaatcagt tagcaaattt ctgtccagtt tattcagcag cacctccttg ccctcctccc 29340agctctggta ttgcagcttc ctcctggctg caaactttct ccacaatcta aatggaatgt 29400cagtttcctc ctgttcctgt ccatccgcac ccactatctt catgttgttg cagatgaagc 29460gcgcaagacc gtctgaagat accttcaacc ccgtgtatcc atatgacacg gaaaccggtc 29520ctccaactgt gccttttctt actcctccct ttgtatcccc caatgggttt caagagagtc 29580cccctggggt actctctttg cgcctatccg aacctctagt tacctccaat ggcatgcttg 29640cgctcaaaat gggcaacggc ctctctctgg acgaggccgg caaccttacc tcccaaaatg 29700taaccactgt gagcccacct ctcaaaaaaa ccaagtcaaa cataaacctg gaaatatctg 29760cacccctcac agttacctca gaagccctaa ctgtggctgc cgccgcacct ctaatggtcg 29820cgggcaacac actcaccatg caatcacagg ccccgctaac cgtgcacgac tccaaactta 29880gcattgccac ccaaggaccc ctcacagtgt cagaaggaaa gctagccctg caaacatcag 29940gccccctcac caccaccgat agcagtaccc ttactatcac tgcctcaccc cctctaacta 30000ctgccactgg tagcttgggc attgacttga aagagcccat ttatacacaa aatggaaaac

30060taggactaaa gtacggggct cctttgcatg taacagacga cctaaacact ttgaccgtag 30120caactggtcc aggtgtgact attaataata cttccttgca aactaaagtt actggagcct 30180tgggttttga ttcacaaggc aatatgcaac ttaatgtagc aggaggacta aggattgatt 30240ctcaaaacag acgccttata cttgatgtta gttatccgtt tgatgctcaa aaccaactaa 30300atctaagact aggacagggc cctcttttta taaactcagc ccacaacttg gatattaact 30360acaacaaagg cctttacttg tttacagctt caaacaattc caaaaagctt gaggttaacc 30420taagcactgc caaggggttg atgtttgacg ctacagccat agccattaat gcaggagatg 30480ggcttgaatt tggttcacct aatgcaccaa acacaaatcc cctcaaaaca aaaattggcc 30540atggcctaga atttgattca aacaaggcta tggttcctaa actaggaact ggccttagtt 30600ttgacagcac aggtgccatt acagtaggaa acaaaaataa tgataagcta actttgtgga 30660ccacaccagc tccatctcct aactgtagac taaatgcaga gaaagatgct aaactcactt 30720tggtcttaac aaaatgtggc agtcaaatac ttgctacagt ttcagttttg gctgttaaag 30780gcagtttggc tccaatatct ggaacagttc aaagtgctca tcttattata agatttgacg 30840aaaatggagt gctactaaac aattccttcc tggacccaga atattggaac tttagaaatg 30900gagatcttac tgaaggcaca gcctatacaa acgctgttgg atttatgcct aacctatcag 30960cttatccaaa atctcacggt aaaactgcca aaagtaacat tgtcagtcaa gtttacttaa 31020acggagacaa aactaaacct gtaacactaa ccattacact aaacggtaca caggaaacag 31080gagacacaac tccaagtgca tactctatgt cattttcatg ggactggtct ggccacaact 31140acattaatga aatatttgcc acatcctctt acactttttc atacattgcc caagaataaa 31200gaatcgtttg tgttatgttt caacgtgttt atttttcaat tgcagaaaat ttcaagtcat 31260ttttcattca gtagtatagc cccaccacca catagcttat acagatcacc gtaccttaat 31320caaactcaca gaaccctagt attcaacctg ccacctccct cccaacacac agagtacaca 31380gtcctttctc cccggctggc cttaaaaagc atcatatcat gggtaacaga catattctta 31440ggtgttatat tccacacggt ttcctgtcga gccaaacgct catcagtgat attaataaac 31500tccccgggca gctcacttaa gttcatgtcg ctgtccagct gctgagccac aggctgctgt 31560ccaacttgcg gttgcttaac gggcggcgaa ggagaagtcc acgcctacat gggggtagag 31620tcataatcgt gcatcaggat agggcggtgg tgctgcagca gcgcgcgaat aaactgctgc 31680cgccgccgct ccgtcctgca ggaatacaac atggcagtgg tctcctcagc gatgattcgc 31740accgcccgca gcataaggcg ccttgtcctc cgggcacagc agcgcaccct gatctcactt 31800aaatcagcac agtaactgca gcacagcacc acaatattgt tcaaaatccc acagtgcaag 31860gcgctgtatc caaagctcat ggcggggacc acagaaccca cgtggccatc ataccacaag 31920cgcaggtaga ttaagtggcg acccctcata aacacgctgg acataaacat tacctctttt 31980ggcatgttgt aattcaccac ctcccggtac catataaacc tctgattaaa catggcgcca 32040tccaccacca tcctaaacca gctggccaaa acctgcccgc cggctataca ctgcagggaa 32100ccgggactgg aacaatgaca gtggagagcc caggactcgt aaccatggat catcatgctc 32160gtcatgatat caatgttggc acaacacagg cacacgtgca tacacttcct caggattaca 32220agctcctccc gcgttagaac catatcccag ggaacaaccc attcctgaat cagcgtaaat 32280cccacactgc agggaagacc tcgcacgtaa ctcacgttgt gcattgtcaa agtgttacat 32340tcgggcagca gcggatgatc ctccagtatg gtagcgcggg tttctgtctc aaaaggaggt 32400agacgatccc tactgtacgg agtgcgccga gacaaccgag atcgtgttgg tcgtagtgtc 32460atgccaaatg gaacgccgga cgtagtcata tttcctgaag caaaaccagg tgcgggcgtg 32520acaaacagat ctgcgtctcc ggtctcgccg cttagatcgc tctgtgtagt agttgtagta 32580tatccactct ctcaaagcat ccaggcgccc cctggcttcg ggttctatgt aaactccttc 32640atgcgccgct gccctgataa catccaccac cgcagaataa gccacaccca gccaacctac 32700acattcgttc tgcgagtcac acacgggagg agcgggaaga gctggaagaa ccatgttttt 32760ttttttattc caaaagatta tccaaaacct caaaatgaag atctattaag tgaacgcgct 32820cccctccggt ggcgtggtca aactctacag ccaaagaaca gataatggca tttgtaagat 32880gttgcacaat ggcttccaaa aggcaaacgg ccctcacgtc caagtggacg taaaggctaa 32940acccttcagg gtgaatctcc tctataaaca ttccagcacc ttcaaccatg cccaaataat 33000tctcatctcg ccaccttctc aatatatctc taagcaaatc ccgaatatta agtccggcca 33060ttgtaaaaat ctgctccaga gcgccctcca ccttcagcct caagcagcga atcatgattg 33120caaaaattca ggttcctcac agacctgtat aagattcaaa agcggaacat taacaaaaat 33180accgcgatcc cgtaggtccc ttcgcagggc cagctgaaca taatcgtgca ggtctgcacg 33240gaccagcgcg gccacttccc cgccaggaac catgacaaaa gaacccacac tgattatgac 33300acgcatactc ggagctatgc taaccagcgt agccccgatg taagcttgtt gcatgggcgg 33360cgatataaaa tgcaaggtgc tgctcaaaaa atcaggcaaa gcctcgcgca aaaaagaaag 33420cacatcgtag tcatgctcat gcagataaag gcaggtaagc tccggaacca ccacagaaaa 33480agacaccatt tttctctcaa acatgtctgc gggtttctgc ataaacacaa aataaaataa 33540caaaaaaaca tttaaacatt agaagcctgt cttacaacag gaaaaacaac ccttataagc 33600ataagacgga ctacggccat gccggcgtga ccgtaaaaaa actggtcacc gtgattaaaa 33660agcaccaccg acagctcctc ggtcatgtcc ggagtcataa tgtaagactc ggtaaacaca 33720tcaggttgat tcacatcggt cagtgctaaa aagcgaccga aatagcccgg gggaatacat 33780acccgcaggc gtagagacaa cattacagcc cccataggag gtataacaaa attaatagga 33840gagaaaaaca cataaacacc tgaaaaaccc tcctgcctag gcaaaatagc accctcccgc 33900tccagaacaa catacagcgc ttccacagcg gcagccataa cagtcagcct taccagtaaa 33960aaagaaaacc tattaaaaaa acaccactcg acacggcacc agctcaatca gtcacagtgt 34020aaaaaagggc caagtgcaga gcgagtatat ataggactaa aaaatgacgt aacggttaaa 34080gtccacaaaa aacacccaga aaaccgcacg cgaacctacg cccagaaacg aaagccaaaa 34140aacccacaac ttcctcaaat cgtcacttcc gttttcccac gttacgtcac ttcccatttt 34200aagaaaacta caattcccaa cacatacaag ttactccgcc ctaaaaccta cgtcacccgc 34260cccgttccca cgccccgcgc cacgtcacaa actccacccc ctcattatca tattggcttc 34320aatccaaaat aaggtatatt attgatgatg 343502590DNAArtificial sequenceTNFR portion of the TNFR-Fas chimera (Fas-c) 2atgggcctct ccaccgtgcc tgacctgctg ctgccgctgg tgctcctgga gctgttggtg 60ggaatatacc cctcaggggt tattggactg gtccctcacc taggggacag ggagaagaga 120gatagtgtgt gtccccaagg aaaatatatc caccctcaaa ataattcgat ttgctgtacc 180aagtgccaca aaggaaccta cttgtacaat gactgtccag gcccggggca ggatacggac 240tgcagggagt gtgagagcgg ctccttcacc gcttcagaaa accacctcag acactgcctc 300agctgctcca aatgccgaaa ggaaatgggt caggtggaga tctcttcttg cacagtggac 360cgggacaccg tgtgtggctg caggaagaac cagtaccggc attattggag tgaaaacctt 420ttccagtgct tcaattgcag cctctgcctc aatgggaccg tgcacctctc ctgccaggag 480aaacagaaca ccgtgtgcac ctgccatgca ggtttctttc taagagaaaa cgagtgtgtc 540tcctgtagta actgtaagaa aagcctggag tgcacgaagt tgtgcctacc 5903511DNAArtificial sequenceFas portion of the TNFR-Fas chimera (Fas-c) 3aagcttagga tccagatcta acttggggtg gctttgtctt cttcttttgc caattccact 60aattgtttgg gtgaagagaa aggaagtaca gaaaacatgc agaaagcaca gaaaggaaaa 120ccaaggttct catgaatctc caaccttaaa tcctgaaaca gtggcaataa atttatctga 180tgttgacttg agtaaatata tcaccactat tgctggagtc atgacactaa gtcaagttaa 240aggctttgtt cgaaagaatg gtgtcaatga agccaaaata gatgagatca agaatgacaa 300tgtccaagac acagcagaac agaaagttca actgcttcgt aattggcatc aacttcatgg 360aaagaaagaa gcgtatgaca cattgattaa agatctcaaa aaagccaatc tttgtactct 420tgcagagaaa attcagacta tcatcctcaa ggacattact agtgactcag aaaattcaaa 480cttcagaaat gaaatccaaa gcttggtcta g 51141101DNAArtificial sequenceTNFR1-Fas chimera (Fas-c) coding sequence 4atgggcctct ccaccgtgcc tgacctgctg ctgccgctgg tgctcctgga gctgttggtg 60ggaatatacc cctcaggggt tattggactg gtccctcacc taggggacag ggagaagaga 120gatagtgtgt gtccccaagg aaaatatatc caccctcaaa ataattcgat ttgctgtacc 180aagtgccaca aaggaaccta cttgtacaat gactgtccag gcccggggca ggatacggac 240tgcagggagt gtgagagcgg ctccttcacc gcttcagaaa accacctcag acactgcctc 300agctgctcca aatgccgaaa ggaaatgggt caggtggaga tctcttcttg cacagtggac 360cgggacaccg tgtgtggctg caggaagaac cagtaccggc attattggag tgaaaacctt 420ttccagtgct tcaattgcag cctctgcctc aatgggaccg tgcacctctc ctgccaggag 480aaacagaaca ccgtgtgcac ctgccatgca ggtttctttc taagagaaaa cgagtgtgtc 540tcctgtagta actgtaagaa aagcctggag tgcacgaagt tgtgcctacc aagcttagga 600tccagatcta acttggggtg gctttgtctt cttcttttgc caattccact aattgtttgg 660gtgaagagaa aggaagtaca gaaaacatgc agaaagcaca gaaaggaaaa ccaaggttct 720catgaatctc caaccttaaa tcctgaaaca gtggcaataa atttatctga tgttgacttg 780agtaaatata tcaccactat tgctggagtc atgacactaa gtcaagttaa aggctttgtt 840cgaaagaatg gtgtcaatga agccaaaata gatgagatca agaatgacaa tgtccaagac 900acagcagaac agaaagttca actgcttcgt aattggcatc aacttcatgg aaagaaagaa 960gcgtatgaca cattgattaa agatctcaaa aaagccaatc tttgtactct tgcagagaaa 1020attcagacta tcatcctcaa ggacattact agtgactcag aaaattcaaa cttcagaaat 1080gaaatccaaa gcttggtcta g 110156DNAArtificial sequenceHypoxia responsive element - E-box 5gcacgt 6644DNAArtificial sequenceMurine endothelial specific enhancer element 6gtacttcata cttttcattc caatggggtg actttgcttc tgga 447143DNAArtificial sequenceA triplicate copy of a murine enhancer sequence originated from the PPE-1 promoter 7gtacttcata cttttcattc caatggggtg actttgcttc tggagggtga ctttgcttct 60ggagccagta cttcatactt ttcattgtac ttcatacttt tcattccaat ggggtgactt 120tgcttctgga ggctagctgc cag 143847DNAArtificial sequenceEDC fragment 8ctggagggtg actttgcttc tggagccagt acttcatact tttcatt 47936460DNAArtificial sequencePPE-1-3X-FasC virl construct including repeat sequences 9catcatcaat aatatacctt attttggatt gaagccaata tgataatgag ggggtggagt 60ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg tagtagtgtg gcggaagtgt 120gatgttgcaa gtgtggcgga acacatgtaa gcgacggatg tggcaaaagt gacgtttttg 180gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg gttttaggcg gatgttgtag 240taaatttggg cgtaaccgag taagatttgg ccattttcgc gggaaaactg aataagagga 300agtgaaatct gaataatttt gtgttactca tagcgcgtaa tatttgtcta gggccgcggg 360gactttgacc gtttacgtgg agactcgccc aggtgttttt ctcaggtgtt ttccgcgttc 420cgggtcaaag ttggcgtttt attattatag tcagtacgta cgtgtacttc tgatcggcga 480tactagggag ataaggatgt acctgacaaa accacattgt tgttgttatc attattattt 540agttttcctt ccttgctaac tcctgacgga atctttctca cctcaaatgc gaagtacttt 600agtttagaaa agacttggtg gaaggggtgg tggtggaaaa gtagggtgat cttccaaact 660aatctggttc cccgcccgcc ccagtagctg ggattcaaga gcgaagagtg gggatcgtcc 720ccttgtttga tcagaaagac ataaaaggaa aatcaagtga acaatgatca gccccacctc 780caccccaccc ccctgcgcgc gcacaataca atctatttaa ttgtacttca tacttttcat 840tccaatgggg tgactttgct tctggagaaa ctcttgattc ttgaactctg gggctggcag 900ctagcctcca gaagcaaagt caccccattg gaatgaaaag tatgaagtac aatgaaaagt 960atgaagtact ggctccagaa gcaaagtcac cctccagaag caaagtcacc ccattggaat 1020gaaaagtatg aagtacgcta gcaaaagggg aagcgggctg ctgctctctg caggttctgc 1080agcggtctct gtctagtggg tgttttcttt ttcttagccc tgcccctgga ttgtcagacg 1140gcgggcgtct gcctctgaag ttagccgtga tttcctctag agccgggtct tatctctggc 1200tgcacgttgc ctgtgggtga ctaatcacac aataacattg tttagggctg gaataaagtc 1260agagctgttt acccccactc tataggggtt caatataaaa aggcggcgga gaactgtccg 1320agtcagaagc gttcctgcac cggcgctgag agcctgaccc ggtctgctcc gctgtccttg 1380cgcgctgcct cccggctgcc cgcgacgctt tcgccccagt ggaagggcca cttgctgcgg 1440ccgctaattc tgcagatcgg gatccggcat gggcctctcc accgtgcctg acctgctgct 1500gccgctggtg ctcctggagc tgttggtggg aatatacccc tcaggggtta ttggactggt 1560ccctcaccta ggggacaggg agaagagaga tagtgtgtgt ccccaaggaa aatatatcca 1620ccctcaaaat aattcgattt gctgtaccaa gtgccacaaa ggaacctact tgtacaatga 1680ctgtccaggc ccggggcagg atacggactg cagggagtgt gagagcggct ccttcaccgc 1740ttcagaaaac cacctcagac actgcctcag ctgctccaaa tgccgaaagg aaatgggtca 1800ggtggagatc tcttcttgca cagtggaccg ggacaccgtg tgtggctgca ggaagaacca 1860gtaccggcat tattggagtg aaaacctttt ccagtgcttc aattgcagcc tctgcctcaa 1920tgggaccgtg cacctctcct gccaggagaa acagaacacc gtgtgcacct gccatgcagg 1980tttctttcta agagaaaacg agtgtgtctc ctgtagtaac tgtaagaaaa gcctggagtg 2040cacgaagttg tgcctaccaa gcttaggatc cagatctaac ttggggtggc tttgtcttct 2100tcttttgcca attccactaa ttgtttgggt gaagagaaag gaagtacaga aaacatgcag 2160aaagcacaga aaggaaaacc aaggttctca tgaatctcca accttaaatc ctgaaacagt 2220ggcaataaat ttatctgatg ttgacttgag taaatatatc accactattg ctggagtcat 2280gacactaagt caagttaaag gctttgttcg aaagaatggt gtcaatgaag ccaaaataga 2340tgagatcaag aatgacaatg tccaagacac agcagaacag aaagttcaac tgcttcgtaa 2400ttggcatcaa cttcatggaa agaaagaagc gtatgacaca ttgattaaag atctcaaaaa 2460agccaatctt tgtactcttg cagagaaaat tcagactatc atcctcaagg acattactag 2520tgactcagaa aattcaaact tcagaaatga aatccaaagc ttggtctagc tcgagcatgc 2580atctaggcgg ccgcatggca gaaattcgcg aattcgctag cgttaacgga tcctctagac 2640gagatccgaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 2700atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 2760atgtatctta tcatgtctag atctgtactg aaatgtgtgg gcgtggctta agggtgggaa 2820agaatatata aggtgggggt cttatgtagt tttgtatctg ttttgcagca gccgccgccg 2880ccatgagcac caactcgttt gatggaagca ttgtgagctc atatttgaca acgcgcatgc 2940ccccatgggc cggggtgcgt cagaatgtga tgggctccag cattgatggt cgccccgtcc 3000tgcccgcaaa ctctactacc ttgacctacg agaccgtgtc tggaacgccg ttggagactg 3060cagcctccgc cgccgcttca gccgctgcag ccaccgcccg cgggattgtg actgactttg 3120ctttcctgag cccgcttgca agcagtgcag cttcccgttc atccgcccgc gatgacaagt 3180tgacggctct tttggcacaa ttggattctt tgacccggga acttaatgtc gtttctcagc 3240agctgttgga tctgcgccag caggtttctg ccctgaaggc ttcctcccct cccaatgcgg 3300tttaaaacat aaataaaaaa ccagactctg tttggatttg gatcaagcaa gtgtcttgct 3360gtctttattt aggggttttg cgcgcgcggt aggcccggga ccagcggtct cggtcgttga 3420gggtcctgtg tattttttcc aggacgtggt aaaggtgact ctggatgttc agatacatgg 3480gcataagccc gtctctgggg tggaggtagc accactgcag agcttcatgc tgcggggtgg 3540tgttgtagat gatccagtcg tagcaggagc gctgggcgtg gtgcctaaaa atgtctttca 3600gtagcaagct gattgccagg ggcaggccct tggtgtaagt gtttacaaag cggttaagct 3660gggatgggtg catacgtggg gatatgagat gcatcttgga ctgtattttt aggttggcta 3720tgttcccagc catatccctc cggggattca tgttgtgcag aaccaccagc acagtgtatc 3780cggtgcactt gggaaatttg tcatgtagct tagaaggaaa tgcgtggaag aacttggaga 3840cgcccttgtg acctccaaga ttttccatgc attcgtccat aatgatggca atgggcccac 3900gggcggcggc ctgggcgaag atatttctgg gatcactaac gtcatagttg tgttccagga 3960tgagatcgtc ataggccatt tttacaaagc gcgggcggag ggtgccagac tgcggtataa 4020tggttccatc cggcccaggg gcgtagttac cctcacagat ttaagggtgg gaaagaatat 4080ataaggtggg ggtcttatgt agttttgtat ctgttttgca gcagccgccg ccgccatgag 4140caccaactcg tttgatggaa gcattgtgag ctcatatttg acaacgcgca tgcccccatg 4200ggccggggtg cgtcagaatg tgatgggctc cagcattgat ggtcgccccg tcctgcccgc 4260aaactctact accttgacct acgagaccgt gtctggaacg ccgttggaga ctgcagcctc 4320cgccgccgct tcagccgctg cagccaccgc ccgcgggatt gtgactgact ttgctttcct 4380gagcccgctt gcaagcagtg cagcttcccg ttcatccgcc cgcgatgaca agttgacggc 4440tcttttggca caattggatt ctttgacccg ggaacttaat gtcgtttctc agcagctgtt 4500ggatctgcgc cagcaggttt ctgccctgaa ggcttcctcc cctcccaatg cggtttaaaa 4560cataaataaa aaaccagact ctgtttggat ttggatcaag caagtgtctt gctgtcttta 4620tttaggggtt ttgcgcgcgc ggtaggcccg ggaccagcgg tctcggtcgt tgagggtcct 4680gtgtattttt tccaggacgt ggtaaaggtg actctggatg ttcagataca tgggcataag 4740cccgtctctg gggtggaggt agcaccactg cagagcttca tgctgcgggg tggtgttgta 4800gatgatccag tcgtagcagg agcgctgggc gtggtgccta aaaatgtctt tcagtagcaa 4860gctgattgcc aggggcaggc ccttggtgta agtgtttaca aagcggttaa gctgggatgg 4920gtgcatacgt ggggatatga gatgcatctt ggactgtatt tttaggttgg ctatgttccc 4980agccatatcc ctccggggat tcatgttgtg cagaaccacc agcacagtgt atccggtgca 5040cttgggaaat ttgtcatgta gcttagaagg aaatgcgtgg aagaacttgg agacgccctt 5100gtgacctcca agattttcca tgcattcgtc cataatgatg gcaatgggcc cacgggcggc 5160ggcctgggcg aagatatttc tgggatcact aacgtcatag ttgtgttcca ggatgagatc 5220gtcataggcc atttttacaa agcgcgggcg gagggtgcca gactgcggta taatggttcc 5280atccggccca ggggcgtagt taccctcaca gatttgcatt tcccacgctt tgagttcaga 5340tggggggatc atgtctacct gcggggcgat gaagaaaacg gtttccgggg taggggagat 5400cagctgggaa gaaagcaggt tcctgagcag ctgcgactta ccgcagccgg tgggcccgta 5460aatcacacct attaccggct gcaactggta gttaagagag ctgcagctgc cgtcatccct 5520gagcaggggg gccacttcgt taagcatgtc cctgactcgc atgttttccc tgaccaaatc 5580cgccagaagg cgctcgccgc ccagcgatag cagttcttgc aaggaagcaa agtttttcaa 5640cggtttgaga ccgtccgccg taggcatgct tttgagcgtt tgaccaagca gttccaggcg 5700gtcccacagc tcggtcacct gctctacggc atctcgatcc agcatatctc ctcgtttcgc 5760gggttggggc ggctttcgct gtacggcagt agtcggtgct cgtccagacg ggccagggtc 5820atgtctttcc acgggcgcag ggtcctcgtc agcgtagtct gggtcacggt gaaggggtgc 5880gctccgggct gcgcgctggc cagggtgcgc ttgaggctgg tcctgctggt gctgaagcgc 5940tgccggtctt cgccctgcgc gtcggccagg tagcatttga ccatggtgtc atagtccagc 6000ccctccgcgg cgtggccctt ggcgcgcagc ttgcccttgg aggaggcgcc gcacgagggg 6060cagtgcagac ttttgagggc gtagagcttg ggcgcgagaa ataccgattc cggggagtag 6120gcatccgcgc cgcaggcccc gcagacggtc tcgcattcca cgagccaggt gagctctggc 6180cgttcggggt caaaaaccag gtttccccca tgctttttga tgcgtttctt acctctggtt 6240tccatgagcc ggtgtccacg ctcggtgacg aaaaggctgt ccgtgtcccc gtatacagac 6300ttgagaggcc tgtcctcgag cggtgttccg cggtcctcct cgtatagaaa ctcggaccac 6360tctgagacaa aggctcgcgt ccaggccagc acgaaggagg ctaagtggga ggggtagcgg 6420tcgttgtcca ctagggggtc cactcgctcc agggtgtgaa gacacatgtc gccctcttcg 6480gcatcaagga aggtgattgg tttgtaggtg taggccacgt gaccgggtgt tcctgaaggg 6540gggctataaa agggggtggg ggcgcgttcg tcctcactct cttccgcatc gctgtctgcg 6600agggccagct gttggggtga gtactccctc tgaaaagcgg gcatgacttc tgcgctaaga 6660ttgtcagttt ccaaaaacga ggaggatttg atattcacct ggcccgcggt gatgcctttg 6720agggtggccg catccatctg gtcagaaaag acaatctttt tgttgtcaag cttggtggca 6780aacgacccgt agagggcgtt ggacagcaac ttggcgatgg agcgcagggt ttggtttttg 6840tcgcgatcgg cgcgctcctt ggccgcgatg tttagctgca cgtattcgcg cgcaacgcac 6900cgccattcgg gaaagacggt ggtgcgctcg tcgggcacca ggtgcacgcg ccaaccgcgg 6960ttgtgcaggg tgacaaggtc aacgctggtg gctacctctc cgcgtaggcg ctcgttggtc 7020cagcagaggc ggccgccctt gcgcgagcag aatggcggta gggggtctag ctgcgtctcg 7080tccggggggt ctgcgtccac ggtaaagacc ccgggcagca ggcgcgcgtc gaagtagtct 7140atcttgcatc cttgcaagtc tagcgcctgc tgccatgcgc gggcggcaag cgcgcgctcg 7200tatgggttga gtgggggacc ccatggcatg gggtgggtga gcgcggaggc gtacatgccg 7260caaatgtcgt aaacgtagag gggctctctg agtattccaa gatatgtagg gtagcatctt 7320ccaccgcgga tgctggcgcg cacgtaatcg tatagttcgt gcgagggagc gaggaggtcg 7380ggaccgaggt tgctacgggc gggctgctct gctcggaaga ctatctgcct gaagatggca 7440tgtgagttgg atgatatggt tggacgctgg aagacgttga agctggcgtc tgtgagacct 7500accgcgtcac gcacgaagga ggcgtaggag tcgcgcagct tgttgaccag ctcggcggtg 7560acctgcacgt ctagggcgca

gtagtccagg gtttccttga tgatgtcata cttatcctgt 7620cccttttttt tccacagctc gcggttgagg acaaactctt cgcggtcttt ccagtactct 7680tggatcggaa acccgtcggc ctccgaacgg taagagccta gcatgtagaa ctggttgacg 7740gcctggtagg cgcagcatcc cttttctacg ggtagcgcgt atgcctgcgc ggccttccgg 7800agcgaggtgt gggtgagcgc aaaggtgtcc ctgaccatga ctttgaggta ctggtatttg 7860aagtcagtgt cgtcgcatcc gccctgctcc cagagcaaaa agtccgtgcg ctttttggaa 7920cgcggatttg gcagggcgaa ggtgacatcg ttgaagagta tctttcccgc gcgaggcata 7980aagttgcgtg tgatgcggaa gggtcccggc acctcggaac ggttgttaat tacctgggcg 8040gcgagcacga tctcgtcaaa gccgttgatg ttgtggccca caatgtaaag ttccaagaag 8100cgcgggatgc ccttgatgga aggcaatttt ttaagttcct cgtaggtgag ctcttcaggg 8160gagctgagcc cgtgctctga aagggcccag tctgcaagat gagggttgga agcgacgaat 8220gagctccaca ggtcacgggc cattagcatt tgcaggtggt cgcgaaaggt cctaaactgg 8280cgacctatgg ccattttttc tggggtgatg cagtagaagg taagcgggtc ttgttcccag 8340cggtcccatc caaggttcgc ggctaggtct cgcgcggcag tcactagagg ctcatctccg 8400ccgaacttca tgaccagcat gaagggcacg agctgcttcc caaaggcccc catccaagta 8460taggtctcta catcgtaggt gacaaagaga cgctcggtgc gaggatgcga gccgatcggg 8520aagaactgga tctcccgcca ccaattggag gagtggctat tgatgtggtg aaagtagaag 8580tccctgcgac gggccgaaca ctcgtgctgg cttttgtaaa aacgtgcgca gtactggcag 8640cggtgcacgg gctgtacatc ctgcacgagg ttgacctgac gaccgcgcac aaggaagcag 8700agtgggaatt tgagcccctc gcctggcggg tttggctggt ggtcttctac ttcggctgct 8760tgtccttgac cgtctggctg ctcgagggga gttacggtgg atcggaccac cacgccgcgc 8820gagcccaaag tccagatgtc cgcgcgcggc ggtcggagct tgatgacaac atcgcgcaga 8880tgggagctgt ccatggtctg gagctcccgc ggcgtcaggt caggcgggag ctcctgcagg 8940tttacctcgc atagacgggt cagggcgcgg gctagatcca ggtgatacct aatttccagg 9000ggctggttgg tggcggcgtc gatggcttgc aagaggccgc atccccgcgg cgcgactacg 9060gtaccgcgcg gcgggcggtg ggccgcgggg gtgtccttgg atgatgcatc taaaagcggt 9120gacgcgggcg agcccccgga ggtagggggg gctccggacc cgccgggaga gggggcaggg 9180gcacgtcggc gccgcgcgcg ggcaggagct ggtgctgcgc gcgtaggttg ctggcgaacg 9240cgacgacgcg gcggttgatc tcctgaatct ggcgcctctg cgtgaagacg acgggcccgg 9300tgagcttgaa cctgaaagag agttcgacag aatcaatttc ggtgtcgttg acggcggcct 9360ggcgcaaaat ctcctgcacg tctcctgagt tgtcttgata ggcgatctcg gccatgaact 9420gctcgatctc ttcctcctgg agatctccgc gtccggctcg ctccacggtg gcggcgaggt 9480cgttggaaat gcgggccatg agctgcgaga aggcgttgag gcctccctcg ttccagacgc 9540ggctgtagac cacgccccct tcggcatcgc gggcgcgcat gaccacctgc gcgagattga 9600gctccacgtg ccgggcgaag acggcgtagt ttcgcaggcg ctgaaagagg tagttgaggg 9660tggtggcggt gtgttctgcc acgaagaagt acataaccca gcgtcgcaac gtggattcgt 9720tgatatcccc caaggcctca aggcgctcca tggcctcgta gaagtccacg gcgaagttga 9780aaaactggga gttgcgcgcc gacacggtta actcctcctc cagaagacgg atgagctcgg 9840cgacagtgtc gcgcacctcg cgctcaaagg ctacaggggc ctcttcttct tcttcaatct 9900cctcttccat aagggcctcc ccttcttctt cttctggcgg cggtggggga ggggggacac 9960ggcggcgacg acggcgcacc gggaggcggt cgacaaagcg ctcgatcatc tccccgcggc 10020gacggcgcat ggtctcggtg acggcgcggc cgttctcgcg ggggcgcagt tggaagacgc 10080cgcccgtcat gtcccggtta tgggttggcg gggggctgcc atgcggcagg gatacggcgc 10140taacgatgca tctcaacaat tgttgtgtag gtactccgcc gccgagggac ctgagcgagt 10200ccgcatcgac cggatcggaa aacctctcga gaaaggcgtc taaccagtca cagtcgcaag 10260gtaggctgag caccgtggcg ggcggcagcg ggcggcggtc ggggttgttt ctggcggagg 10320tgctgctgat gatgtaatta aagtaggcgg tcttgagacg gcggatggtc gacagaagca 10380ccatgtcctt gggtccggcc tgctgaatgc gcaggcggtc ggccatgccc caggcttcgt 10440tttgacatcg gcgcaggtct ttgtagtagt cttgcatgag cctttctacc ggcacttctt 10500cttctccttc ctcttgtcct gcatctcttg catctatcgc tgcggcggcg gcggagtttg 10560gccgtaggtg gcgccctctt cctcccatgc gtgtgacccc gaagcccctc atcggctgaa 10620gcagggctag gtcggcgaca acgcgctcgg ctaatatggc ctgctgcacc tgcgtgaggg 10680tagactggaa gtcatccatg tccacaaagc ggtggtatgc gcccgtgttg atggtgtaag 10740tgcagttggc cataacggac cagttaacgg tctggtgacc cggctgcgag agctcggtgt 10800acctgagacg cgagtaagcc ctcgagtcaa atacgtagtc gttgcaagtc cgcaccaggt 10860actggtatcc caccaaaaag tgcggcggcg gctggcggta gaggggccag cgtagggtgg 10920ccggggctcc gggggcgaga tcttccaaca taaggcgatg atatccgtag atgtacctgg 10980acatccaggt gatgccggcg gcggtggtgg aggcgcgcgg aaagtcgcgg acgcggttcc 11040agatgttgcg cagcggcaaa aagtgctcca tggtcgggac gctctggccg gtcaggcgcg 11100cgcaatcgtt gacgctctag accgtgcaaa aggagagcct gtaagcgggc actcttccgt 11160ggtctggtgg ataaattcgc aagggtatca tggcggacga ccggggttcg agccccgtat 11220ccggccgtcc gccgtgatcc atgcggttac cgcccgcgtg tcgaacccag gtgtgcgacg 11280tcagacaacg ggggagtgct ccttttggct tccttccagg cgcggcggct gctgcgctag 11340cttttttggc cactggccgc gcgcagcgta agcggttagg ctggaaagcg aaagcattaa 11400gtggctcgct ccctgtagcc ggagggttat tttccaaggg ttgagtcgcg ggacccccgg 11460ttcgagtctc ggaccggccg gactgcggcg aacgggggtt tgcctccccg tcatgcaaga 11520ccccgcttgc aaattcctcc ggaaacaggg acgagcccct tttttgcttt tcccagatgc 11580atccggtgct gcggcagatg cgcccccctc ctcagcagcg gcaagagcaa gagcagcggc 11640agacatgcag ggcaccctcc cctcctccta ccgcgtcagg aggggcgaca tccgcggttg 11700acgcggcagc agatggtgat tacgaacccc cgcggcgccg ggcccggcac tacctggact 11760tggaggaggg cgagggcctg gcgcggctag gagcgccctc tcctgagcgg cacccaaggg 11820tgcagctgaa gcgtgatacg cgtgaggcgt acgtgccgcg gcagaacctg tttcgcgacc 11880gcgagggaga ggagcccgag gagatgcggg atcgaaagtt ccacgcaggg cgcgagctgc 11940ggcatggcct gaatcgcgag cggttgctgc gcgaggagga ctttgagccc gacgcgcgaa 12000ccgggattag tcccgcgcgc gcacacgtgg cggccgccga cctggtaacc gcatacgagc 12060agacggtgaa ccaggagatt aactttcaaa aaagctttaa caaccacgtg cgtacgcttg 12120tggcgcgcga ggaggtggct ataggactga tgcatctgtg ggactttgta agcgcgctgg 12180agcaaaaccc aaatagcaag ccgctcatgg cgcagctgtt ccttatagtg cagcacagca 12240gggacaacga ggcattcagg gatgcgctgc taaacatagt agagcccgag ggccgctggc 12300tgctcgattt gataaacatc ctgcagagca tagtggtgca ggagcgcagc ttgagcctgg 12360ctgacaaggt ggccgccatc aactattcca tgcttagcct gggcaagttt tacgcccgca 12420agatatacca taccccttac gttcccatag acaaggaggt aaagatcgag gggttctaca 12480tgcgcatggc gctgaaggtg cttaccttga gcgacgacct gggcgtttat cgcaacgagc 12540gcatccacaa ggccgtgagc gtgagccggc ggcgcgagct cagcgaccgc gagctgatgc 12600acagcctgca aagggccctg gctggcacgg gcagcggcga tagagaggcc gagtcctact 12660ttgacgcggg cgctgacctg cgctgggccc caagccgacg cgccctggag gcagctgggg 12720ccggacctgg gctggcggtg gcacccgcgc gcgctggcaa cgtcggcggc gtggaggaat 12780atgacgagga cgatgagtac gagccagagg acggcgagta ctaagcggtg atgtttctga 12840tcagatgatg caagacgcaa cggacccggc ggtgcgggcg gcgctgcaga gccagccgtc 12900cggccttaac tccacggacg actggcgcca ggtcatggac cgcatcatgt cgctgactgc 12960gcgcaatcct gacgcgttcc ggcagcagcc gcaggccaac cggctctccg caattctgga 13020agcggtggtc ccggcgcgcg caaaccccac gcacgagaag gtgctggcga tcgtaaacgc 13080gctggccgaa aacagggcca tccggcccga cgaggccggc ctggtctacg acgcgctgct 13140tcagcgcgtg gctcgttaca acagcggcaa cgtgcagacc aacctggacc ggctggtggg 13200ggatgtgcgc gaggccgtgg cgcagcgtga gcgcgcgcag cagcagggca acctgggctc 13260catggttgca ctaaacgcct tcctgagtac acagcccgcc aacgtgccgc ggggacagga 13320ggactacacc aactttgtga gcgcactgcg gctaatggtg actgagacac cgcaaagtga 13380ggtgtaccag tctgggccag actatttttt ccagaccagt agacaaggcc tgcagaccgt 13440aaacctgagc caggctttca aaaacttgca ggggctgtgg ggggtgcggg ctcccacagg 13500cgaccgcgcg accgtgtcta gcttgctgac gcccaactcg cgcctgttgc tgctgctaat 13560agcgcccttc acggacagtg gcagcgtgtc ccgggacaca tacctaggtc acttgctgac 13620actgtaccgc gaggccatag gtcaggcgca tgtggacgag catactttcc aggagattac 13680aagtgtcagc cgcgcgctgg ggcaggagga cacgggcagc ctggaggcaa ccctaaacta 13740cctgctgacc aaccggcggc agaagatccc ctcgttgcac agtttaaaca gcgaggagga 13800gcgcattttg cgctacgtgc agcagagcgt gagccttaac ctgatgcgcg acggggtaac 13860gcccagcgtg gcgctggaca tgaccgcgcg caacatggaa ccgggcatgt atgcctcaaa 13920ccggccgttt atcaaccgcc taatggacta cttgcatcgc gcggccgccg tgaaccccga 13980gtatttcacc aatgccatct tgaacccgca ctggctaccg ccccctggtt tctacaccgg 14040gggattcgag gtgcccgagg gtaacgatgg attcctctgg gacgacatag acgacagcgt 14100gttttccccg caaccgcaga ccctgctaga gttgcaacag cgcgagcagg cagaggcggc 14160gctgcgaaag gaaagcttcc gcaggccaag cagcttgtcc gatctaggcg ctgcggcccc 14220gcggtcagat gctagtagcc catttccaag cttgataggg tctcttacca gcactcgcac 14280cacccgcccg cgcctgctgg gcgaggagga gtacctaaac aactcgctgc tgcagccgca 14340gcgcgaaaaa aacctgcctc cggcatttcc caacaacggg atagagagcc tagtggacaa 14400gatgagtaga tggaagacgt acgcgcagga gcacagggac gtgccaggcc cgcgcccgcc 14460cacccgtcgt caaaggcacg accgtcagcg gggtctggtg tgggaggacg atgactcggc 14520agacgacagc agcgtcctgg atttgggagg gagtggcaac ccgtttgcgc accttcgccc 14580caggctgggg agaatgtttt aaaaaaaaaa aaagcatgat gcaaaataaa aaactcacca 14640aggccatggc accgagcgtt ggttttcttg tattcccctt agtatgcggc gcgcggcgat 14700gtatgaggaa ggtcctcctc cctcctacga gagtgtggtg agcgcggcgc cagtggcggc 14760ggcgctgggt tctcccttcg atgctcccct ggacccgccg tttgtgcctc cgcggtacct 14820gcggcctacc ggggggagaa acagcatccg ttactctgag ttggcacccc tattcgacac 14880cacccgtgtg tacctggtgg acaacaagtc aacggatgtg gcatccctga actaccagaa 14940cgaccacagc aactttctga ccacggtcat tcaaaacaat gactacagcc cgggggaggc 15000aagcacacag accatcaatc ttgacgaccg gtcgcactgg ggcggcgacc tgaaaaccat 15060cctgcatacc aacatgccaa atgtgaacga gttcatgttt accaataagt ttaaggcgcg 15120ggtgatggtg tcgcgcttgc ctactaagga caatcaggtg gagctgaaat acgagtgggt 15180ggagttcacg ctgcccgagg gcaactactc cgagaccatg accatagacc ttatgaacaa 15240cgcgatcgtg gagcactact tgaaagtggg cagacagaac ggggttctgg aaagcgacat 15300cggggtaaag tttgacaccc gcaacttcag actggggttt gaccccgtca ctggtcttgt 15360catgcctggg gtatatacaa acgaagcctt ccatccagac atcattttgc tgccaggatg 15420cggggtggac ttcacccaca gccgcctgag caacttgttg ggcatccgca agcggcaacc 15480cttccaggag ggctttagga tcacctacga tgatctggag ggtggtaaca ttcccgcact 15540gttggatgtg gacgcctacc aggcgagctt gaaagatgac accgaacagg gcgggggtgg 15600cgcaggcggc agcaacagca gtggcagcgg cgcggaagag aactccaacg cggcagccgc 15660ggcaatgcag ccggtggagg acatgaacga tcatgccatt cgcggcgaca cctttgccac 15720acgggctgag gagaagcgcg ctgaggccga agcagcggcc gaagctgccg cccccgctgc 15780gcaacccgag gtcgagaagc ctcagaagaa accggtgatc aaacccctga cagaggacag 15840caagaaacgc agttacaacc taataagcaa tgacagcacc ttcacccagt accgcagctg 15900gtaccttgca tacaactacg gcgaccctca gaccggaatc cgctcatgga ccctgctttg 15960cactcctgac gtaacctgcg gctcggagca ggtctactgg tcgttgccag acatgatgca 16020agaccccgtg accttccgct ccacgcgcca gatcagcaac tttccggtgg tgggcgccga 16080gctgttgccc gtgcactcca agagcttcta caacgaccag gccgtctact cccaactcat 16140ccgccagttt acctctctga cccacgtgtt caatcgcttt cccgagaacc agattttggc 16200gcgcccgcca gcccccacca tcaccaccgt cagtgaaaac gttcctgctc tcacagatca 16260cgggacgcta ccgctgcgca acagcatcgg aggagtccag cgagtgacca ttactgacgc 16320cagacgccgc acctgcccct acgtttacaa ggccctgggc atagtctcgc cgcgcgtcct 16380atcgagccgc actttttgag caagcatgtc catccttata tcgcccagca ataacacagg 16440ctggggcctg cgcttcccaa gcaagatgtt tggcggggcc aagaagcgct ccgaccaaca 16500cccagtgcgc gtgcgcgggc actaccgcgc gccctggggc gcgcacaaac gcggccgcac 16560tgggcgcacc accgtcgatg acgccatcga cgcggtggtg gaggaggcgc gcaactacac 16620gcccacgccg ccaccagtgt ccacagtgga cgcggccatt cagaccgtgg tgcgcggagc 16680ccggcgctat gctaaaatga agagacggcg gaggcgcgta gcacgtcgcc accgccgccg 16740acccggcact gccgcccaac gcgcggcggc ggccctgctt aaccgcgcac gtcgcaccgg 16800ccgacgggcg gccatgcggg ccgctcgaag gctggccgcg ggtattgtca ctgtgccccc 16860caggtccagg cgacgagcgg ccgccgcagc agccgcggcc attagtgcta tgactcaggg 16920tcgcaggggc aacgtgtatt gggtgcgcga ctcggttagc ggcctgcgcg tgcccgtgcg 16980cacccgcccc ccgcgcaact agattgcaag aaaaaactac ttagactcgt actgttgtat 17040gtatccagcg gcggcggcgc gcaacgaagc tatgtccaag cgcaaaatca aagaagagat 17100gctccaggtc atcgcgccgg agatctatgg ccccccgaag aaggaagagc aggattacaa 17160gccccgaaag ctaaagcggg tcaaaaagaa aaagaaagat gatgatgatg aacttgacga 17220cgaggtggaa ctgctgcacg ctaccgcgcc caggcgacgg gtacagtgga aaggtcgacg 17280cgtaaaacgt gttttgcgac ccggcaccac cgtagtcttt acgcccggtg agcgctccac 17340ccgcacctac aagcgcgtgt atgatgaggt gtacggcgac gaggacctgc ttgagcaggc 17400caacgagcgc ctcggggagt ttgcctacgg aaagcggcat aaggacatgc tggcgttgcc 17460gctggacgag ggcaacccaa cacctagcct aaagcccgta acactgcagc aggtgctgcc 17520cgcgcttgca ccgtccgaag aaaagcgcgg cctaaagcgc gagtctggtg acttggcacc 17580caccgtgcag ctgatggtac ccaagcgcca gcgactggaa gatgtcttgg aaaaaatgac 17640cgtggaacct gggctggagc ccgaggtccg cgtgcggcca atcaagcagg tggcgccggg 17700actgggcgtg cagaccgtgg acgttcagat acccactacc agtagcacca gtattgccac 17760cgccacagag ggcatggaga cacaaacgtc cccggttgcc tcagcggtgg cggatgccgc 17820ggtgcaggcg gtcgctgcgg ccgcgtccaa gacctctacg gaggtgcaaa cggacccgtg 17880gatgtttcgc gtttcagccc cccggcgccc gcgccgttcg aggaagtacg gcgccgccag 17940cgcgctactg cccgaatatg ccctacatcc ttccattgcg cctacccccg gctatcgtgg 18000ctacacctac cgccccagaa gacgagcaac tacccgacgc cgaaccacca ctggaacccg 18060ccgccgccgt cgccgtcgcc agcccgtgct ggccccgatt tccgtgcgca gggtggctcg 18120cgaaggaggc aggaccctgg tgctgccaac agcgcgctac caccccagca tcgtttaaaa 18180gccggtcttt gtggttcttg cagatatggc cctcacctgc cgcctccgtt tcccggtgcc 18240gggattccga ggaagaatgc accgtaggag gggcatggcc ggccacggcc tgacgggcgg 18300catgcgtcgt gcgcaccacc ggcggcggcg cgcgtcgcac cgtcgcatgc gcggcggtat 18360cctgcccctc cttattccac tgatcgccgc ggcgattggc gccgtgcccg gaattgcatc 18420cgtggccttg caggcgcaga gacactgatt aaaaacaagt tgcatgtgga aaaatcaaaa 18480taaaaagtct ggactctcac gctcgcttgg tcctgtaact attttgtaga atggaagaca 18540tcaactttgc gtctctggcc ccgcgacacg gctcgcgccc gttcatggga aactggcaag 18600atatcggcac cagcaatatg agcggtggcg ccttcagctg gggctcgctg tggagcggca 18660ttaaaaattt cggttccacc gttaagaact atggcagcaa ggcctggaac agcagcacag 18720gccagatgct gagggataag ttgaaagagc aaaatttcca acaaaaggtg gtagatggcc 18780tggcctctgg cattagcggg gtggtggacc tggccaacca ggcagtgcaa aataagatta 18840acagtaagct tgatccccgc cctcccgtag aggagcctcc accggccgtg gagacagtgt 18900ctccagaggg gcgtggcgaa aagcgtccgc gccccgacag ggaagaaact ctggtgacgc 18960aaatagacga gcctccctcg tacgaggagg cactaaagca aggcctgccc accacccgtc 19020ccatcgcgcc catggctacc ggagtgctgg gccagcacac acccgtaacg ctggacctgc 19080ctccccccgc cgacacccag cagaaacctg tgctgccagg cccgaccgcc gttgttgtaa 19140cccgtcctag ccgcgcgtcc ctgcgccgcg ccgccagcgg tccgcgatcg ttgcggcccg 19200tagccagtgg caactggcaa agcacactga acagcatcgt gggtctgggg gtgcaatccc 19260tgaagcgccg acgatgcttc tgatagctaa cgtgtcgtat gtgtgtcatg tatgcgtcca 19320tgtcgccgcc agaggagctg ctgagccgcc gcgcgcccgc tttccaagat ggctacccct 19380tcgatgatgc cgcagtggtc ttacatgcac atctcgggcc aggacgcctc ggagtacctg 19440agccccgggc tggtgcagtt tgcccgcgcc accgagacgt acttcagcct gaataacaag 19500tttagaaacc ccacggtggc gcctacgcac gacgtgacca cagaccggtc ccagcgtttg 19560acgctgcggt tcatccctgt ggaccgtgag gatactgcgt actcgtacaa ggcgcggttc 19620accctagctg tgggtgataa ccgtgtgctg gacatggctt ccacgtactt tgacatccgc 19680ggcgtgctgg acaggggccc tacttttaag ccctactctg gcactgccta caacgccctg 19740gctcccaagg gtgccccaaa tccttgcgaa tgggatgaag ctgctactgc tcttgaaata 19800aacctagaag aagaggacga tgacaacgaa gacgaagtag acgagcaagc tgagcagcaa 19860aaaactcacg tatttgggca ggcgccttat tctggtataa atattacaaa ggagggtatt 19920caaataggtg tcgaaggtca aacacctaaa tatgccgata aaacatttca acctgaacct 19980caaataggag aatctcagtg gtacgaaaca gaaattaatc atgcagctgg gagagtccta 20040aaaaagacta ccccaatgaa accatgttac ggttcatatg caaaacccac aaatgaaaat 20100ggagggcaag gcattcttgt aaagcaacaa aatggaaagc tagaaagtca agtggaaatg 20160caatttttct caactactga ggcagccgca ggcaatggtg ataacttgac tcctaaagtg 20220gtattgtaca gtgaagatgt agatatagaa accccagaca ctcatatttc ttacatgccc 20280actattaagg aaggtaactc acgagaacta atgggccaac aatctatgcc caacaggcct 20340aattacattg cttttaggga caattttatt ggtctaatgt attacaacag cacgggtaat 20400atgggtgttc tggcgggcca agcatcgcag ttgaatgctg ttgtagattt gcaagacaga 20460aacacagagc tttcatacca gcttttgctt gattccattg gtgatagaac caggtacttt 20520tctatgtgga atcaggctgt tgacagctat gatccagatg ttagaattat tgaaaatcat 20580ggaactgaag atgaacttcc aaattactgc tttccactgg gaggtgtgat taatacagag 20640actcttacca aggtaaaacc taaaacaggt caggaaaatg gatgggaaaa agatgctaca 20700gaattttcag ataaaaatga aataagagtt ggaaataatt ttgccatgga aatcaatcta 20760aatgccaacc tgtggagaaa tttcctgtac tccaacatag cgctgtattt gcccgacaag 20820ctaaagtaca gtccttccaa cgtaaaaatt tctgataacc caaacaccta cgactacatg 20880aacaagcgag tggtggctcc cgggctagtg gactgctaca ttaaccttgg agcacgctgg 20940tcccttgact atatggacaa cgtcaaccca tttaaccacc accgcaatgc tggcctgcgc 21000taccgctcaa tgttgctggg caatggtcgc tatgtgccct tccacatcca ggtgcctcag 21060aagttctttg ccattaaaaa cctccttctc ctgccgggct catacaccta cgagtggaac 21120ttcaggaagg atgttaacat ggttctgcag agctccctag gaaatgacct aagggttgac 21180ggagccagca ttaagtttga tagcatttgc ctttacgcca ccttcttccc catggcccac 21240aacaccgcct ccacgcttga ggccatgctt agaaacgaca ccaacgacca gtcctttaac 21300gactatctct ccgccgccaa catgctctac cctatacccg ccaacgctac caacgtgccc 21360atatccatcc cctcccgcaa ctgggcggct ttccgcggct gggccttcac gcgccttaag 21420actaaggaaa ccccatcact gggctcgggc tacgaccctt attacaccta ctctggctct 21480ataccctacc tagatggaac cttttacctc aaccacacct ttaagaaggt ggccattacc 21540tttgactctt ctgtcagctg gcctggcaat gaccgcctgc ttacccccaa cgagtttgaa 21600attaagcgct cagttgacgg ggagggttac aacgttgccc agtgtaacat gaccaaagac 21660tggttcctgg tacaaatgct agctaactat aacattggct accagggctt ctatatccca 21720gagagctaca aggaccgcat gtactccttc tttagaaact tccagcccat gagccgtcag 21780gtggtggatg atactaaata caaggactac caacaggtgg gcatcctaca ccaacacaac 21840aactctggat ttgttggcta ccttgccccc accatgcgcg aaggacaggc ctaccctgct 21900aacttcccct atccgcttat aggcaagacc gcagttgaca gcattaccca gaaaaagttt 21960ctttgcgatc gcaccctttg gcgcatccca ttctccagta actttatgtc catgggcgca 22020ctcacagacc tgggccaaaa ccttctctac gccaactccg cccacgcgct agacatgact 22080tttgaggtgg atcccatgga cgagcccacc cttctttatg ttttgtttga agtctttgac 22140gtggtccgtg tgcaccagcc gcaccgcggc gtcatcgaaa ccgtgtacct gcgcacgccc 22200ttctcggccg gcaacgccac aacataaaga agcaagcaac atcaacaaca gctgccgcca 22260tgggctccag tgagcaggaa ctgaaagcca ttgtcaaaga tcttggttgt gggccatatt 22320ttttgggcac ctatgacaag cgctttccag gctttgtttc tccacacaag ctcgcctgcg 22380ccatagtcaa tacggccggt cgcgagactg ggggcgtaca ctggatggcc tttgcctgga 22440acccgcactc aaaaacatgc tacctctttg agccctttgg cttttctgac cagcgactca 22500agcaggttta ccagtttgag tacgagtcac tcctgcgccg tagcgccatt gcttcttccc 22560ccgaccgctg tataacgctg gaaaagtcca cccaaagcgt acaggggccc aactcggccg 22620cctgtggact attctgctgc

atgtttctcc acgcctttgc caactggccc caaactccca 22680tggatcacaa ccccaccatg aaccttatta ccggggtacc caactccatg ctcaacagtc 22740cccaggtaca gcccaccctg cgtcgcaacc aggaacagct ctacagcttc ctggagcgcc 22800actcgcccta cttccgcagc cacagtgcgc agattaggag cgccacttct ttttgtcact 22860tgaaaaacat gtaaaaataa tgtactagag acactttcaa taaaggcaaa tgcttttatt 22920tgtacactct cgggtgatta tttaccccca cccttgccgt ctgcgccgtt taaaaatcaa 22980aggggttctg ccgcgcatcg ctatgcgcca ctggcaggga cacgttgcga tactggtgtt 23040tagtgctcca cttaaactca ggcacaacca tccgcggcag ctcggtgaag ttttcactcc 23100acaggctgcg caccatcacc aacgcgttta gcaggtcggg cgccgatatc ttgaagtcgc 23160agttggggcc tccgccctgc gcgcgcgagt tgcgatacac agggttgcag cactggaaca 23220ctatcagcgc cgggtggtgc acgctggcca gcacgctctt gtcggagatc agatccgcgt 23280ccaggtcctc cgcgttgctc agggcgaacg gagtcaactt tggtagctgc cttcccaaaa 23340agggcgcgtg cccaggcttt gagttgcact cgcaccgtag tggcatcaaa aggtgaccgt 23400gcccggtctg ggcgttagga tacagcgcct gcataaaagc cttgatctgc ttaaaagcca 23460cctgagcctt tgcgccttca gagaagaaca tgccgcaaga cttgccggaa aactgattgg 23520ccggacaggc cgcgtcgtgc acgcagcacc ttgcgtcggt gttggagatc tgcaccacat 23580ttcggcccca ccggttcttc acgatcttgg ccttgctaga ctgctccttc agcgcgcgct 23640gcccgttttc gctcgtcaca tccatttcaa tcacgtgctc cttatttatc ataatgcttc 23700cgtgtagaca cttaagctcg ccttcgatct cagcgcagcg gtgcagccac aacgcgcagc 23760ccgtgggctc gtgatgcttg taggtcacct ctgcaaacga ctgcaggtac gcctgcagga 23820atcgccccat catcgtcaca aaggtcttgt tgctggtgaa ggtcagctgc aacccgcggt 23880gctcctcgtt cagccaggtc ttgcatacgg ccgccagagc ttccacttgg tcaggcagta 23940gtttgaagtt cgcctttaga tcgttatcca cgtggtactt gtccatcagc gcgcgcgcag 24000cctccatgcc cttctcccac gcagacacga tcggcacact cagcgggttc atcaccgtaa 24060tttcactttc cgcttcgctg ggctcttcct cttcctcttg cgtccgcata ccacgcgcca 24120ctgggtcgtc ttcattcagc cgccgcactg tgcgcttacc tcctttgcca tgcttgatta 24180gcaccggtgg gttgctgaaa cccaccattt gtagcgccac atcttctctt tcttcctcgc 24240tgtccacgat tacctctggt gatggcgggc gctcgggctt gggagaaggg cgcttctttt 24300tcttcttggg cgcaatggcc aaatccgccg ccgaggtcga tggccgcggg ctgggtgtgc 24360gcggcaccag cgcgtcttgt gatgagtctt cctcgtcctc ggactcgata cgccgcctca 24420tccgcttttt tgggggcgcc cggggaggcg gcggcgacgg ggacggggac gacacgtcct 24480ccatggttgg gggacgtcgc gccgcaccgc gtccgcgctc gggggtggtt tcgcgctgct 24540cctcttcccg actggccatt tccttctcct ataggcagaa aaagatcatg gagtcagtcg 24600agaagaagga cagcctaacc gccccctctg agttcgccac caccgcctcc accgatgccg 24660ccaacgcgcc taccaccttc cccgtcgagg cacccccgct tgaggaggag gaagtgatta 24720tcgagcagga cccaggtttt gtaagcgaag acgacgagga ccgctcagta ccaacagagg 24780ataaaaagca agaccaggac aacgcagagg caaacgagga acaagtcggg cggggggacg 24840aaaggcatgg cgactaccta gatgtgggag acgacgtgct gttgaagcat ctgcagcgcc 24900agtgcgccat tatctgcgac gcgttgcaag agcgcagcga tgtgcccctc gccatagcgg 24960atgtcagcct tgcctacgaa cgccacctat tctcaccgcg cgtacccccc aaacgccaag 25020aaaacggcac atgcgagccc aacccgcgcc tcaacttcta ccccgtattt gccgtgccag 25080aggtgcttgc cacctatcac atctttttcc aaaactgcaa gataccccta tcctgccgtg 25140ccaaccgcag ccgagcggac aagcagctgg ccttgcggca gggcgctgtc atacctgata 25200tcgcctcgct caacgaagtg ccaaaaatct ttgagggtct tggacgcgac gagaagcgcg 25260cggcaaacgc tctgcaacag gaaaacagcg aaaatgaaag tcactctgga gtgttggtgg 25320aactcgaggg tgacaacgcg cgcctagccg tactaaaacg cagcatcgag gtcacccact 25380ttgcctaccc ggcacttaac ctacccccca aggtcatgag cacagtcatg agtgagctga 25440tcgtgcgccg tgcgcagccc ctggagaggg atgcaaattt gcaagaacaa acagaggagg 25500gcctacccgc agttggcgac gagcagctag cgcgctggct tcaaacgcgc gagcctgccg 25560acttggagga gcgacgcaaa ctaatgatgg ccgcagtgct cgttaccgtg gagcttgagt 25620gcatgcagcg gttctttgct gacccggaga tgcagcgcaa gctagaggaa acattgcact 25680acacctttcg acagggctac gtacgccagg cctgcaagat ctccaacgtg gagctctgca 25740acctggtctc ctaccttgga attttgcacg aaaaccgcct tgggcaaaac gtgcttcatt 25800ccacgctcaa gggcgaggcg cgccgcgact acgtccgcga ctgcgtttac ttatttctat 25860gctacacctg gcagacggcc atgggcgttt ggcagcagtg cttggaggag tgcaacctca 25920aggagctgca gaaactgcta aagcaaaact tgaaggacct atggacggcc ttcaacgagc 25980gctccgtggc cgcgcacctg gcggacatca ttttccccga acgcctgctt aaaaccctgc 26040aacagggtct gccagacttc accagtcaaa gcatgttgca gaactttagg aactttatcc 26100tagagcgctc aggaatcttg cccgccacct gctgtgcact tcctagcgac tttgtgccca 26160ttaagtaccg cgaatgccct ccgccgcttt ggggccactg ctaccttctg cagctagcca 26220actaccttgc ctaccactct gacataatgg aagacgtgag cggtgacggt ctactggagt 26280gtcactgtcg ctgcaaccta tgcaccccgc accgctccct ggtttgcaat tcgcagctgc 26340ttaacgaaag tcaaattatc ggtacctttg agctgcaggg tccctcgcct gacgaaaagt 26400ccgcggctcc ggggttgaaa ctcactccgg ggctgtggac gtcggcttac cttcgcaaat 26460ttgtacctga ggactaccac gcccacgaga ttaggttcta cgaagaccaa tcccgcccgc 26520ctaatgcgga gcttaccgcc tgcgtcatta cccagggcca cattcttggc caattgcaag 26580ccatcaacaa agcccgccaa gagtttctgc tacgaaaggg acggggggtt tacttggacc 26640cccagtccgg cgaggagctc aacccaatcc ccccgccgcc gcagccctat cagcagcagc 26700cgcgggccct tgcttcccag gatggcaccc aaaaagaagc tgcagctgcc gccgccaccc 26760acggacgagg aggaatactg ggacagtcag gcagaggagg ttttggacga ggaggaggag 26820gacatgatgg aagactggga gagcctagac gaggaagctt ccgaggtcga agaggtgtca 26880gacgaaacac cgtcaccctc ggtcgcattc ccctcgccgg cgccccagaa atcggcaacc 26940ggttccagca tggctacaac ctccgctcct caggcgccgc cggcactgcc cgttcgccga 27000cccaaccgta gatgggacac cactggaacc agggccggta agtccaagca gccgccgccg 27060ttagcccaag agcaacaaca gcgccaaggc taccgctcat ggcgcgggca caagaacgcc 27120atagttgctt gcttgcaaga ctgtgggggc aacatctcct tcgcccgccg ctttcttctc 27180taccatcacg gcgtggcctt cccccgtaac atcctgcatt actaccgtca tctctacagc 27240ccatactgca ccggcggcag cggcagcaac agcagcggcc acacagaagc aaaggcgacc 27300ggatagcaag actctgacaa agcccaagaa atccacagcg gcggcagcag caggaggagg 27360agcgctgcgt ctggcgccca acgaacccgt atcgacccgc gagcttagaa acaggatttt 27420tcccactctg tatgctatat ttcaacagag caggggccaa gaacaagagc tgaaaataaa 27480aaacaggtct ctgcgatccc tcacccgcag ctgcctgtat cacaaaagcg aagatcagct 27540tcggcgcacg ctggaagacg cggaggctct cttcagtaaa tactgcgcgc tgactcttaa 27600ggactagttt cgcgcccttt ctcaaattta agcgcgaaaa ctacgtcatc tccagcggcc 27660acacccggcg ccagcacctg ttgtcagcgc cattatgagc aaggaaattc ccacgcccta 27720catgtggagt taccagccac aaatgggact tgcggctgga gctgcccaag actactcaac 27780ccgaataaac tacatgagcg cgggacccca catgatatcc cgggtcaacg gaatacgcgc 27840ccaccgaaac cgaattctcc tggaacaggc ggctattacc accacacctc gtaataacct 27900taatccccgt agttggcccg ctgccctggt gtaccaggaa agtcccgctc ccaccactgt 27960ggtacttccc agagacgccc aggccgaagt tcagatgact aactcagggg cgcagcttgc 28020gggcggcttt cgtcacaggg tgcggtcgcc cgggcagggt ataactcacc tgacaatcag 28080agggcgaggt attcagctca acgacgagtc ggtgagctcc tcgcttggtc tccgtccgga 28140cgggacattt cagatcggcg gcgccggccg ctcttcattc acgcctcgtc aggcaatcct 28200aactctgcag acctcgtcct ctgagccgcg ctctggaggc attggaactc tgcaatttat 28260tgaggagttt gtgccatcgg tctactttaa ccccttctcg ggacctcccg gccactatcc 28320ggatcaattt attcctaact ttgacgcggt aaaggactcg gcggacggct acgactgaat 28380gttaagtgga gaggcagagc aactgcgcct gaaacacctg gtccactgtc gccgccacaa 28440gtgctttgcc cgcgactccg gtgagttttg ctactttgaa ttgcccgagg atcatatcga 28500gggcccggcg cacggcgtcc ggcttaccgc ccagggagag cttgcccgta gcctgattcg 28560ggagtttacc cagcgccccc tgctagttga gcgggacagg ggaccctgtg ttctcactgt 28620gatttgcaac tgtcctaacc ctggattaca tcaagatctt tgttgccatc tctgtgctga 28680gtataataaa tacagaaatt aaaatatact ggggctccta tcgccatcct gtaaacgcca 28740ccgtcttcac ccgcccaagc aaaccaaggc gaaccttacc tggtactttt aacatctctc 28800cctctgtgat ttacaacagt ttcaacccag acggagtgag tctacgagag aacctctccg 28860agctcagcta ctccatcaga aaaaacacca ccctccttac ctgccgggaa cgtacgagtg 28920cgtcaccggc cgctgcacca cacctaccgc ctgaccgtaa accagacttt ttccggacag 28980acctcaataa ctctgtttac cagaacagga ggtgagctta gaaaaccctt agggtattag 29040gccaaaggcg cagctactgt ggggtttatg aacaattcaa gcaactctac gggctattct 29100aattcaggtt tctctagaat cggggttggg gttattctct gtcttgtgat tctctttatt 29160cttatactaa cgcttctctg cctaaggctc gccgcctgct gtgtgcacat ttgcatttat 29220tgtcagcttt ttaaacgctg gggtcgccac ccaagatgat taggtacata atcctaggtt 29280tactcaccct tgcgtcagcc cacggtacca cccaaaaggt ggattttaag gagccagcct 29340gtaatgttac attcgcagct gaagctaatg agtgcaccac tcttataaaa tgcaccacag 29400aacatgaaaa gctgcttatt cgccacaaaa acaaaattgg caagtatgct gtttatgcta 29460tttggcagcc aggtgacact acagagtata atgttacagt tttccagggt aaaagtcata 29520aaacttttat gtatactttt ccattttatg aaatgtgcga cattaccatg tacatgagca 29580aacagtataa gttgtggccc ccacaaaatt gtgtggaaaa cactggcact ttctgctgca 29640ctgctatgct aattacagtg ctcgctttgg tctgtaccct actctatatt aaatacaaaa 29700gcagacgcag ctttattgag gaaaagaaaa tgccttaatt tactaagtta caaagctaat 29760gtcaccacta actgctttac tcgctgcttg caaaacaaat tcaaaaagtt agcattataa 29820ttagaatagg atttaaaccc cccggtcatt tcctgctcaa taccattccc ctgaacaatt 29880gactctatgt gggatatgct ccagcgctac aaccttgaag tcaggcttcc tggatgtcag 29940catctgactt tggccagcac ctgtcccgcg gatttgttcc agtccaacta cagcgaccca 30000ccctaacaga gatgaccaac acaaccaacg cggccgccgc taccggactt acatctacca 30060caaatacacc ccaagtttct gcctttgtca ataactggga taacttgggc atgtggtggt 30120tctccatagc gcttatgttt gtatgcctta ttattatgtg gctcatctgc tgcctaaagc 30180gcaaacgcgc ccgaccaccc atctatagtc ccatcattgt gctacaccca aacaatgatg 30240gaatccatag attggacgga ctgaaacaca tgttcttttc tcttacagta tgattaaatg 30300agacatgatt cctcgagttt ttatattact gacccttgtt gcgctttttt tgtgcgtgct 30360ccacattggc tgcggtttct cacatcgaag tagactgcat tccagccttc acagtctatt 30420tgctttacgg atttgtcacc ctcacgctca tctgcagcct catcactgtg gtcatcgcct 30480ttatccagtg cattgactgg gtctgtgtgc gctttgcata tctcagacac catccccagt 30540acagggacag gactatagct gagcttctta gaattcttta attatgaaat ttactgtgac 30600ttttctgctg attatttgca ccctatctgc gttttgttcc ccgacctcca agcctcaaag 30660acatatatca tgcagattca ctcgtatatg gaatattcca agttgctaca atgaaaaaag 30720cgatctttcc gaagcctggt tatatgcaat catctctgtt atggtgttct gcagtaccat 30780cttagcccta gctatatatc cctaccttga cattggctgg aacgcaatag atgccatgaa 30840ccacccaact ttccccgcgc ccgctatgct tccactgcaa caagttgttg ccggcggctt 30900tgtcccagcc aatcagcctc gcccaccttc tcccaccccc actgaaatca gctactttaa 30960tctaacagga ggagatgact gacaccctag atctagaaat ggacggaatt attacagagc 31020agcgcctgct agaaagacgc agggcagcgg ccgagcaaca gcgcatgaat caagagctcc 31080aagacatggt taacttgcac cagtgcaaaa ggggtatctt ttgtctggta aagcaggcca 31140aagtcaccta cgacagtaat accaccggac accgccttag ctacaagttg ccaaccaagc 31200gtcagaaatt ggtggtcatg gtgggagaaa agcccattac cataactcag cactcggtag 31260aaaccgaagg ctgcattcac tcaccttgtc aaggacctga ggatctctgc acccttatta 31320agaccctgtg cggtctcaaa gatcttattc cctttaacta ataaaaaaaa ataataaagc 31380atcacttact taaaatcagt tagcaaattt ctgtccagtt tattcagcag cacctccttg 31440ccctcctccc agctctggta ttgcagcttc ctcctggctg caaactttct ccacaatcta 31500aatggaatgt cagtttcctc ctgttcctgt ccatccgcac ccactatctt catgttgttg 31560cagatgaagc gcgcaagacc gtctgaagat accttcaacc ccgtgtatcc atatgacacg 31620gaaaccggtc ctccaactgt gccttttctt actcctccct ttgtatcccc caatgggttt 31680caagagagtc cccctggggt actctctttg cgcctatccg aacctctagt tacctccaat 31740ggcatgcttg cgctcaaaat gggcaacggc ctctctctgg acgaggccgg caaccttacc 31800tcccaaaatg taaccactgt gagcccacct ctcaaaaaaa ccaagtcaaa cataaacctg 31860gaaatatctg cacccctcac agttacctca gaagccctaa ctgtggctgc cgccgcacct 31920ctaatggtcg cgggcaacac actcaccatg caatcacagg ccccgctaac cgtgcacgac 31980tccaaactta gcattgccac ccaaggaccc ctcacagtgt cagaaggaaa gctagccctg 32040caaacatcag gccccctcac caccaccgat agcagtaccc ttactatcac tgcctcaccc 32100cctctaacta ctgccactgg tagcttgggc attgacttga aagagcccat ttatacacaa 32160aatggaaaac taggactaaa gtacggggct cctttgcatg taacagacga cctaaacact 32220ttgaccgtag caactggtcc aggtgtgact attaataata cttccttgca aactaaagtt 32280actggagcct tgggttttga ttcacaaggc aatatgcaac ttaatgtagc aggaggacta 32340aggattgatt ctcaaaacag acgccttata cttgatgtta gttatccgtt tgatgctcaa 32400aaccaactaa atctaagact aggacagggc cctcttttta taaactcagc ccacaacttg 32460gatattaact acaacaaagg cctttacttg tttacagctt caaacaattc caaaaagctt 32520gaggttaacc taagcactgc caaggggttg atgtttgacg ctacagccat agccattaat 32580gcaggagatg ggcttgaatt tggttcacct aatgcaccaa acacaaatcc cctcaaaaca 32640aaaattggcc atggcctaga atttgattca aacaaggcta tggttcctaa actaggaact 32700ggccttagtt ttgacagcac aggtgccatt acagtaggaa acaaaaataa tgataagcta 32760actttgtgga ccacaccagc tccatctcct aactgtagac taaatgcaga gaaagatgct 32820aaactcactt tggtcttaac aaaatgtggc agtcaaatac ttgctacagt ttcagttttg 32880gctgttaaag gcagtttggc tccaatatct ggaacagttc aaagtgctca tcttattata 32940agatttgacg aaaatggagt gctactaaac aattccttcc tggacccaga atattggaac 33000tttagaaatg gagatcttac tgaaggcaca gcctatacaa acgctgttgg atttatgcct 33060aacctatcag cttatccaaa atctcacggt aaaactgcca aaagtaacat tgtcagtcaa 33120gtttacttaa acggagacaa aactaaacct gtaacactaa ccattacact aaacggtaca 33180caggaaacag gagacacaac tccaagtgca tactctatgt cattttcatg ggactggtct 33240ggccacaact acattaatga aatatttgcc acatcctctt acactttttc atacattgcc 33300caagaataaa gaatcgtttg tgttatgttt caacgtgttt atttttcaat tgcagaaaat 33360ttcaagtcat ttttcattca gtagtatagc cccaccacca catagcttat acagatcacc 33420gtaccttaat caaactcaca gaaccctagt attcaacctg ccacctccct cccaacacac 33480agagtacaca gtcctttctc cccggctggc cttaaaaagc atcatatcat gggtaacaga 33540catattctta ggtgttatat tccacacggt ttcctgtcga gccaaacgct catcagtgat 33600attaataaac tccccgggca gctcacttaa gttcatgtcg ctgtccagct gctgagccac 33660aggctgctgt ccaacttgcg gttgcttaac gggcggcgaa ggagaagtcc acgcctacat 33720gggggtagag tcataatcgt gcatcaggat agggcggtgg tgctgcagca gcgcgcgaat 33780aaactgctgc cgccgccgct ccgtcctgca ggaatacaac atggcagtgg tctcctcagc 33840gatgattcgc accgcccgca gcataaggcg ccttgtcctc cgggcacagc agcgcaccct 33900gatctcactt aaatcagcac agtaactgca gcacagcacc acaatattgt tcaaaatccc 33960acagtgcaag gcgctgtatc caaagctcat ggcggggacc acagaaccca cgtggccatc 34020ataccacaag cgcaggtaga ttaagtggcg acccctcata aacacgctgg acataaacat 34080tacctctttt ggcatgttgt aattcaccac ctcccggtac catataaacc tctgattaaa 34140catggcgcca tccaccacca tcctaaacca gctggccaaa acctgcccgc cggctataca 34200ctgcagggaa ccgggactgg aacaatgaca gtggagagcc caggactcgt aaccatggat 34260catcatgctc gtcatgatat caatgttggc acaacacagg cacacgtgca tacacttcct 34320caggattaca agctcctccc gcgttagaac catatcccag ggaacaaccc attcctgaat 34380cagcgtaaat cccacactgc agggaagacc tcgcacgtaa ctcacgttgt gcattgtcaa 34440agtgttacat tcgggcagca gcggatgatc ctccagtatg gtagcgcggg tttctgtctc 34500aaaaggaggt agacgatccc tactgtacgg agtgcgccga gacaaccgag atcgtgttgg 34560tcgtagtgtc atgccaaatg gaacgccgga cgtagtcata tttcctgaag caaaaccagg 34620tgcgggcgtg acaaacagat ctgcgtctcc ggtctcgccg cttagatcgc tctgtgtagt 34680agttgtagta tatccactct ctcaaagcat ccaggcgccc cctggcttcg ggttctatgt 34740aaactccttc atgcgccgct gccctgataa catccaccac cgcagaataa gccacaccca 34800gccaacctac acattcgttc tgcgagtcac acacgggagg agcgggaaga gctggaagaa 34860ccatgttttt ttttttattc caaaagatta tccaaaacct caaaatgaag atctattaag 34920tgaacgcgct cccctccggt ggcgtggtca aactctacag ccaaagaaca gataatggca 34980tttgtaagat gttgcacaat ggcttccaaa aggcaaacgg ccctcacgtc caagtggacg 35040taaaggctaa acccttcagg gtgaatctcc tctataaaca ttccagcacc ttcaaccatg 35100cccaaataat tctcatctcg ccaccttctc aatatatctc taagcaaatc ccgaatatta 35160agtccggcca ttgtaaaaat ctgctccaga gcgccctcca ccttcagcct caagcagcga 35220atcatgattg caaaaattca ggttcctcac agacctgtat aagattcaaa agcggaacat 35280taacaaaaat accgcgatcc cgtaggtccc ttcgcagggc cagctgaaca taatcgtgca 35340ggtctgcacg gaccagcgcg gccacttccc cgccaggaac catgacaaaa gaacccacac 35400tgattatgac acgcatactc ggagctatgc taaccagcgt agccccgatg taagcttgtt 35460gcatgggcgg cgatataaaa tgcaaggtgc tgctcaaaaa atcaggcaaa gcctcgcgca 35520aaaaagaaag cacatcgtag tcatgctcat gcagataaag gcaggtaagc tccggaacca 35580ccacagaaaa agacaccatt tttctctcaa acatgtctgc gggtttctgc ataaacacaa 35640aataaaataa caaaaaaaca tttaaacatt agaagcctgt cttacaacag gaaaaacaac 35700ccttataagc ataagacgga ctacggccat gccggcgtga ccgtaaaaaa actggtcacc 35760gtgattaaaa agcaccaccg acagctcctc ggtcatgtcc ggagtcataa tgtaagactc 35820ggtaaacaca tcaggttgat tcacatcggt cagtgctaaa aagcgaccga aatagcccgg 35880gggaatacat acccgcaggc gtagagacaa cattacagcc cccataggag gtataacaaa 35940attaatagga gagaaaaaca cataaacacc tgaaaaaccc tcctgcctag gcaaaatagc 36000accctcccgc tccagaacaa catacagcgc ttccacagcg gcagccataa cagtcagcct 36060taccagtaaa aaagaaaacc tattaaaaaa acaccactcg acacggcacc agctcaatca 36120gtcacagtgt aaaaaagggc caagtgcaga gcgagtatat ataggactaa aaaatgacgt 36180aacggttaaa gtccacaaaa aacacccaga aaaccgcacg cgaacctacg cccagaaacg 36240aaagccaaaa aacccacaac ttcctcaaat cgtcacttcc gttttcccac gttacgtcac 36300ttcccatttt aagaaaacta caattcccaa cacatacaag ttactccgcc ctaaaaccta 36360cgtcacccgc cccgttccca cgccccgcgc cacgtcacaa actccacccc ctcattatca 36420tattggcttc aatccaaaat aaggtatatt attgatgatg 364601035203DNAArtificial sequencePPE-1-3X-FasC virl construct (lacking repeats) 10catcatcaat aatatacctt attttggatt gaagccaata tgataatgag ggggtggagt 60ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg tagtagtgtg gcggaagtgt 120gatgttgcaa gtgtggcgga acacatgtaa gcgacggatg tggcaaaagt gacgtttttg 180gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg gttttaggcg gatgttgtag 240taaatttggg cgtaaccgag taagatttgg ccattttcgc gggaaaactg aataagagga 300agtgaaatct gaataatttt gtgttactca tagcgcgtaa tatttgtcta gggccgcggg 360gactttgacc gtttacgtgg agactcgccc aggtgttttt ctcaggtgtt ttccgcgttc 420cgggtcaaag ttggcgtttt attattatag tcagtacgta cgtgtacttc tgatcggcga 480tactagggag ataaggatgt acctgacaaa accacattgt tgttgttatc attattattt 540agttttcctt ccttgctaac tcctgacgga atctttctca cctcaaatgc gaagtacttt 600agtttagaaa agacttggtg gaaggggtgg tggtggaaaa gtagggtgat cttccaaact 660aatctggttc cccgcccgcc ccagtagctg ggattcaaga gcgaagagtg gggatcgtcc 720ccttgtttga tcagaaagac ataaaaggaa aatcaagtga acaatgatca gccccacctc 780caccccaccc ccctgcgcgc gcacaataca atctatttaa ttgtacttca tacttttcat 840tccaatgggg tgactttgct tctggagaaa ctcttgattc ttgaactctg gggctggcag 900ctagcctcca gaagcaaagt caccccattg gaatgaaaag tatgaagtac aatgaaaagt 960atgaagtact ggctccagaa gcaaagtcac cctccagaag caaagtcacc ccattggaat 1020gaaaagtatg aagtacgcta gcaaaagggg aagcgggctg ctgctctctg caggttctgc 1080agcggtctct gtctagtggg tgttttcttt ttcttagccc tgcccctgga ttgtcagacg 1140gcgggcgtct

gcctctgaag ttagccgtga tttcctctag agccgggtct tatctctggc 1200tgcacgttgc ctgtgggtga ctaatcacac aataacattg tttagggctg gaataaagtc 1260agagctgttt acccccactc tataggggtt caatataaaa aggcggcgga gaactgtccg 1320agtcagaagc gttcctgcac cggcgctgag agcctgaccc ggtctgctcc gctgtccttg 1380cgcgctgcct cccggctgcc cgcgacgctt tcgccccagt ggaagggcca cttgctgcgg 1440ccgctaattc tgcagatcgg gatccggcat gggcctctcc accgtgcctg acctgctgct 1500gccgctggtg ctcctggagc tgttggtggg aatatacccc tcaggggtta ttggactggt 1560ccctcaccta ggggacaggg agaagagaga tagtgtgtgt ccccaaggaa aatatatcca 1620ccctcaaaat aattcgattt gctgtaccaa gtgccacaaa ggaacctact tgtacaatga 1680ctgtccaggc ccggggcagg atacggactg cagggagtgt gagagcggct ccttcaccgc 1740ttcagaaaac cacctcagac actgcctcag ctgctccaaa tgccgaaagg aaatgggtca 1800ggtggagatc tcttcttgca cagtggaccg ggacaccgtg tgtggctgca ggaagaacca 1860gtaccggcat tattggagtg aaaacctttt ccagtgcttc aattgcagcc tctgcctcaa 1920tgggaccgtg cacctctcct gccaggagaa acagaacacc gtgtgcacct gccatgcagg 1980tttctttcta agagaaaacg agtgtgtctc ctgtagtaac tgtaagaaaa gcctggagtg 2040cacgaagttg tgcctaccaa gcttaggatc cagatctaac ttggggtggc tttgtcttct 2100tcttttgcca attccactaa ttgtttgggt gaagagaaag gaagtacaga aaacatgcag 2160aaagcacaga aaggaaaacc aaggttctca tgaatctcca accttaaatc ctgaaacagt 2220ggcaataaat ttatctgatg ttgacttgag taaatatatc accactattg ctggagtcat 2280gacactaagt caagttaaag gctttgttcg aaagaatggt gtcaatgaag ccaaaataga 2340tgagatcaag aatgacaatg tccaagacac agcagaacag aaagttcaac tgcttcgtaa 2400ttggcatcaa cttcatggaa agaaagaagc gtatgacaca ttgattaaag atctcaaaaa 2460agccaatctt tgtactcttg cagagaaaat tcagactatc atcctcaagg acattactag 2520tgactcagaa aattcaaact tcagaaatga aatccaaagc ttggtctagc tcgagcatgc 2580atctaggcgg ccgcatggca gaaattcgcg aattcgctag cgttaacgga tcctctagac 2640gagatccgaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 2700atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 2760atgtatctta tcatgtctag atctgtactg aaatgtgtgg gcgtggctta agggtgggaa 2820agaatatata aggtgggggt cttatgtagt tttgtatctg ttttgcagca gccgccgccg 2880ccatgagcac caactcgttt gatggaagca ttgtgagctc atatttgaca acgcgcatgc 2940ccccatgggc cggggtgcgt cagaatgtga tgggctccag cattgatggt cgccccgtcc 3000tgcccgcaaa ctctactacc ttgacctacg agaccgtgtc tggaacgccg ttggagactg 3060cagcctccgc cgccgcttca gccgctgcag ccaccgcccg cgggattgtg actgactttg 3120ctttcctgag cccgcttgca agcagtgcag cttcccgttc atccgcccgc gatgacaagt 3180tgacggctct tttggcacaa ttggattctt tgacccggga acttaatgtc gtttctcagc 3240agctgttgga tctgcgccag caggtttctg ccctgaaggc ttcctcccct cccaatgcgg 3300tttaaaacat aaataaaaaa ccagactctg tttggatttg gatcaagcaa gtgtcttgct 3360gtctttattt aggggttttg cgcgcgcggt aggcccggga ccagcggtct cggtcgttga 3420gggtcctgtg tattttttcc aggacgtggt aaaggtgact ctggatgttc agatacatgg 3480gcataagccc gtctctgggg tggaggtagc accactgcag agcttcatgc tgcggggtgg 3540tgttgtagat gatccagtcg tagcaggagc gctgggcgtg gtgcctaaaa atgtctttca 3600gtagcaagct gattgccagg ggcaggccct tggtgtaagt gtttacaaag cggttaagct 3660gggatgggtg catacgtggg gatatgagat gcatcttgga ctgtattttt aggttggcta 3720tgttcccagc catatccctc cggggattca tgttgtgcag aaccaccagc acagtgtatc 3780cggtgcactt gggaaatttg tcatgtagct tagaaggaaa tgcgtggaag aacttggaga 3840cgcccttgtg acctccaaga ttttccatgc attcgtccat aatgatggca atgggcccac 3900gggcggcggc ctgggcgaag atatttctgg gatcactaac gtcatagttg tgttccagga 3960tgagatcgtc ataggccatt tttacaaagc gcgggcggag ggtgccagac tgcggtataa 4020tggttccatc cggcccaggg gcgtagttac cctcacagat ttgcatttcc cacgctttga 4080gttcagatgg ggggatcatg tctacctgcg gggcgatgaa gaaaacggtt tccggggtag 4140gggagatcag ctgggaagaa agcaggttcc tgagcagctg cgacttaccg cagccggtgg 4200gcccgtaaat cacacctatt accggctgca actggtagtt aagagagctg cagctgccgt 4260catccctgag caggggggcc acttcgttaa gcatgtccct gactcgcatg ttttccctga 4320ccaaatccgc cagaaggcgc tcgccgccca gcgatagcag ttcttgcaag gaagcaaagt 4380ttttcaacgg tttgagaccg tccgccgtag gcatgctttt gagcgtttga ccaagcagtt 4440ccaggcggtc ccacagctcg gtcacctgct ctacggcatc tcgatccagc atatctcctc 4500gtttcgcggg ttggggcggc tttcgctgta cggcagtagt cggtgctcgt ccagacgggc 4560cagggtcatg tctttccacg ggcgcagggt cctcgtcagc gtagtctggg tcacggtgaa 4620ggggtgcgct ccgggctgcg cgctggccag ggtgcgcttg aggctggtcc tgctggtgct 4680gaagcgctgc cggtcttcgc cctgcgcgtc ggccaggtag catttgacca tggtgtcata 4740gtccagcccc tccgcggcgt ggcccttggc gcgcagcttg cccttggagg aggcgccgca 4800cgaggggcag tgcagacttt tgagggcgta gagcttgggc gcgagaaata ccgattccgg 4860ggagtaggca tccgcgccgc aggccccgca gacggtctcg cattccacga gccaggtgag 4920ctctggccgt tcggggtcaa aaaccaggtt tcccccatgc tttttgatgc gtttcttacc 4980tctggtttcc atgagccggt gtccacgctc ggtgacgaaa aggctgtccg tgtccccgta 5040tacagacttg agaggcctgt cctcgagcgg tgttccgcgg tcctcctcgt atagaaactc 5100ggaccactct gagacaaagg ctcgcgtcca ggccagcacg aaggaggcta agtgggaggg 5160gtagcggtcg ttgtccacta gggggtccac tcgctccagg gtgtgaagac acatgtcgcc 5220ctcttcggca tcaaggaagg tgattggttt gtaggtgtag gccacgtgac cgggtgttcc 5280tgaagggggg ctataaaagg gggtgggggc gcgttcgtcc tcactctctt ccgcatcgct 5340gtctgcgagg gccagctgtt ggggtgagta ctccctctga aaagcgggca tgacttctgc 5400gctaagattg tcagtttcca aaaacgagga ggatttgata ttcacctggc ccgcggtgat 5460gcctttgagg gtggccgcat ccatctggtc agaaaagaca atctttttgt tgtcaagctt 5520ggtggcaaac gacccgtaga gggcgttgga cagcaacttg gcgatggagc gcagggtttg 5580gtttttgtcg cgatcggcgc gctccttggc cgcgatgttt agctgcacgt attcgcgcgc 5640aacgcaccgc cattcgggaa agacggtggt gcgctcgtcg ggcaccaggt gcacgcgcca 5700accgcggttg tgcagggtga caaggtcaac gctggtggct acctctccgc gtaggcgctc 5760gttggtccag cagaggcggc cgcccttgcg cgagcagaat ggcggtaggg ggtctagctg 5820cgtctcgtcc ggggggtctg cgtccacggt aaagaccccg ggcagcaggc gcgcgtcgaa 5880gtagtctatc ttgcatcctt gcaagtctag cgcctgctgc catgcgcggg cggcaagcgc 5940gcgctcgtat gggttgagtg ggggacccca tggcatgggg tgggtgagcg cggaggcgta 6000catgccgcaa atgtcgtaaa cgtagagggg ctctctgagt attccaagat atgtagggta 6060gcatcttcca ccgcggatgc tggcgcgcac gtaatcgtat agttcgtgcg agggagcgag 6120gaggtcggga ccgaggttgc tacgggcggg ctgctctgct cggaagacta tctgcctgaa 6180gatggcatgt gagttggatg atatggttgg acgctggaag acgttgaagc tggcgtctgt 6240gagacctacc gcgtcacgca cgaaggaggc gtaggagtcg cgcagcttgt tgaccagctc 6300ggcggtgacc tgcacgtcta gggcgcagta gtccagggtt tccttgatga tgtcatactt 6360atcctgtccc ttttttttcc acagctcgcg gttgaggaca aactcttcgc ggtctttcca 6420gtactcttgg atcggaaacc cgtcggcctc cgaacggtaa gagcctagca tgtagaactg 6480gttgacggcc tggtaggcgc agcatccctt ttctacgggt agcgcgtatg cctgcgcggc 6540cttccggagc gaggtgtggg tgagcgcaaa ggtgtccctg accatgactt tgaggtactg 6600gtatttgaag tcagtgtcgt cgcatccgcc ctgctcccag agcaaaaagt ccgtgcgctt 6660tttggaacgc ggatttggca gggcgaaggt gacatcgttg aagagtatct ttcccgcgcg 6720aggcataaag ttgcgtgtga tgcggaaggg tcccggcacc tcggaacggt tgttaattac 6780ctgggcggcg agcacgatct cgtcaaagcc gttgatgttg tggcccacaa tgtaaagttc 6840caagaagcgc gggatgccct tgatggaagg caatttttta agttcctcgt aggtgagctc 6900ttcaggggag ctgagcccgt gctctgaaag ggcccagtct gcaagatgag ggttggaagc 6960gacgaatgag ctccacaggt cacgggccat tagcatttgc aggtggtcgc gaaaggtcct 7020aaactggcga cctatggcca ttttttctgg ggtgatgcag tagaaggtaa gcgggtcttg 7080ttcccagcgg tcccatccaa ggttcgcggc taggtctcgc gcggcagtca ctagaggctc 7140atctccgccg aacttcatga ccagcatgaa gggcacgagc tgcttcccaa aggcccccat 7200ccaagtatag gtctctacat cgtaggtgac aaagagacgc tcggtgcgag gatgcgagcc 7260gatcgggaag aactggatct cccgccacca attggaggag tggctattga tgtggtgaaa 7320gtagaagtcc ctgcgacggg ccgaacactc gtgctggctt ttgtaaaaac gtgcgcagta 7380ctggcagcgg tgcacgggct gtacatcctg cacgaggttg acctgacgac cgcgcacaag 7440gaagcagagt gggaatttga gcccctcgcc tggcgggttt ggctggtggt cttctacttc 7500ggctgcttgt ccttgaccgt ctggctgctc gaggggagtt acggtggatc ggaccaccac 7560gccgcgcgag cccaaagtcc agatgtccgc gcgcggcggt cggagcttga tgacaacatc 7620gcgcagatgg gagctgtcca tggtctggag ctcccgcggc gtcaggtcag gcgggagctc 7680ctgcaggttt acctcgcata gacgggtcag ggcgcgggct agatccaggt gatacctaat 7740ttccaggggc tggttggtgg cggcgtcgat ggcttgcaag aggccgcatc cccgcggcgc 7800gactacggta ccgcgcggcg ggcggtgggc cgcgggggtg tccttggatg atgcatctaa 7860aagcggtgac gcgggcgagc ccccggaggt agggggggct ccggacccgc cgggagaggg 7920ggcaggggca cgtcggcgcc gcgcgcgggc aggagctggt gctgcgcgcg taggttgctg 7980gcgaacgcga cgacgcggcg gttgatctcc tgaatctggc gcctctgcgt gaagacgacg 8040ggcccggtga gcttgaacct gaaagagagt tcgacagaat caatttcggt gtcgttgacg 8100gcggcctggc gcaaaatctc ctgcacgtct cctgagttgt cttgataggc gatctcggcc 8160atgaactgct cgatctcttc ctcctggaga tctccgcgtc cggctcgctc cacggtggcg 8220gcgaggtcgt tggaaatgcg ggccatgagc tgcgagaagg cgttgaggcc tccctcgttc 8280cagacgcggc tgtagaccac gcccccttcg gcatcgcggg cgcgcatgac cacctgcgcg 8340agattgagct ccacgtgccg ggcgaagacg gcgtagtttc gcaggcgctg aaagaggtag 8400ttgagggtgg tggcggtgtg ttctgccacg aagaagtaca taacccagcg tcgcaacgtg 8460gattcgttga tatcccccaa ggcctcaagg cgctccatgg cctcgtagaa gtccacggcg 8520aagttgaaaa actgggagtt gcgcgccgac acggttaact cctcctccag aagacggatg 8580agctcggcga cagtgtcgcg cacctcgcgc tcaaaggcta caggggcctc ttcttcttct 8640tcaatctcct cttccataag ggcctcccct tcttcttctt ctggcggcgg tgggggaggg 8700gggacacggc ggcgacgacg gcgcaccggg aggcggtcga caaagcgctc gatcatctcc 8760ccgcggcgac ggcgcatggt ctcggtgacg gcgcggccgt tctcgcgggg gcgcagttgg 8820aagacgccgc ccgtcatgtc ccggttatgg gttggcgggg ggctgccatg cggcagggat 8880acggcgctaa cgatgcatct caacaattgt tgtgtaggta ctccgccgcc gagggacctg 8940agcgagtccg catcgaccgg atcggaaaac ctctcgagaa aggcgtctaa ccagtcacag 9000tcgcaaggta ggctgagcac cgtggcgggc ggcagcgggc ggcggtcggg gttgtttctg 9060gcggaggtgc tgctgatgat gtaattaaag taggcggtct tgagacggcg gatggtcgac 9120agaagcacca tgtccttggg tccggcctgc tgaatgcgca ggcggtcggc catgccccag 9180gcttcgtttt gacatcggcg caggtctttg tagtagtctt gcatgagcct ttctaccggc 9240acttcttctt ctccttcctc ttgtcctgca tctcttgcat ctatcgctgc ggcggcggcg 9300gagtttggcc gtaggtggcg ccctcttcct cccatgcgtg tgaccccgaa gcccctcatc 9360ggctgaagca gggctaggtc ggcgacaacg cgctcggcta atatggcctg ctgcacctgc 9420gtgagggtag actggaagtc atccatgtcc acaaagcggt ggtatgcgcc cgtgttgatg 9480gtgtaagtgc agttggccat aacggaccag ttaacggtct ggtgacccgg ctgcgagagc 9540tcggtgtacc tgagacgcga gtaagccctc gagtcaaata cgtagtcgtt gcaagtccgc 9600accaggtact ggtatcccac caaaaagtgc ggcggcggct ggcggtagag gggccagcgt 9660agggtggccg gggctccggg ggcgagatct tccaacataa ggcgatgata tccgtagatg 9720tacctggaca tccaggtgat gccggcggcg gtggtggagg cgcgcggaaa gtcgcggacg 9780cggttccaga tgttgcgcag cggcaaaaag tgctccatgg tcgggacgct ctggccggtc 9840aggcgcgcgc aatcgttgac gctctagacc gtgcaaaagg agagcctgta agcgggcact 9900cttccgtggt ctggtggata aattcgcaag ggtatcatgg cggacgaccg gggttcgagc 9960cccgtatccg gccgtccgcc gtgatccatg cggttaccgc ccgcgtgtcg aacccaggtg 10020tgcgacgtca gacaacgggg gagtgctcct tttggcttcc ttccaggcgc ggcggctgct 10080gcgctagctt ttttggccac tggccgcgcg cagcgtaagc ggttaggctg gaaagcgaaa 10140gcattaagtg gctcgctccc tgtagccgga gggttatttt ccaagggttg agtcgcggga 10200cccccggttc gagtctcgga ccggccggac tgcggcgaac gggggtttgc ctccccgtca 10260tgcaagaccc cgcttgcaaa ttcctccgga aacagggacg agcccctttt ttgcttttcc 10320cagatgcatc cggtgctgcg gcagatgcgc ccccctcctc agcagcggca agagcaagag 10380cagcggcaga catgcagggc accctcccct cctcctaccg cgtcaggagg ggcgacatcc 10440gcggttgacg cggcagcaga tggtgattac gaacccccgc ggcgccgggc ccggcactac 10500ctggacttgg aggagggcga gggcctggcg cggctaggag cgccctctcc tgagcggcac 10560ccaagggtgc agctgaagcg tgatacgcgt gaggcgtacg tgccgcggca gaacctgttt 10620cgcgaccgcg agggagagga gcccgaggag atgcgggatc gaaagttcca cgcagggcgc 10680gagctgcggc atggcctgaa tcgcgagcgg ttgctgcgcg aggaggactt tgagcccgac 10740gcgcgaaccg ggattagtcc cgcgcgcgca cacgtggcgg ccgccgacct ggtaaccgca 10800tacgagcaga cggtgaacca ggagattaac tttcaaaaaa gctttaacaa ccacgtgcgt 10860acgcttgtgg cgcgcgagga ggtggctata ggactgatgc atctgtggga ctttgtaagc 10920gcgctggagc aaaacccaaa tagcaagccg ctcatggcgc agctgttcct tatagtgcag 10980cacagcaggg acaacgaggc attcagggat gcgctgctaa acatagtaga gcccgagggc 11040cgctggctgc tcgatttgat aaacatcctg cagagcatag tggtgcagga gcgcagcttg 11100agcctggctg acaaggtggc cgccatcaac tattccatgc ttagcctggg caagttttac 11160gcccgcaaga tataccatac cccttacgtt cccatagaca aggaggtaaa gatcgagggg 11220ttctacatgc gcatggcgct gaaggtgctt accttgagcg acgacctggg cgtttatcgc 11280aacgagcgca tccacaaggc cgtgagcgtg agccggcggc gcgagctcag cgaccgcgag 11340ctgatgcaca gcctgcaaag ggccctggct ggcacgggca gcggcgatag agaggccgag 11400tcctactttg acgcgggcgc tgacctgcgc tgggccccaa gccgacgcgc cctggaggca 11460gctggggccg gacctgggct ggcggtggca cccgcgcgcg ctggcaacgt cggcggcgtg 11520gaggaatatg acgaggacga tgagtacgag ccagaggacg gcgagtacta agcggtgatg 11580tttctgatca gatgatgcaa gacgcaacgg acccggcggt gcgggcggcg ctgcagagcc 11640agccgtccgg ccttaactcc acggacgact ggcgccaggt catggaccgc atcatgtcgc 11700tgactgcgcg caatcctgac gcgttccggc agcagccgca ggccaaccgg ctctccgcaa 11760ttctggaagc ggtggtcccg gcgcgcgcaa accccacgca cgagaaggtg ctggcgatcg 11820taaacgcgct ggccgaaaac agggccatcc ggcccgacga ggccggcctg gtctacgacg 11880cgctgcttca gcgcgtggct cgttacaaca gcggcaacgt gcagaccaac ctggaccggc 11940tggtggggga tgtgcgcgag gccgtggcgc agcgtgagcg cgcgcagcag cagggcaacc 12000tgggctccat ggttgcacta aacgccttcc tgagtacaca gcccgccaac gtgccgcggg 12060gacaggagga ctacaccaac tttgtgagcg cactgcggct aatggtgact gagacaccgc 12120aaagtgaggt gtaccagtct gggccagact attttttcca gaccagtaga caaggcctgc 12180agaccgtaaa cctgagccag gctttcaaaa acttgcaggg gctgtggggg gtgcgggctc 12240ccacaggcga ccgcgcgacc gtgtctagct tgctgacgcc caactcgcgc ctgttgctgc 12300tgctaatagc gcccttcacg gacagtggca gcgtgtcccg ggacacatac ctaggtcact 12360tgctgacact gtaccgcgag gccataggtc aggcgcatgt ggacgagcat actttccagg 12420agattacaag tgtcagccgc gcgctggggc aggaggacac gggcagcctg gaggcaaccc 12480taaactacct gctgaccaac cggcggcaga agatcccctc gttgcacagt ttaaacagcg 12540aggaggagcg cattttgcgc tacgtgcagc agagcgtgag ccttaacctg atgcgcgacg 12600gggtaacgcc cagcgtggcg ctggacatga ccgcgcgcaa catggaaccg ggcatgtatg 12660cctcaaaccg gccgtttatc aaccgcctaa tggactactt gcatcgcgcg gccgccgtga 12720accccgagta tttcaccaat gccatcttga acccgcactg gctaccgccc cctggtttct 12780acaccggggg attcgaggtg cccgagggta acgatggatt cctctgggac gacatagacg 12840acagcgtgtt ttccccgcaa ccgcagaccc tgctagagtt gcaacagcgc gagcaggcag 12900aggcggcgct gcgaaaggaa agcttccgca ggccaagcag cttgtccgat ctaggcgctg 12960cggccccgcg gtcagatgct agtagcccat ttccaagctt gatagggtct cttaccagca 13020ctcgcaccac ccgcccgcgc ctgctgggcg aggaggagta cctaaacaac tcgctgctgc 13080agccgcagcg cgaaaaaaac ctgcctccgg catttcccaa caacgggata gagagcctag 13140tggacaagat gagtagatgg aagacgtacg cgcaggagca cagggacgtg ccaggcccgc 13200gcccgcccac ccgtcgtcaa aggcacgacc gtcagcgggg tctggtgtgg gaggacgatg 13260actcggcaga cgacagcagc gtcctggatt tgggagggag tggcaacccg tttgcgcacc 13320ttcgccccag gctggggaga atgttttaaa aaaaaaaaaa gcatgatgca aaataaaaaa 13380ctcaccaagg ccatggcacc gagcgttggt tttcttgtat tccccttagt atgcggcgcg 13440cggcgatgta tgaggaaggt cctcctccct cctacgagag tgtggtgagc gcggcgccag 13500tggcggcggc gctgggttct cccttcgatg ctcccctgga cccgccgttt gtgcctccgc 13560ggtacctgcg gcctaccggg gggagaaaca gcatccgtta ctctgagttg gcacccctat 13620tcgacaccac ccgtgtgtac ctggtggaca acaagtcaac ggatgtggca tccctgaact 13680accagaacga ccacagcaac tttctgacca cggtcattca aaacaatgac tacagcccgg 13740gggaggcaag cacacagacc atcaatcttg acgaccggtc gcactggggc ggcgacctga 13800aaaccatcct gcataccaac atgccaaatg tgaacgagtt catgtttacc aataagttta 13860aggcgcgggt gatggtgtcg cgcttgccta ctaaggacaa tcaggtggag ctgaaatacg 13920agtgggtgga gttcacgctg cccgagggca actactccga gaccatgacc atagacctta 13980tgaacaacgc gatcgtggag cactacttga aagtgggcag acagaacggg gttctggaaa 14040gcgacatcgg ggtaaagttt gacacccgca acttcagact ggggtttgac cccgtcactg 14100gtcttgtcat gcctggggta tatacaaacg aagccttcca tccagacatc attttgctgc 14160caggatgcgg ggtggacttc acccacagcc gcctgagcaa cttgttgggc atccgcaagc 14220ggcaaccctt ccaggagggc tttaggatca cctacgatga tctggagggt ggtaacattc 14280ccgcactgtt ggatgtggac gcctaccagg cgagcttgaa agatgacacc gaacagggcg 14340ggggtggcgc aggcggcagc aacagcagtg gcagcggcgc ggaagagaac tccaacgcgg 14400cagccgcggc aatgcagccg gtggaggaca tgaacgatca tgccattcgc ggcgacacct 14460ttgccacacg ggctgaggag aagcgcgctg aggccgaagc agcggccgaa gctgccgccc 14520ccgctgcgca acccgaggtc gagaagcctc agaagaaacc ggtgatcaaa cccctgacag 14580aggacagcaa gaaacgcagt tacaacctaa taagcaatga cagcaccttc acccagtacc 14640gcagctggta ccttgcatac aactacggcg accctcagac cggaatccgc tcatggaccc 14700tgctttgcac tcctgacgta acctgcggct cggagcaggt ctactggtcg ttgccagaca 14760tgatgcaaga ccccgtgacc ttccgctcca cgcgccagat cagcaacttt ccggtggtgg 14820gcgccgagct gttgcccgtg cactccaaga gcttctacaa cgaccaggcc gtctactccc 14880aactcatccg ccagtttacc tctctgaccc acgtgttcaa tcgctttccc gagaaccaga 14940ttttggcgcg cccgccagcc cccaccatca ccaccgtcag tgaaaacgtt cctgctctca 15000cagatcacgg gacgctaccg ctgcgcaaca gcatcggagg agtccagcga gtgaccatta 15060ctgacgccag acgccgcacc tgcccctacg tttacaaggc cctgggcata gtctcgccgc 15120gcgtcctatc gagccgcact ttttgagcaa gcatgtccat ccttatatcg cccagcaata 15180acacaggctg gggcctgcgc ttcccaagca agatgtttgg cggggccaag aagcgctccg 15240accaacaccc agtgcgcgtg cgcgggcact accgcgcgcc ctggggcgcg cacaaacgcg 15300gccgcactgg gcgcaccacc gtcgatgacg ccatcgacgc ggtggtggag gaggcgcgca 15360actacacgcc cacgccgcca ccagtgtcca cagtggacgc ggccattcag accgtggtgc 15420gcggagcccg gcgctatgct aaaatgaaga gacggcggag gcgcgtagca cgtcgccacc 15480gccgccgacc cggcactgcc gcccaacgcg cggcggcggc cctgcttaac cgcgcacgtc 15540gcaccggccg acgggcggcc atgcgggccg ctcgaaggct ggccgcgggt attgtcactg 15600tgccccccag gtccaggcga cgagcggccg ccgcagcagc cgcggccatt agtgctatga 15660ctcagggtcg caggggcaac gtgtattggg tgcgcgactc ggttagcggc ctgcgcgtgc 15720ccgtgcgcac ccgccccccg cgcaactaga ttgcaagaaa aaactactta gactcgtact 15780gttgtatgta tccagcggcg gcggcgcgca acgaagctat gtccaagcgc aaaatcaaag 15840aagagatgct ccaggtcatc gcgccggaga tctatggccc cccgaagaag gaagagcagg 15900attacaagcc ccgaaagcta aagcgggtca aaaagaaaaa gaaagatgat gatgatgaac 15960ttgacgacga ggtggaactg ctgcacgcta ccgcgcccag gcgacgggta cagtggaaag 16020gtcgacgcgt aaaacgtgtt ttgcgacccg gcaccaccgt agtctttacg cccggtgagc 16080gctccacccg cacctacaag cgcgtgtatg atgaggtgta cggcgacgag gacctgcttg 16140agcaggccaa cgagcgcctc ggggagtttg cctacggaaa gcggcataag gacatgctgg 16200cgttgccgct

ggacgagggc aacccaacac ctagcctaaa gcccgtaaca ctgcagcagg 16260tgctgcccgc gcttgcaccg tccgaagaaa agcgcggcct aaagcgcgag tctggtgact 16320tggcacccac cgtgcagctg atggtaccca agcgccagcg actggaagat gtcttggaaa 16380aaatgaccgt ggaacctggg ctggagcccg aggtccgcgt gcggccaatc aagcaggtgg 16440cgccgggact gggcgtgcag accgtggacg ttcagatacc cactaccagt agcaccagta 16500ttgccaccgc cacagagggc atggagacac aaacgtcccc ggttgcctca gcggtggcgg 16560atgccgcggt gcaggcggtc gctgcggccg cgtccaagac ctctacggag gtgcaaacgg 16620acccgtggat gtttcgcgtt tcagcccccc ggcgcccgcg ccgttcgagg aagtacggcg 16680ccgccagcgc gctactgccc gaatatgccc tacatccttc cattgcgcct acccccggct 16740atcgtggcta cacctaccgc cccagaagac gagcaactac ccgacgccga accaccactg 16800gaacccgccg ccgccgtcgc cgtcgccagc ccgtgctggc cccgatttcc gtgcgcaggg 16860tggctcgcga aggaggcagg accctggtgc tgccaacagc gcgctaccac cccagcatcg 16920tttaaaagcc ggtctttgtg gttcttgcag atatggccct cacctgccgc ctccgtttcc 16980cggtgccggg attccgagga agaatgcacc gtaggagggg catggccggc cacggcctga 17040cgggcggcat gcgtcgtgcg caccaccggc ggcggcgcgc gtcgcaccgt cgcatgcgcg 17100gcggtatcct gcccctcctt attccactga tcgccgcggc gattggcgcc gtgcccggaa 17160ttgcatccgt ggccttgcag gcgcagagac actgattaaa aacaagttgc atgtggaaaa 17220atcaaaataa aaagtctgga ctctcacgct cgcttggtcc tgtaactatt ttgtagaatg 17280gaagacatca actttgcgtc tctggccccg cgacacggct cgcgcccgtt catgggaaac 17340tggcaagata tcggcaccag caatatgagc ggtggcgcct tcagctgggg ctcgctgtgg 17400agcggcatta aaaatttcgg ttccaccgtt aagaactatg gcagcaaggc ctggaacagc 17460agcacaggcc agatgctgag ggataagttg aaagagcaaa atttccaaca aaaggtggta 17520gatggcctgg cctctggcat tagcggggtg gtggacctgg ccaaccaggc agtgcaaaat 17580aagattaaca gtaagcttga tccccgccct cccgtagagg agcctccacc ggccgtggag 17640acagtgtctc cagaggggcg tggcgaaaag cgtccgcgcc ccgacaggga agaaactctg 17700gtgacgcaaa tagacgagcc tccctcgtac gaggaggcac taaagcaagg cctgcccacc 17760acccgtccca tcgcgcccat ggctaccgga gtgctgggcc agcacacacc cgtaacgctg 17820gacctgcctc cccccgccga cacccagcag aaacctgtgc tgccaggccc gaccgccgtt 17880gttgtaaccc gtcctagccg cgcgtccctg cgccgcgccg ccagcggtcc gcgatcgttg 17940cggcccgtag ccagtggcaa ctggcaaagc acactgaaca gcatcgtggg tctgggggtg 18000caatccctga agcgccgacg atgcttctga tagctaacgt gtcgtatgtg tgtcatgtat 18060gcgtccatgt cgccgccaga ggagctgctg agccgccgcg cgcccgcttt ccaagatggc 18120taccccttcg atgatgccgc agtggtctta catgcacatc tcgggccagg acgcctcgga 18180gtacctgagc cccgggctgg tgcagtttgc ccgcgccacc gagacgtact tcagcctgaa 18240taacaagttt agaaacccca cggtggcgcc tacgcacgac gtgaccacag accggtccca 18300gcgtttgacg ctgcggttca tccctgtgga ccgtgaggat actgcgtact cgtacaaggc 18360gcggttcacc ctagctgtgg gtgataaccg tgtgctggac atggcttcca cgtactttga 18420catccgcggc gtgctggaca ggggccctac ttttaagccc tactctggca ctgcctacaa 18480cgccctggct cccaagggtg ccccaaatcc ttgcgaatgg gatgaagctg ctactgctct 18540tgaaataaac ctagaagaag aggacgatga caacgaagac gaagtagacg agcaagctga 18600gcagcaaaaa actcacgtat ttgggcaggc gccttattct ggtataaata ttacaaagga 18660gggtattcaa ataggtgtcg aaggtcaaac acctaaatat gccgataaaa catttcaacc 18720tgaacctcaa ataggagaat ctcagtggta cgaaacagaa attaatcatg cagctgggag 18780agtcctaaaa aagactaccc caatgaaacc atgttacggt tcatatgcaa aacccacaaa 18840tgaaaatgga gggcaaggca ttcttgtaaa gcaacaaaat ggaaagctag aaagtcaagt 18900ggaaatgcaa tttttctcaa ctactgaggc agccgcaggc aatggtgata acttgactcc 18960taaagtggta ttgtacagtg aagatgtaga tatagaaacc ccagacactc atatttctta 19020catgcccact attaaggaag gtaactcacg agaactaatg ggccaacaat ctatgcccaa 19080caggcctaat tacattgctt ttagggacaa ttttattggt ctaatgtatt acaacagcac 19140gggtaatatg ggtgttctgg cgggccaagc atcgcagttg aatgctgttg tagatttgca 19200agacagaaac acagagcttt cataccagct tttgcttgat tccattggtg atagaaccag 19260gtacttttct atgtggaatc aggctgttga cagctatgat ccagatgtta gaattattga 19320aaatcatgga actgaagatg aacttccaaa ttactgcttt ccactgggag gtgtgattaa 19380tacagagact cttaccaagg taaaacctaa aacaggtcag gaaaatggat gggaaaaaga 19440tgctacagaa ttttcagata aaaatgaaat aagagttgga aataattttg ccatggaaat 19500caatctaaat gccaacctgt ggagaaattt cctgtactcc aacatagcgc tgtatttgcc 19560cgacaagcta aagtacagtc cttccaacgt aaaaatttct gataacccaa acacctacga 19620ctacatgaac aagcgagtgg tggctcccgg gctagtggac tgctacatta accttggagc 19680acgctggtcc cttgactata tggacaacgt caacccattt aaccaccacc gcaatgctgg 19740cctgcgctac cgctcaatgt tgctgggcaa tggtcgctat gtgcccttcc acatccaggt 19800gcctcagaag ttctttgcca ttaaaaacct ccttctcctg ccgggctcat acacctacga 19860gtggaacttc aggaaggatg ttaacatggt tctgcagagc tccctaggaa atgacctaag 19920ggttgacgga gccagcatta agtttgatag catttgcctt tacgccacct tcttccccat 19980ggcccacaac accgcctcca cgcttgaggc catgcttaga aacgacacca acgaccagtc 20040ctttaacgac tatctctccg ccgccaacat gctctaccct atacccgcca acgctaccaa 20100cgtgcccata tccatcccct cccgcaactg ggcggctttc cgcggctggg ccttcacgcg 20160ccttaagact aaggaaaccc catcactggg ctcgggctac gacccttatt acacctactc 20220tggctctata ccctacctag atggaacctt ttacctcaac cacaccttta agaaggtggc 20280cattaccttt gactcttctg tcagctggcc tggcaatgac cgcctgctta cccccaacga 20340gtttgaaatt aagcgctcag ttgacgggga gggttacaac gttgcccagt gtaacatgac 20400caaagactgg ttcctggtac aaatgctagc taactataac attggctacc agggcttcta 20460tatcccagag agctacaagg accgcatgta ctccttcttt agaaacttcc agcccatgag 20520ccgtcaggtg gtggatgata ctaaatacaa ggactaccaa caggtgggca tcctacacca 20580acacaacaac tctggatttg ttggctacct tgcccccacc atgcgcgaag gacaggccta 20640ccctgctaac ttcccctatc cgcttatagg caagaccgca gttgacagca ttacccagaa 20700aaagtttctt tgcgatcgca ccctttggcg catcccattc tccagtaact ttatgtccat 20760gggcgcactc acagacctgg gccaaaacct tctctacgcc aactccgccc acgcgctaga 20820catgactttt gaggtggatc ccatggacga gcccaccctt ctttatgttt tgtttgaagt 20880ctttgacgtg gtccgtgtgc accagccgca ccgcggcgtc atcgaaaccg tgtacctgcg 20940cacgcccttc tcggccggca acgccacaac ataaagaagc aagcaacatc aacaacagct 21000gccgccatgg gctccagtga gcaggaactg aaagccattg tcaaagatct tggttgtggg 21060ccatattttt tgggcaccta tgacaagcgc tttccaggct ttgtttctcc acacaagctc 21120gcctgcgcca tagtcaatac ggccggtcgc gagactgggg gcgtacactg gatggccttt 21180gcctggaacc cgcactcaaa aacatgctac ctctttgagc cctttggctt ttctgaccag 21240cgactcaagc aggtttacca gtttgagtac gagtcactcc tgcgccgtag cgccattgct 21300tcttcccccg accgctgtat aacgctggaa aagtccaccc aaagcgtaca ggggcccaac 21360tcggccgcct gtggactatt ctgctgcatg tttctccacg cctttgccaa ctggccccaa 21420actcccatgg atcacaaccc caccatgaac cttattaccg gggtacccaa ctccatgctc 21480aacagtcccc aggtacagcc caccctgcgt cgcaaccagg aacagctcta cagcttcctg 21540gagcgccact cgccctactt ccgcagccac agtgcgcaga ttaggagcgc cacttctttt 21600tgtcacttga aaaacatgta aaaataatgt actagagaca ctttcaataa aggcaaatgc 21660ttttatttgt acactctcgg gtgattattt acccccaccc ttgccgtctg cgccgtttaa 21720aaatcaaagg ggttctgccg cgcatcgcta tgcgccactg gcagggacac gttgcgatac 21780tggtgtttag tgctccactt aaactcaggc acaaccatcc gcggcagctc ggtgaagttt 21840tcactccaca ggctgcgcac catcaccaac gcgtttagca ggtcgggcgc cgatatcttg 21900aagtcgcagt tggggcctcc gccctgcgcg cgcgagttgc gatacacagg gttgcagcac 21960tggaacacta tcagcgccgg gtggtgcacg ctggccagca cgctcttgtc ggagatcaga 22020tccgcgtcca ggtcctccgc gttgctcagg gcgaacggag tcaactttgg tagctgcctt 22080cccaaaaagg gcgcgtgccc aggctttgag ttgcactcgc accgtagtgg catcaaaagg 22140tgaccgtgcc cggtctgggc gttaggatac agcgcctgca taaaagcctt gatctgctta 22200aaagccacct gagcctttgc gccttcagag aagaacatgc cgcaagactt gccggaaaac 22260tgattggccg gacaggccgc gtcgtgcacg cagcaccttg cgtcggtgtt ggagatctgc 22320accacatttc ggccccaccg gttcttcacg atcttggcct tgctagactg ctccttcagc 22380gcgcgctgcc cgttttcgct cgtcacatcc atttcaatca cgtgctcctt atttatcata 22440atgcttccgt gtagacactt aagctcgcct tcgatctcag cgcagcggtg cagccacaac 22500gcgcagcccg tgggctcgtg atgcttgtag gtcacctctg caaacgactg caggtacgcc 22560tgcaggaatc gccccatcat cgtcacaaag gtcttgttgc tggtgaaggt cagctgcaac 22620ccgcggtgct cctcgttcag ccaggtcttg catacggccg ccagagcttc cacttggtca 22680ggcagtagtt tgaagttcgc ctttagatcg ttatccacgt ggtacttgtc catcagcgcg 22740cgcgcagcct ccatgccctt ctcccacgca gacacgatcg gcacactcag cgggttcatc 22800accgtaattt cactttccgc ttcgctgggc tcttcctctt cctcttgcgt ccgcatacca 22860cgcgccactg ggtcgtcttc attcagccgc cgcactgtgc gcttacctcc tttgccatgc 22920ttgattagca ccggtgggtt gctgaaaccc accatttgta gcgccacatc ttctctttct 22980tcctcgctgt ccacgattac ctctggtgat ggcgggcgct cgggcttggg agaagggcgc 23040ttctttttct tcttgggcgc aatggccaaa tccgccgccg aggtcgatgg ccgcgggctg 23100ggtgtgcgcg gcaccagcgc gtcttgtgat gagtcttcct cgtcctcgga ctcgatacgc 23160cgcctcatcc gcttttttgg gggcgcccgg ggaggcggcg gcgacgggga cggggacgac 23220acgtcctcca tggttggggg acgtcgcgcc gcaccgcgtc cgcgctcggg ggtggtttcg 23280cgctgctcct cttcccgact ggccatttcc ttctcctata ggcagaaaaa gatcatggag 23340tcagtcgaga agaaggacag cctaaccgcc ccctctgagt tcgccaccac cgcctccacc 23400gatgccgcca acgcgcctac caccttcccc gtcgaggcac ccccgcttga ggaggaggaa 23460gtgattatcg agcaggaccc aggttttgta agcgaagacg acgaggaccg ctcagtacca 23520acagaggata aaaagcaaga ccaggacaac gcagaggcaa acgaggaaca agtcgggcgg 23580ggggacgaaa ggcatggcga ctacctagat gtgggagacg acgtgctgtt gaagcatctg 23640cagcgccagt gcgccattat ctgcgacgcg ttgcaagagc gcagcgatgt gcccctcgcc 23700atagcggatg tcagccttgc ctacgaacgc cacctattct caccgcgcgt accccccaaa 23760cgccaagaaa acggcacatg cgagcccaac ccgcgcctca acttctaccc cgtatttgcc 23820gtgccagagg tgcttgccac ctatcacatc tttttccaaa actgcaagat acccctatcc 23880tgccgtgcca accgcagccg agcggacaag cagctggcct tgcggcaggg cgctgtcata 23940cctgatatcg cctcgctcaa cgaagtgcca aaaatctttg agggtcttgg acgcgacgag 24000aagcgcgcgg caaacgctct gcaacaggaa aacagcgaaa atgaaagtca ctctggagtg 24060ttggtggaac tcgagggtga caacgcgcgc ctagccgtac taaaacgcag catcgaggtc 24120acccactttg cctacccggc acttaaccta ccccccaagg tcatgagcac agtcatgagt 24180gagctgatcg tgcgccgtgc gcagcccctg gagagggatg caaatttgca agaacaaaca 24240gaggagggcc tacccgcagt tggcgacgag cagctagcgc gctggcttca aacgcgcgag 24300cctgccgact tggaggagcg acgcaaacta atgatggccg cagtgctcgt taccgtggag 24360cttgagtgca tgcagcggtt ctttgctgac ccggagatgc agcgcaagct agaggaaaca 24420ttgcactaca cctttcgaca gggctacgta cgccaggcct gcaagatctc caacgtggag 24480ctctgcaacc tggtctccta ccttggaatt ttgcacgaaa accgccttgg gcaaaacgtg 24540cttcattcca cgctcaaggg cgaggcgcgc cgcgactacg tccgcgactg cgtttactta 24600tttctatgct acacctggca gacggccatg ggcgtttggc agcagtgctt ggaggagtgc 24660aacctcaagg agctgcagaa actgctaaag caaaacttga aggacctatg gacggccttc 24720aacgagcgct ccgtggccgc gcacctggcg gacatcattt tccccgaacg cctgcttaaa 24780accctgcaac agggtctgcc agacttcacc agtcaaagca tgttgcagaa ctttaggaac 24840tttatcctag agcgctcagg aatcttgccc gccacctgct gtgcacttcc tagcgacttt 24900gtgcccatta agtaccgcga atgccctccg ccgctttggg gccactgcta ccttctgcag 24960ctagccaact accttgccta ccactctgac ataatggaag acgtgagcgg tgacggtcta 25020ctggagtgtc actgtcgctg caacctatgc accccgcacc gctccctggt ttgcaattcg 25080cagctgctta acgaaagtca aattatcggt acctttgagc tgcagggtcc ctcgcctgac 25140gaaaagtccg cggctccggg gttgaaactc actccggggc tgtggacgtc ggcttacctt 25200cgcaaatttg tacctgagga ctaccacgcc cacgagatta ggttctacga agaccaatcc 25260cgcccgccta atgcggagct taccgcctgc gtcattaccc agggccacat tcttggccaa 25320ttgcaagcca tcaacaaagc ccgccaagag tttctgctac gaaagggacg gggggtttac 25380ttggaccccc agtccggcga ggagctcaac ccaatccccc cgccgccgca gccctatcag 25440cagcagccgc gggcccttgc ttcccaggat ggcacccaaa aagaagctgc agctgccgcc 25500gccacccacg gacgaggagg aatactggga cagtcaggca gaggaggttt tggacgagga 25560ggaggaggac atgatggaag actgggagag cctagacgag gaagcttccg aggtcgaaga 25620ggtgtcagac gaaacaccgt caccctcggt cgcattcccc tcgccggcgc cccagaaatc 25680ggcaaccggt tccagcatgg ctacaacctc cgctcctcag gcgccgccgg cactgcccgt 25740tcgccgaccc aaccgtagat gggacaccac tggaaccagg gccggtaagt ccaagcagcc 25800gccgccgtta gcccaagagc aacaacagcg ccaaggctac cgctcatggc gcgggcacaa 25860gaacgccata gttgcttgct tgcaagactg tgggggcaac atctccttcg cccgccgctt 25920tcttctctac catcacggcg tggccttccc ccgtaacatc ctgcattact accgtcatct 25980ctacagccca tactgcaccg gcggcagcgg cagcaacagc agcggccaca cagaagcaaa 26040ggcgaccgga tagcaagact ctgacaaagc ccaagaaatc cacagcggcg gcagcagcag 26100gaggaggagc gctgcgtctg gcgcccaacg aacccgtatc gacccgcgag cttagaaaca 26160ggatttttcc cactctgtat gctatatttc aacagagcag gggccaagaa caagagctga 26220aaataaaaaa caggtctctg cgatccctca cccgcagctg cctgtatcac aaaagcgaag 26280atcagcttcg gcgcacgctg gaagacgcgg aggctctctt cagtaaatac tgcgcgctga 26340ctcttaagga ctagtttcgc gccctttctc aaatttaagc gcgaaaacta cgtcatctcc 26400agcggccaca cccggcgcca gcacctgttg tcagcgccat tatgagcaag gaaattccca 26460cgccctacat gtggagttac cagccacaaa tgggacttgc ggctggagct gcccaagact 26520actcaacccg aataaactac atgagcgcgg gaccccacat gatatcccgg gtcaacggaa 26580tacgcgccca ccgaaaccga attctcctgg aacaggcggc tattaccacc acacctcgta 26640ataaccttaa tccccgtagt tggcccgctg ccctggtgta ccaggaaagt cccgctccca 26700ccactgtggt acttcccaga gacgcccagg ccgaagttca gatgactaac tcaggggcgc 26760agcttgcggg cggctttcgt cacagggtgc ggtcgcccgg gcagggtata actcacctga 26820caatcagagg gcgaggtatt cagctcaacg acgagtcggt gagctcctcg cttggtctcc 26880gtccggacgg gacatttcag atcggcggcg ccggccgctc ttcattcacg cctcgtcagg 26940caatcctaac tctgcagacc tcgtcctctg agccgcgctc tggaggcatt ggaactctgc 27000aatttattga ggagtttgtg ccatcggtct actttaaccc cttctcggga cctcccggcc 27060actatccgga tcaatttatt cctaactttg acgcggtaaa ggactcggcg gacggctacg 27120actgaatgtt aagtggagag gcagagcaac tgcgcctgaa acacctggtc cactgtcgcc 27180gccacaagtg ctttgcccgc gactccggtg agttttgcta ctttgaattg cccgaggatc 27240atatcgaggg cccggcgcac ggcgtccggc ttaccgccca gggagagctt gcccgtagcc 27300tgattcggga gtttacccag cgccccctgc tagttgagcg ggacagggga ccctgtgttc 27360tcactgtgat ttgcaactgt cctaaccctg gattacatca agatctttgt tgccatctct 27420gtgctgagta taataaatac agaaattaaa atatactggg gctcctatcg ccatcctgta 27480aacgccaccg tcttcacccg cccaagcaaa ccaaggcgaa ccttacctgg tacttttaac 27540atctctccct ctgtgattta caacagtttc aacccagacg gagtgagtct acgagagaac 27600ctctccgagc tcagctactc catcagaaaa aacaccaccc tccttacctg ccgggaacgt 27660acgagtgcgt caccggccgc tgcaccacac ctaccgcctg accgtaaacc agactttttc 27720cggacagacc tcaataactc tgtttaccag aacaggaggt gagcttagaa aacccttagg 27780gtattaggcc aaaggcgcag ctactgtggg gtttatgaac aattcaagca actctacggg 27840ctattctaat tcaggtttct ctagaatcgg ggttggggtt attctctgtc ttgtgattct 27900ctttattctt atactaacgc ttctctgcct aaggctcgcc gcctgctgtg tgcacatttg 27960catttattgt cagcttttta aacgctgggg tcgccaccca agatgattag gtacataatc 28020ctaggtttac tcacccttgc gtcagcccac ggtaccaccc aaaaggtgga ttttaaggag 28080ccagcctgta atgttacatt cgcagctgaa gctaatgagt gcaccactct tataaaatgc 28140accacagaac atgaaaagct gcttattcgc cacaaaaaca aaattggcaa gtatgctgtt 28200tatgctattt ggcagccagg tgacactaca gagtataatg ttacagtttt ccagggtaaa 28260agtcataaaa cttttatgta tacttttcca ttttatgaaa tgtgcgacat taccatgtac 28320atgagcaaac agtataagtt gtggccccca caaaattgtg tggaaaacac tggcactttc 28380tgctgcactg ctatgctaat tacagtgctc gctttggtct gtaccctact ctatattaaa 28440tacaaaagca gacgcagctt tattgaggaa aagaaaatgc cttaatttac taagttacaa 28500agctaatgtc accactaact gctttactcg ctgcttgcaa aacaaattca aaaagttagc 28560attataatta gaataggatt taaacccccc ggtcatttcc tgctcaatac cattcccctg 28620aacaattgac tctatgtggg atatgctcca gcgctacaac cttgaagtca ggcttcctgg 28680atgtcagcat ctgactttgg ccagcacctg tcccgcggat ttgttccagt ccaactacag 28740cgacccaccc taacagagat gaccaacaca accaacgcgg ccgccgctac cggacttaca 28800tctaccacaa atacacccca agtttctgcc tttgtcaata actgggataa cttgggcatg 28860tggtggttct ccatagcgct tatgtttgta tgccttatta ttatgtggct catctgctgc 28920ctaaagcgca aacgcgcccg accacccatc tatagtccca tcattgtgct acacccaaac 28980aatgatggaa tccatagatt ggacggactg aaacacatgt tcttttctct tacagtatga 29040ttaaatgaga catgattcct cgagttttta tattactgac ccttgttgcg ctttttttgt 29100gcgtgctcca cattggctgc ggtttctcac atcgaagtag actgcattcc agccttcaca 29160gtctatttgc tttacggatt tgtcaccctc acgctcatct gcagcctcat cactgtggtc 29220atcgccttta tccagtgcat tgactgggtc tgtgtgcgct ttgcatatct cagacaccat 29280ccccagtaca gggacaggac tatagctgag cttcttagaa ttctttaatt atgaaattta 29340ctgtgacttt tctgctgatt atttgcaccc tatctgcgtt ttgttccccg acctccaagc 29400ctcaaagaca tatatcatgc agattcactc gtatatggaa tattccaagt tgctacaatg 29460aaaaaagcga tctttccgaa gcctggttat atgcaatcat ctctgttatg gtgttctgca 29520gtaccatctt agccctagct atatatccct accttgacat tggctggaac gcaatagatg 29580ccatgaacca cccaactttc cccgcgcccg ctatgcttcc actgcaacaa gttgttgccg 29640gcggctttgt cccagccaat cagcctcgcc caccttctcc cacccccact gaaatcagct 29700actttaatct aacaggagga gatgactgac accctagatc tagaaatgga cggaattatt 29760acagagcagc gcctgctaga aagacgcagg gcagcggccg agcaacagcg catgaatcaa 29820gagctccaag acatggttaa cttgcaccag tgcaaaaggg gtatcttttg tctggtaaag 29880caggccaaag tcacctacga cagtaatacc accggacacc gccttagcta caagttgcca 29940accaagcgtc agaaattggt ggtcatggtg ggagaaaagc ccattaccat aactcagcac 30000tcggtagaaa ccgaaggctg cattcactca ccttgtcaag gacctgagga tctctgcacc 30060cttattaaga ccctgtgcgg tctcaaagat cttattccct ttaactaata aaaaaaaata 30120ataaagcatc acttacttaa aatcagttag caaatttctg tccagtttat tcagcagcac 30180ctccttgccc tcctcccagc tctggtattg cagcttcctc ctggctgcaa actttctcca 30240caatctaaat ggaatgtcag tttcctcctg ttcctgtcca tccgcaccca ctatcttcat 30300gttgttgcag atgaagcgcg caagaccgtc tgaagatacc ttcaaccccg tgtatccata 30360tgacacggaa accggtcctc caactgtgcc ttttcttact cctccctttg tatcccccaa 30420tgggtttcaa gagagtcccc ctggggtact ctctttgcgc ctatccgaac ctctagttac 30480ctccaatggc atgcttgcgc tcaaaatggg caacggcctc tctctggacg aggccggcaa 30540ccttacctcc caaaatgtaa ccactgtgag cccacctctc aaaaaaacca agtcaaacat 30600aaacctggaa atatctgcac ccctcacagt tacctcagaa gccctaactg tggctgccgc 30660cgcacctcta atggtcgcgg gcaacacact caccatgcaa tcacaggccc cgctaaccgt 30720gcacgactcc aaacttagca ttgccaccca aggacccctc acagtgtcag aaggaaagct 30780agccctgcaa acatcaggcc ccctcaccac caccgatagc agtaccctta ctatcactgc 30840ctcaccccct ctaactactg ccactggtag cttgggcatt gacttgaaag agcccattta 30900tacacaaaat ggaaaactag gactaaagta cggggctcct ttgcatgtaa cagacgacct 30960aaacactttg accgtagcaa ctggtccagg tgtgactatt aataatactt ccttgcaaac 31020taaagttact ggagccttgg gttttgattc acaaggcaat atgcaactta atgtagcagg 31080aggactaagg attgattctc aaaacagacg ccttatactt gatgttagtt atccgtttga 31140tgctcaaaac caactaaatc taagactagg acagggccct ctttttataa actcagccca 31200caacttggat attaactaca acaaaggcct ttacttgttt acagcttcaa acaattccaa 31260aaagcttgag

gttaacctaa gcactgccaa ggggttgatg tttgacgcta cagccatagc 31320cattaatgca ggagatgggc ttgaatttgg ttcacctaat gcaccaaaca caaatcccct 31380caaaacaaaa attggccatg gcctagaatt tgattcaaac aaggctatgg ttcctaaact 31440aggaactggc cttagttttg acagcacagg tgccattaca gtaggaaaca aaaataatga 31500taagctaact ttgtggacca caccagctcc atctcctaac tgtagactaa atgcagagaa 31560agatgctaaa ctcactttgg tcttaacaaa atgtggcagt caaatacttg ctacagtttc 31620agttttggct gttaaaggca gtttggctcc aatatctgga acagttcaaa gtgctcatct 31680tattataaga tttgacgaaa atggagtgct actaaacaat tccttcctgg acccagaata 31740ttggaacttt agaaatggag atcttactga aggcacagcc tatacaaacg ctgttggatt 31800tatgcctaac ctatcagctt atccaaaatc tcacggtaaa actgccaaaa gtaacattgt 31860cagtcaagtt tacttaaacg gagacaaaac taaacctgta acactaacca ttacactaaa 31920cggtacacag gaaacaggag acacaactcc aagtgcatac tctatgtcat tttcatggga 31980ctggtctggc cacaactaca ttaatgaaat atttgccaca tcctcttaca ctttttcata 32040cattgcccaa gaataaagaa tcgtttgtgt tatgtttcaa cgtgtttatt tttcaattgc 32100agaaaatttc aagtcatttt tcattcagta gtatagcccc accaccacat agcttataca 32160gatcaccgta ccttaatcaa actcacagaa ccctagtatt caacctgcca cctccctccc 32220aacacacaga gtacacagtc ctttctcccc ggctggcctt aaaaagcatc atatcatggg 32280taacagacat attcttaggt gttatattcc acacggtttc ctgtcgagcc aaacgctcat 32340cagtgatatt aataaactcc ccgggcagct cacttaagtt catgtcgctg tccagctgct 32400gagccacagg ctgctgtcca acttgcggtt gcttaacggg cggcgaagga gaagtccacg 32460cctacatggg ggtagagtca taatcgtgca tcaggatagg gcggtggtgc tgcagcagcg 32520cgcgaataaa ctgctgccgc cgccgctccg tcctgcagga atacaacatg gcagtggtct 32580cctcagcgat gattcgcacc gcccgcagca taaggcgcct tgtcctccgg gcacagcagc 32640gcaccctgat ctcacttaaa tcagcacagt aactgcagca cagcaccaca atattgttca 32700aaatcccaca gtgcaaggcg ctgtatccaa agctcatggc ggggaccaca gaacccacgt 32760ggccatcata ccacaagcgc aggtagatta agtggcgacc cctcataaac acgctggaca 32820taaacattac ctcttttggc atgttgtaat tcaccacctc ccggtaccat ataaacctct 32880gattaaacat ggcgccatcc accaccatcc taaaccagct ggccaaaacc tgcccgccgg 32940ctatacactg cagggaaccg ggactggaac aatgacagtg gagagcccag gactcgtaac 33000catggatcat catgctcgtc atgatatcaa tgttggcaca acacaggcac acgtgcatac 33060acttcctcag gattacaagc tcctcccgcg ttagaaccat atcccaggga acaacccatt 33120cctgaatcag cgtaaatccc acactgcagg gaagacctcg cacgtaactc acgttgtgca 33180ttgtcaaagt gttacattcg ggcagcagcg gatgatcctc cagtatggta gcgcgggttt 33240ctgtctcaaa aggaggtaga cgatccctac tgtacggagt gcgccgagac aaccgagatc 33300gtgttggtcg tagtgtcatg ccaaatggaa cgccggacgt agtcatattt cctgaagcaa 33360aaccaggtgc gggcgtgaca aacagatctg cgtctccggt ctcgccgctt agatcgctct 33420gtgtagtagt tgtagtatat ccactctctc aaagcatcca ggcgccccct ggcttcgggt 33480tctatgtaaa ctccttcatg cgccgctgcc ctgataacat ccaccaccgc agaataagcc 33540acacccagcc aacctacaca ttcgttctgc gagtcacaca cgggaggagc gggaagagct 33600ggaagaacca tgtttttttt tttattccaa aagattatcc aaaacctcaa aatgaagatc 33660tattaagtga acgcgctccc ctccggtggc gtggtcaaac tctacagcca aagaacagat 33720aatggcattt gtaagatgtt gcacaatggc ttccaaaagg caaacggccc tcacgtccaa 33780gtggacgtaa aggctaaacc cttcagggtg aatctcctct ataaacattc cagcaccttc 33840aaccatgccc aaataattct catctcgcca ccttctcaat atatctctaa gcaaatcccg 33900aatattaagt ccggccattg taaaaatctg ctccagagcg ccctccacct tcagcctcaa 33960gcagcgaatc atgattgcaa aaattcaggt tcctcacaga cctgtataag attcaaaagc 34020ggaacattaa caaaaatacc gcgatcccgt aggtcccttc gcagggccag ctgaacataa 34080tcgtgcaggt ctgcacggac cagcgcggcc acttccccgc caggaaccat gacaaaagaa 34140cccacactga ttatgacacg catactcgga gctatgctaa ccagcgtagc cccgatgtaa 34200gcttgttgca tgggcggcga tataaaatgc aaggtgctgc tcaaaaaatc aggcaaagcc 34260tcgcgcaaaa aagaaagcac atcgtagtca tgctcatgca gataaaggca ggtaagctcc 34320ggaaccacca cagaaaaaga caccattttt ctctcaaaca tgtctgcggg tttctgcata 34380aacacaaaat aaaataacaa aaaaacattt aaacattaga agcctgtctt acaacaggaa 34440aaacaaccct tataagcata agacggacta cggccatgcc ggcgtgaccg taaaaaaact 34500ggtcaccgtg attaaaaagc accaccgaca gctcctcggt catgtccgga gtcataatgt 34560aagactcggt aaacacatca ggttgattca catcggtcag tgctaaaaag cgaccgaaat 34620agcccggggg aatacatacc cgcaggcgta gagacaacat tacagccccc ataggaggta 34680taacaaaatt aataggagag aaaaacacat aaacacctga aaaaccctcc tgcctaggca 34740aaatagcacc ctcccgctcc agaacaacat acagcgcttc cacagcggca gccataacag 34800tcagccttac cagtaaaaaa gaaaacctat taaaaaaaca ccactcgaca cggcaccagc 34860tcaatcagtc acagtgtaaa aaagggccaa gtgcagagcg agtatatata ggactaaaaa 34920atgacgtaac ggttaaagtc cacaaaaaac acccagaaaa ccgcacgcga acctacgccc 34980agaaacgaaa gccaaaaaac ccacaacttc ctcaaatcgt cacttccgtt ttcccacgtt 35040acgtcacttc ccattttaag aaaactacaa ttcccaacac atacaagtta ctccgcccta 35100aaacctacgt cacccgcccc gttcccacgc cccgcgccac gtcacaaact ccaccccctc 35160attatcatat tggcttcaat ccaaaataag gtatattatt gat 352031133093DNAArtificial sequenceEmpty Ad5 vector sequence (repeats included) 11catcatcaat aatatacctt attttggatt gaagccaata tgataatgag ggggtggagt 60ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg tagtagtgtg gcggaagtgt 120gatgttgcaa gtgtggcgga acacatgtaa gcgacggatg tggcaaaagt gacgtttttg 180gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg gttttaggcg gatgttgtag 240taaatttggg cgtaaccgag taagatttgg ccattttcgc gggaaaactg aataagagga 300agtgaaatct gaataatttt gtgttactca tagcgcgtaa tatttgtcta gggccgcggg 360gactttgacc gtttacgtgg agactcgccc aggtgttttt ctcaggtgtt ttccgcgttc 420cgggtcaaag ttggcgtttt attattatag tcagtacgtc tcgagcatgc atctaggcgg 480ccgcatggca gaaattcgcg aattcgctag cgttaacgga tcctctagac gagatccgaa 540cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 600taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 660tcatgtctag atctgtactg aaatgtgtgg gcgtggctta agggtgggaa agaatatata 720aggtgggggt cttatgtagt tttgtatctg ttttgcagca gccgccgccg ccatgagcac 780caactcgttt gatggaagca ttgtgagctc atatttgaca acgcgcatgc ccccatgggc 840cggggtgcgt cagaatgtga tgggctccag cattgatggt cgccccgtcc tgcccgcaaa 900ctctactacc ttgacctacg agaccgtgtc tggaacgccg ttggagactg cagcctccgc 960cgccgcttca gccgctgcag ccaccgcccg cgggattgtg actgactttg ctttcctgag 1020cccgcttgca agcagtgcag cttcccgttc atccgcccgc gatgacaagt tgacggctct 1080tttggcacaa ttggattctt tgacccggga acttaatgtc gtttctcagc agctgttgga 1140tctgcgccag caggtttctg ccctgaaggc ttcctcccct cccaatgcgg tttaaaacat 1200aaataaaaaa ccagactctg tttggatttg gatcaagcaa gtgtcttgct gtctttattt 1260aggggttttg cgcgcgcggt aggcccggga ccagcggtct cggtcgttga gggtcctgtg 1320tattttttcc aggacgtggt aaaggtgact ctggatgttc agatacatgg gcataagccc 1380gtctctgggg tggaggtagc accactgcag agcttcatgc tgcggggtgg tgttgtagat 1440gatccagtcg tagcaggagc gctgggcgtg gtgcctaaaa atgtctttca gtagcaagct 1500gattgccagg ggcaggccct tggtgtaagt gtttacaaag cggttaagct gggatgggtg 1560catacgtggg gatatgagat gcatcttgga ctgtattttt aggttggcta tgttcccagc 1620catatccctc cggggattca tgttgtgcag aaccaccagc acagtgtatc cggtgcactt 1680gggaaatttg tcatgtagct tagaaggaaa tgcgtggaag aacttggaga cgcccttgtg 1740acctccaaga ttttccatgc attcgtccat aatgatggca atgggcccac gggcggcggc 1800ctgggcgaag atatttctgg gatcactaac gtcatagttg tgttccagga tgagatcgtc 1860ataggccatt tttacaaagc gcgggcggag ggtgccagac tgcggtataa tggttccatc 1920cggcccaggg gcgtagttac cctcacagat ttgcatttcc cacgctttga gttcagatgg 1980ggggatcatg tctacctgcg gggcgatgaa gaaaacggtt tccggggtag gggagatcag 2040ctgggaagaa agcaggttcc tgagcagctg cgacttaccg cagccggtgg gcccgtaaat 2100cacacctatt accggctgca actggtagtt aagagagctg cagctgccgt catccctgag 2160caggggggcc acttcgttaa gcatgtccct gactcgcatg ttttccctga ccaaatccgc 2220cagaaggcgc tcgccgccca gcgatagcag ttcttgcaag gaagcaaagt ttttcaacgg 2280tttgagaccg tccgccgtag gcatgctttt gagcgtttga ccaagcagtt ccaggcggtc 2340ccacagctcg gtcacctgct ctacggcatc tcgatccagc atatctcctc gtttcgcggg 2400ttggggcggc tttcgctgta cggcagtagt cggtgctcgt ccagacgggc cagggtcatg 2460tctttccacg ggcgcagggt cctcgtcagc gtagtctggg tcacggtgaa ggggtgcgct 2520ccgggctgcg cgctggccag ggtgcgcttg aggctggtcc tgctggtgct gaagcgctgc 2580cggtcttcgc cctgcgcgtc ggccaggtag catttgacca tggtgtcata gtccagcccc 2640tccgcggcgt ggcccttggc gcgcagcttg cccttggagg aggcgccgca cgaggggcag 2700tgcagacttt tgagggcgta gagcttgggc gcgagaaata ccgattccgg ggagtaggca 2760tccgcgccgc aggccccgca gacggtctcg cattccacga gccaggtgag ctctggccgt 2820tcggggtcaa aaaccaggtt tcccccatgc tttttgatgc gtttcttacc tctggtttcc 2880atgagccggt gtccacgctc ggtgacgaaa aggctgtccg tgtccccgta tacagacttg 2940agaggcctgt cctcgagcgg tgttccgcgg tcctcctcgt atagaaactc ggaccactct 3000gagacaaagg ctcgcgtcca ggccagcacg aaggaggcta agtgggaggg gtagcggtcg 3060ttgtccacta gggggtccac tcgctccagg gtgtgaagac acatgtcgcc ctcttcggca 3120tcaaggaagg tgattggttt gtaggtgtag gccacgtgac cgggtgttcc tgaagggggg 3180ctataaaagg gggtgggggc gcgttcgtcc tcactctctt ccgcatcgct gtctgcgagg 3240gccagctgtt ggggtgagta ctccctctga aaagcgggca tgacttctgc gctaagattg 3300tcagtttcca aaaacgagga ggatttgata ttcacctggc ccgcggtgat gcctttgagg 3360gtggccgcat ccatctggtc agaaaagaca atctttttgt tgtcaagctt ggtggcaaac 3420gacccgtaga gggcgttgga cagcaacttg gcgatggagc gcagggtttg gtttttgtcg 3480cgatcggcgc gctccttggc cgcgatgttt agctgcacgt attcgcgcgc aacgcaccgc 3540cattcgggaa agacggtggt gcgctcgtcg ggcaccaggt gcacgcgcca accgcggttg 3600tgcagggtga caaggtcaac gctggtggct acctctccgc gtaggcgctc gttggtccag 3660cagaggcggc cgcccttgcg cgagcagaat ggcggtaggg ggtctagctg cgtctcgtcc 3720ggggggtctg cgtccacggt aaagaccccg ggcagcaggc gcgcgtcgaa gtagtctatc 3780ttgcatcctt gcaagtctag cgcctgctgc catgcgcggg cggcaagcgc gcgctcgtat 3840gggttgagtg ggggacccca tggcatgggg tgggtgagcg cggaggcgta catgccgcaa 3900atgtcgtaaa cgtagagggg ctctctgagt attccaagat atgtagggta gcatcttcca 3960ccgcggatgc tggcgcgcac gtaatcgtat agttcgtgcg agggagcgag gaggtcggga 4020ccgaggttgc tacgggcggg ctgctctgct cggaagacta tctgcctgaa gatggcatgt 4080gagttggatg atatggttgg acgctggaag acgttgaagc tggcgtctgt gagacctacc 4140gcgtcacgca cgaaggaggc gtaggagtcg cgcagcttgt tgaccagctc ggcggtgacc 4200tgcacgtcta gggcgcagta gtccagggtt tccttgatga tgtcatactt atcctgtccc 4260ttttttttcc acagctcgcg gttgaggaca aactcttcgc ggtctttcca gtactcttgg 4320atcggaaacc cgtcggcctc cgaacggtaa gagcctagca tgtagaactg gttgacggcc 4380tggtaggcgc agcatccctt ttctacgggt agcgcgtatg cctgcgcggc cttccggagc 4440gaggtgtggg tgagcgcaaa ggtgtccctg accatgactt tgaggtactg gtatttgaag 4500tcagtgtcgt cgcatccgcc ctgctcccag agcaaaaagt ccgtgcgctt tttggaacgc 4560ggatttggca gggcgaaggt gacatcgttg aagagtatct ttcccgcgcg aggcataaag 4620ttgcgtgtga tgcggaaggg tcccggcacc tcggaacggt tgttaattac ctgggcggcg 4680agcacgatct cgtcaaagcc gttgatgttg tggcccacaa tgtaaagttc caagaagcgc 4740gggatgccct tgatggaagg caatttttta agttcctcgt aggtgagctc ttcaggggag 4800ctgagcccgt gctctgaaag ggcccagtct gcaagatgag ggttggaagc gacgaatgag 4860ctccacaggt cacgggccat tagcatttgc aggtggtcgc gaaaggtcct aaactggcga 4920cctatggcca ttttttctgg ggtgatgcag tagaaggtaa gcgggtcttg ttcccagcgg 4980tcccatccaa ggttcgcggc taggtctcgc gcggcagtca ctagaggctc atctccgccg 5040aacttcatga ccagcatgaa gggcacgagc tgcttcccaa aggcccccat ccaagtatag 5100gtctctacat cgtaggtgac aaagagacgc tcggtgcgag gatgcgagcc gatcgggaag 5160aactggatct cccgccacca attggaggag tggctattga tgtggtgaaa gtagaagtcc 5220ctgcgacggg ccgaacactc gtgctggctt ttgtaaaaac gtgcgcagta ctggcagcgg 5280tgcacgggct gtacatcctg cacgaggttg acctgacgac cgcgcacaag gaagcagagt 5340gggaatttga gcccctcgcc tggcgggttt ggctggtggt cttctacttc ggctgcttgt 5400ccttgaccgt ctggctgctc gaggggagtt acggtggatc ggaccaccac gccgcgcgag 5460cccaaagtcc agatgtccgc gcgcggcggt cggagcttga tgacaacatc gcgcagatgg 5520gagctgtcca tggtctggag ctcccgcggc gtcaggtcag gcgggagctc ctgcaggttt 5580acctcgcata gacgggtcag ggcgcgggct agatccaggt gatacctaat ttccaggggc 5640tggttggtgg cggcgtcgat ggcttgcaag aggccgcatc cccgcggcgc gactacggta 5700ccgcgcggcg ggcggtgggc cgcgggggtg tccttggatg atgcatctaa aagcggtgac 5760gcgggcgagc ccccggaggt agggggggct ccggacccgc cgggagaggg ggcaggggca 5820cgtcggcgcc gcgcgcgggc aggagctggt gctgcgcgcg taggttgctg gcgaacgcga 5880cgacgcggcg gttgatctcc tgaatctggc gcctctgcgt gaagacgacg ggcccggtga 5940gcttgaacct gaaagagagt tcgacagaat caatttcggt gtcgttgacg gcggcctggc 6000gcaaaatctc ctgcacgtct cctgagttgt cttgataggc gatctcggcc atgaactgct 6060cgatctcttc ctcctggaga tctccgcgtc cggctcgctc cacggtggcg gcgaggtcgt 6120tggaaatgcg ggccatgagc tgcgagaagg cgttgaggcc tccctcgttc cagacgcggc 6180tgtagaccac gcccccttcg gcatcgcggg cgcgcatgac cacctgcgcg agattgagct 6240ccacgtgccg ggcgaagacg gcgtagtttc gcaggcgctg aaagaggtag ttgagggtgg 6300tggcggtgtg ttctgccacg aagaagtaca taacccagcg tcgcaacgtg gattcgttga 6360tatcccccaa ggcctcaagg cgctccatgg cctcgtagaa gtccacggcg aagttgaaaa 6420actgggagtt gcgcgccgac acggttaact cctcctccag aagacggatg agctcggcga 6480cagtgtcgcg cacctcgcgc tcaaaggcta caggggcctc ttcttcttct tcaatctcct 6540cttccataag ggcctcccct tcttcttctt ctggcggcgg tgggggaggg gggacacggc 6600ggcgacgacg gcgcaccggg aggcggtcga caaagcgctc gatcatctcc ccgcggcgac 6660ggcgcatggt ctcggtgacg gcgcggccgt tctcgcgggg gcgcagttgg aagacgccgc 6720ccgtcatgtc ccggttatgg gttggcgggg ggctgccatg cggcagggat acggcgctaa 6780cgatgcatct caacaattgt tgtgtaggta ctccgccgcc gagggacctg agcgagtccg 6840catcgaccgg atcggaaaac ctctcgagaa aggcgtctaa ccagtcacag tcgcaaggta 6900ggctgagcac cgtggcgggc ggcagcgggc ggcggtcggg gttgtttctg gcggaggtgc 6960tgctgatgat gtaattaaag taggcggtct tgagacggcg gatggtcgac agaagcacca 7020tgtccttggg tccggcctgc tgaatgcgca ggcggtcggc catgccccag gcttcgtttt 7080gacatcggcg caggtctttg tagtagtctt gcatgagcct ttctaccggc acttcttctt 7140ctccttcctc ttgtcctgca tctcttgcat ctatcgctgc ggcggcggcg gagtttggcc 7200gtaggtggcg ccctcttcct cccatgcgtg tgaccccgaa gcccctcatc ggctgaagca 7260gggctaggtc ggcgacaacg cgctcggcta atatggcctg ctgcacctgc gtgagggtag 7320actggaagtc atccatgtcc acaaagcggt ggtatgcgcc cgtgttgatg gtgtaagtgc 7380agttggccat aacggaccag ttaacggtct ggtgacccgg ctgcgagagc tcggtgtacc 7440tgagacgcga gtaagccctc gagtcaaata cgtagtcgtt gcaagtccgc accaggtact 7500ggtatcccac caaaaagtgc ggcggcggct ggcggtagag gggccagcgt agggtggccg 7560gggctccggg ggcgagatct tccaacataa ggcgatgata tccgtagatg tacctggaca 7620tccaggtgat gccggcggcg gtggtggagg cgcgcggaaa gtcgcggacg cggttccaga 7680tgttgcgcag cggcaaaaag tgctccatgg tcgggacgct ctggccggtc aggcgcgcgc 7740aatcgttgac gctctagacc gtgcaaaagg agagcctgta agcgggcact cttccgtggt 7800ctggtggata aattcgcaag ggtatcatgg cggacgaccg gggttcgagc cccgtatccg 7860gccgtccgcc gtgatccatg cggttaccgc ccgcgtgtcg aacccaggtg tgcgacgtca 7920gacaacgggg gagtgctcct tttggcttcc ttccaggcgc ggcggctgct gcgctagctt 7980ttttggccac tggccgcgcg cagcgtaagc ggttaggctg gaaagcgaaa gcattaagtg 8040gctcgctccc tgtagccgga gggttatttt ccaagggttg agtcgcggga cccccggttc 8100gagtctcgga ccggccggac tgcggcgaac gggggtttgc ctccccgtca tgcaagaccc 8160cgcttgcaaa ttcctccgga aacagggacg agcccctttt ttgcttttcc cagatgcatc 8220cggtgctgcg gcagatgcgc ccccctcctc agcagcggca agagcaagag cagcggcaga 8280catgcagggc accctcccct cctcctaccg cgtcaggagg ggcgacatcc gcggttgacg 8340cggcagcaga tggtgattac gaacccccgc ggcgccgggc ccggcactac ctggacttgg 8400aggagggcga gggcctggcg cggctaggag cgccctctcc tgagcggcac ccaagggtgc 8460agctgaagcg tgatacgcgt gaggcgtacg tgccgcggca gaacctgttt cgcgaccgcg 8520agggagagga gcccgaggag atgcgggatc gaaagttcca cgcagggcgc gagctgcggc 8580atggcctgaa tcgcgagcgg ttgctgcgcg aggaggactt tgagcccgac gcgcgaaccg 8640ggattagtcc cgcgcgcgca cacgtggcgg ccgccgacct ggtaaccgca tacgagcaga 8700cggtgaacca ggagattaac tttcaaaaaa gctttaacaa ccacgtgcgt acgcttgtgg 8760cgcgcgagga ggtggctata ggactgatgc atctgtggga ctttgtaagc gcgctggagc 8820aaaacccaaa tagcaagccg ctcatggcgc agctgttcct tatagtgcag cacagcaggg 8880acaacgaggc attcagggat gcgctgctaa acatagtaga gcccgagggc cgctggctgc 8940tcgatttgat aaacatcctg cagagcatag tggtgcagga gcgcagcttg agcctggctg 9000acaaggtggc cgccatcaac tattccatgc ttagcctggg caagttttac gcccgcaaga 9060tataccatac cccttacgtt cccatagaca aggaggtaaa gatcgagggg ttctacatgc 9120gcatggcgct gaaggtgctt accttgagcg acgacctggg cgtttatcgc aacgagcgca 9180tccacaaggc cgtgagcgtg agccggcggc gcgagctcag cgaccgcgag ctgatgcaca 9240gcctgcaaag ggccctggct ggcacgggca gcggcgatag agaggccgag tcctactttg 9300acgcgggcgc tgacctgcgc tgggccccaa gccgacgcgc cctggaggca gctggggccg 9360gacctgggct ggcggtggca cccgcgcgcg ctggcaacgt cggcggcgtg gaggaatatg 9420acgaggacga tgagtacgag ccagaggacg gcgagtacta agcggtgatg tttctgatca 9480gatgatgcaa gacgcaacgg acccggcggt gcgggcggcg ctgcagagcc agccgtccgg 9540ccttaactcc acggacgact ggcgccaggt catggaccgc atcatgtcgc tgactgcgcg 9600caatcctgac gcgttccggc agcagccgca ggccaaccgg ctctccgcaa ttctggaagc 9660ggtggtcccg gcgcgcgcaa accccacgca cgagaaggtg ctggcgatcg taaacgcgct 9720ggccgaaaac agggccatcc ggcccgacga ggccggcctg gtctacgacg cgctgcttca 9780gcgcgtggct cgttacaaca gcggcaacgt gcagaccaac ctggaccggc tggtggggga 9840tgtgcgcgag gccgtggcgc agcgtgagcg cgcgcagcag cagggcaacc tgggctccat 9900ggttgcacta aacgccttcc tgagtacaca gcccgccaac gtgccgcggg gacaggagga 9960ctacaccaac tttgtgagcg cactgcggct aatggtgact gagacaccgc aaagtgaggt 10020gtaccagtct gggccagact attttttcca gaccagtaga caaggcctgc agaccgtaaa 10080cctgagccag gctttcaaaa acttgcaggg gctgtggggg gtgcgggctc ccacaggcga 10140ccgcgcgacc gtgtctagct tgctgacgcc caactcgcgc ctgttgctgc tgctaatagc 10200gcccttcacg gacagtggca gcgtgtcccg ggacacatac ctaggtcact tgctgacact 10260gtaccgcgag gccataggtc aggcgcatgt ggacgagcat actttccagg agattacaag 10320tgtcagccgc gcgctggggc aggaggacac gggcagcctg gaggcaaccc taaactacct 10380gctgaccaac cggcggcaga agatcccctc gttgcacagt ttaaacagcg aggaggagcg 10440cattttgcgc tacgtgcagc agagcgtgag ccttaacctg atgcgcgacg gggtaacgcc 10500cagcgtggcg ctggacatga ccgcgcgcaa catggaaccg ggcatgtatg cctcaaaccg 10560gccgtttatc aaccgcctaa tggactactt gcatcgcgcg gccgccgtga accccgagta 10620tttcaccaat gccatcttga acccgcactg gctaccgccc cctggtttct acaccggggg 10680attcgaggtg cccgagggta acgatggatt cctctgggac gacatagacg acagcgtgtt 10740ttccccgcaa ccgcagaccc tgctagagtt gcaacagcgc gagcaggcag aggcggcgct 10800gcgaaaggaa agcttccgca ggccaagcag cttgtccgat ctaggcgctg cggccccgcg 10860gtcagatgct agtagcccat ttccaagctt gatagggtct cttaccagca ctcgcaccac 10920ccgcccgcgc ctgctgggcg aggaggagta cctaaacaac tcgctgctgc agccgcagcg 10980cgaaaaaaac ctgcctccgg catttcccaa caacgggata gagagcctag

tggacaagat 11040gagtagatgg aagacgtacg cgcaggagca cagggacgtg ccaggcccgc gcccgcccac 11100ccgtcgtcaa aggcacgacc gtcagcgggg tctggtgtgg gaggacgatg actcggcaga 11160cgacagcagc gtcctggatt tgggagggag tggcaacccg tttgcgcacc ttcgccccag 11220gctggggaga atgttttaaa aaaaaaaaaa gcatgatgca aaataaaaaa ctcaccaagg 11280ccatggcacc gagcgttggt tttcttgtat tccccttagt atgcggcgcg cggcgatgta 11340tgaggaaggt cctcctccct cctacgagag tgtggtgagc gcggcgccag tggcggcggc 11400gctgggttct cccttcgatg ctcccctgga cccgccgttt gtgcctccgc ggtacctgcg 11460gcctaccggg gggagaaaca gcatccgtta ctctgagttg gcacccctat tcgacaccac 11520ccgtgtgtac ctggtggaca acaagtcaac ggatgtggca tccctgaact accagaacga 11580ccacagcaac tttctgacca cggtcattca aaacaatgac tacagcccgg gggaggcaag 11640cacacagacc atcaatcttg acgaccggtc gcactggggc ggcgacctga aaaccatcct 11700gcataccaac atgccaaatg tgaacgagtt catgtttacc aataagttta aggcgcgggt 11760gatggtgtcg cgcttgccta ctaaggacaa tcaggtggag ctgaaatacg agtgggtgga 11820gttcacgctg cccgagggca actactccga gaccatgacc atagacctta tgaacaacgc 11880gatcgtggag cactacttga aagtgggcag acagaacggg gttctggaaa gcgacatcgg 11940ggtaaagttt gacacccgca acttcagact ggggtttgac cccgtcactg gtcttgtcat 12000gcctggggta tatacaaacg aagccttcca tccagacatc attttgctgc caggatgcgg 12060ggtggacttc acccacagcc gcctgagcaa cttgttgggc atccgcaagc ggcaaccctt 12120ccaggagggc tttaggatca cctacgatga tctggagggt ggtaacattc ccgcactgtt 12180ggatgtggac gcctaccagg cgagcttgaa agatgacacc gaacagggcg ggggtggcgc 12240aggcggcagc aacagcagtg gcagcggcgc ggaagagaac tccaacgcgg cagccgcggc 12300aatgcagccg gtggaggaca tgaacgatca tgccattcgc ggcgacacct ttgccacacg 12360ggctgaggag aagcgcgctg aggccgaagc agcggccgaa gctgccgccc ccgctgcgca 12420acccgaggtc gagaagcctc agaagaaacc ggtgatcaaa cccctgacag aggacagcaa 12480gaaacgcagt tacaacctaa taagcaatga cagcaccttc acccagtacc gcagctggta 12540ccttgcatac aactacggcg accctcagac cggaatccgc tcatggaccc tgctttgcac 12600tcctgacgta acctgcggct cggagcaggt ctactggtcg ttgccagaca tgatgcaaga 12660ccccgtgacc ttccgctcca cgcgccagat cagcaacttt ccggtggtgg gcgccgagct 12720gttgcccgtg cactccaaga gcttctacaa cgaccaggcc gtctactccc aactcatccg 12780ccagtttacc tctctgaccc acgtgttcaa tcgctttccc gagaaccaga ttttggcgcg 12840cccgccagcc cccaccatca ccaccgtcag tgaaaacgtt cctgctctca cagatcacgg 12900gacgctaccg ctgcgcaaca gcatcggagg agtccagcga gtgaccatta ctgacgccag 12960acgccgcacc tgcccctacg tttacaaggc cctgggcata gtctcgccgc gcgtcctatc 13020gagccgcact ttttgagcaa gcatgtccat ccttatatcg cccagcaata acacaggctg 13080gggcctgcgc ttcccaagca agatgtttgg cggggccaag aagcgctccg accaacaccc 13140agtgcgcgtg cgcgggcact accgcgcgcc ctggggcgcg cacaaacgcg gccgcactgg 13200gcgcaccacc gtcgatgacg ccatcgacgc ggtggtggag gaggcgcgca actacacgcc 13260cacgccgcca ccagtgtcca cagtggacgc ggccattcag accgtggtgc gcggagcccg 13320gcgctatgct aaaatgaaga gacggcggag gcgcgtagca cgtcgccacc gccgccgacc 13380cggcactgcc gcccaacgcg cggcggcggc cctgcttaac cgcgcacgtc gcaccggccg 13440acgggcggcc atgcgggccg ctcgaaggct ggccgcgggt attgtcactg tgccccccag 13500gtccaggcga cgagcggccg ccgcagcagc cgcggccatt agtgctatga ctcagggtcg 13560caggggcaac gtgtattggg tgcgcgactc ggttagcggc ctgcgcgtgc ccgtgcgcac 13620ccgccccccg cgcaactaga ttgcaagaaa aaactactta gactcgtact gttgtatgta 13680tccagcggcg gcggcgcgca acgaagctat gtccaagcgc aaaatcaaag aagagatgct 13740ccaggtcatc gcgccggaga tctatggccc cccgaagaag gaagagcagg attacaagcc 13800ccgaaagcta aagcgggtca aaaagaaaaa gaaagatgat gatgatgaac ttgacgacga 13860ggtggaactg ctgcacgcta ccgcgcccag gcgacgggta cagtggaaag gtcgacgcgt 13920aaaacgtgtt ttgcgacccg gcaccaccgt agtctttacg cccggtgagc gctccacccg 13980cacctacaag cgcgtgtatg atgaggtgta cggcgacgag gacctgcttg agcaggccaa 14040cgagcgcctc ggggagtttg cctacggaaa gcggcataag gacatgctgg cgttgccgct 14100ggacgagggc aacccaacac ctagcctaaa gcccgtaaca ctgcagcagg tgctgcccgc 14160gcttgcaccg tccgaagaaa agcgcggcct aaagcgcgag tctggtgact tggcacccac 14220cgtgcagctg atggtaccca agcgccagcg actggaagat gtcttggaaa aaatgaccgt 14280ggaacctggg ctggagcccg aggtccgcgt gcggccaatc aagcaggtgg cgccgggact 14340gggcgtgcag accgtggacg ttcagatacc cactaccagt agcaccagta ttgccaccgc 14400cacagagggc atggagacac aaacgtcccc ggttgcctca gcggtggcgg atgccgcggt 14460gcaggcggtc gctgcggccg cgtccaagac ctctacggag gtgcaaacgg acccgtggat 14520gtttcgcgtt tcagcccccc ggcgcccgcg ccgttcgagg aagtacggcg ccgccagcgc 14580gctactgccc gaatatgccc tacatccttc cattgcgcct acccccggct atcgtggcta 14640cacctaccgc cccagaagac gagcaactac ccgacgccga accaccactg gaacccgccg 14700ccgccgtcgc cgtcgccagc ccgtgctggc cccgatttcc gtgcgcaggg tggctcgcga 14760aggaggcagg accctggtgc tgccaacagc gcgctaccac cccagcatcg tttaaaagcc 14820ggtctttgtg gttcttgcag atatggccct cacctgccgc ctccgtttcc cggtgccggg 14880attccgagga agaatgcacc gtaggagggg catggccggc cacggcctga cgggcggcat 14940gcgtcgtgcg caccaccggc ggcggcgcgc gtcgcaccgt cgcatgcgcg gcggtatcct 15000gcccctcctt attccactga tcgccgcggc gattggcgcc gtgcccggaa ttgcatccgt 15060ggccttgcag gcgcagagac actgattaaa aacaagttgc atgtggaaaa atcaaaataa 15120aaagtctgga ctctcacgct cgcttggtcc tgtaactatt ttgtagaatg gaagacatca 15180actttgcgtc tctggccccg cgacacggct cgcgcccgtt catgggaaac tggcaagata 15240tcggcaccag caatatgagc ggtggcgcct tcagctgggg ctcgctgtgg agcggcatta 15300aaaatttcgg ttccaccgtt aagaactatg gcagcaaggc ctggaacagc agcacaggcc 15360agatgctgag ggataagttg aaagagcaaa atttccaaca aaaggtggta gatggcctgg 15420cctctggcat tagcggggtg gtggacctgg ccaaccaggc agtgcaaaat aagattaaca 15480gtaagcttga tccccgccct cccgtagagg agcctccacc ggccgtggag acagtgtctc 15540cagaggggcg tggcgaaaag cgtccgcgcc ccgacaggga agaaactctg gtgacgcaaa 15600tagacgagcc tccctcgtac gaggaggcac taaagcaagg cctgcccacc acccgtccca 15660tcgcgcccat ggctaccgga gtgctgggcc agcacacacc cgtaacgctg gacctgcctc 15720cccccgccga cacccagcag aaacctgtgc tgccaggccc gaccgccgtt gttgtaaccc 15780gtcctagccg cgcgtccctg cgccgcgccg ccagcggtcc gcgatcgttg cggcccgtag 15840ccagtggcaa ctggcaaagc acactgaaca gcatcgtggg tctgggggtg caatccctga 15900agcgccgacg atgcttctga tagctaacgt gtcgtatgtg tgtcatgtat gcgtccatgt 15960cgccgccaga ggagctgctg agccgccgcg cgcccgcttt ccaagatggc taccccttcg 16020atgatgccgc agtggtctta catgcacatc tcgggccagg acgcctcgga gtacctgagc 16080cccgggctgg tgcagtttgc ccgcgccacc gagacgtact tcagcctgaa taacaagttt 16140agaaacccca cggtggcgcc tacgcacgac gtgaccacag accggtccca gcgtttgacg 16200ctgcggttca tccctgtgga ccgtgaggat actgcgtact cgtacaaggc gcggttcacc 16260ctagctgtgg gtgataaccg tgtgctggac atggcttcca cgtactttga catccgcggc 16320gtgctggaca ggggccctac ttttaagccc tactctggca ctgcctacaa cgccctggct 16380cccaagggtg ccccaaatcc ttgcgaatgg gatgaagctg ctactgctct tgaaataaac 16440ctagaagaag aggacgatga caacgaagac gaagtagacg agcaagctga gcagcaaaaa 16500actcacgtat ttgggcaggc gccttattct ggtataaata ttacaaagga gggtattcaa 16560ataggtgtcg aaggtcaaac acctaaatat gccgataaaa catttcaacc tgaacctcaa 16620ataggagaat ctcagtggta cgaaacagaa attaatcatg cagctgggag agtcctaaaa 16680aagactaccc caatgaaacc atgttacggt tcatatgcaa aacccacaaa tgaaaatgga 16740gggcaaggca ttcttgtaaa gcaacaaaat ggaaagctag aaagtcaagt ggaaatgcaa 16800tttttctcaa ctactgaggc agccgcaggc aatggtgata acttgactcc taaagtggta 16860ttgtacagtg aagatgtaga tatagaaacc ccagacactc atatttctta catgcccact 16920attaaggaag gtaactcacg agaactaatg ggccaacaat ctatgcccaa caggcctaat 16980tacattgctt ttagggacaa ttttattggt ctaatgtatt acaacagcac gggtaatatg 17040ggtgttctgg cgggccaagc atcgcagttg aatgctgttg tagatttgca agacagaaac 17100acagagcttt cataccagct tttgcttgat tccattggtg atagaaccag gtacttttct 17160atgtggaatc aggctgttga cagctatgat ccagatgtta gaattattga aaatcatgga 17220actgaagatg aacttccaaa ttactgcttt ccactgggag gtgtgattaa tacagagact 17280cttaccaagg taaaacctaa aacaggtcag gaaaatggat gggaaaaaga tgctacagaa 17340ttttcagata aaaatgaaat aagagttgga aataattttg ccatggaaat caatctaaat 17400gccaacctgt ggagaaattt cctgtactcc aacatagcgc tgtatttgcc cgacaagcta 17460aagtacagtc cttccaacgt aaaaatttct gataacccaa acacctacga ctacatgaac 17520aagcgagtgg tggctcccgg gctagtggac tgctacatta accttggagc acgctggtcc 17580cttgactata tggacaacgt caacccattt aaccaccacc gcaatgctgg cctgcgctac 17640cgctcaatgt tgctgggcaa tggtcgctat gtgcccttcc acatccaggt gcctcagaag 17700ttctttgcca ttaaaaacct ccttctcctg ccgggctcat acacctacga gtggaacttc 17760aggaaggatg ttaacatggt tctgcagagc tccctaggaa atgacctaag ggttgacgga 17820gccagcatta agtttgatag catttgcctt tacgccacct tcttccccat ggcccacaac 17880accgcctcca cgcttgaggc catgcttaga aacgacacca acgaccagtc ctttaacgac 17940tatctctccg ccgccaacat gctctaccct atacccgcca acgctaccaa cgtgcccata 18000tccatcccct cccgcaactg ggcggctttc cgcggctggg ccttcacgcg ccttaagact 18060aaggaaaccc catcactggg ctcgggctac gacccttatt acacctactc tggctctata 18120ccctacctag atggaacctt ttacctcaac cacaccttta agaaggtggc cattaccttt 18180gactcttctg tcagctggcc tggcaatgac cgcctgctta cccccaacga gtttgaaatt 18240aagcgctcag ttgacgggga gggttacaac gttgcccagt gtaacatgac caaagactgg 18300ttcctggtac aaatgctagc taactataac attggctacc agggcttcta tatcccagag 18360agctacaagg accgcatgta ctccttcttt agaaacttcc agcccatgag ccgtcaggtg 18420gtggatgata ctaaatacaa ggactaccaa caggtgggca tcctacacca acacaacaac 18480tctggatttg ttggctacct tgcccccacc atgcgcgaag gacaggccta ccctgctaac 18540ttcccctatc cgcttatagg caagaccgca gttgacagca ttacccagaa aaagtttctt 18600tgcgatcgca ccctttggcg catcccattc tccagtaact ttatgtccat gggcgcactc 18660acagacctgg gccaaaacct tctctacgcc aactccgccc acgcgctaga catgactttt 18720gaggtggatc ccatggacga gcccaccctt ctttatgttt tgtttgaagt ctttgacgtg 18780gtccgtgtgc accagccgca ccgcggcgtc atcgaaaccg tgtacctgcg cacgcccttc 18840tcggccggca acgccacaac ataaagaagc aagcaacatc aacaacagct gccgccatgg 18900gctccagtga gcaggaactg aaagccattg tcaaagatct tggttgtggg ccatattttt 18960tgggcaccta tgacaagcgc tttccaggct ttgtttctcc acacaagctc gcctgcgcca 19020tagtcaatac ggccggtcgc gagactgggg gcgtacactg gatggccttt gcctggaacc 19080cgcactcaaa aacatgctac ctctttgagc cctttggctt ttctgaccag cgactcaagc 19140aggtttacca gtttgagtac gagtcactcc tgcgccgtag cgccattgct tcttcccccg 19200accgctgtat aacgctggaa aagtccaccc aaagcgtaca ggggcccaac tcggccgcct 19260gtggactatt ctgctgcatg tttctccacg cctttgccaa ctggccccaa actcccatgg 19320atcacaaccc caccatgaac cttattaccg gggtacccaa ctccatgctc aacagtcccc 19380aggtacagcc caccctgcgt cgcaaccagg aacagctcta cagcttcctg gagcgccact 19440cgccctactt ccgcagccac agtgcgcaga ttaggagcgc cacttctttt tgtcacttga 19500aaaacatgta aaaataatgt actagagaca ctttcaataa aggcaaatgc ttttatttgt 19560acactctcgg gtgattattt acccccaccc ttgccgtctg cgccgtttaa aaatcaaagg 19620ggttctgccg cgcatcgcta tgcgccactg gcagggacac gttgcgatac tggtgtttag 19680tgctccactt aaactcaggc acaaccatcc gcggcagctc ggtgaagttt tcactccaca 19740ggctgcgcac catcaccaac gcgtttagca ggtcgggcgc cgatatcttg aagtcgcagt 19800tggggcctcc gccctgcgcg cgcgagttgc gatacacagg gttgcagcac tggaacacta 19860tcagcgccgg gtggtgcacg ctggccagca cgctcttgtc ggagatcaga tccgcgtcca 19920ggtcctccgc gttgctcagg gcgaacggag tcaactttgg tagctgcctt cccaaaaagg 19980gcgcgtgccc aggctttgag ttgcactcgc accgtagtgg catcaaaagg tgaccgtgcc 20040cggtctgggc gttaggatac agcgcctgca taaaagcctt gatctgctta aaagccacct 20100gagcctttgc gccttcagag aagaacatgc cgcaagactt gccggaaaac tgattggccg 20160gacaggccgc gtcgtgcacg cagcaccttg cgtcggtgtt ggagatctgc accacatttc 20220ggccccaccg gttcttcacg atcttggcct tgctagactg ctccttcagc gcgcgctgcc 20280cgttttcgct cgtcacatcc atttcaatca cgtgctcctt atttatcata atgcttccgt 20340gtagacactt aagctcgcct tcgatctcag cgcagcggtg cagccacaac gcgcagcccg 20400tgggctcgtg atgcttgtag gtcacctctg caaacgactg caggtacgcc tgcaggaatc 20460gccccatcat cgtcacaaag gtcttgttgc tggtgaaggt cagctgcaac ccgcggtgct 20520cctcgttcag ccaggtcttg catacggccg ccagagcttc cacttggtca ggcagtagtt 20580tgaagttcgc ctttagatcg ttatccacgt ggtacttgtc catcagcgcg cgcgcagcct 20640ccatgccctt ctcccacgca gacacgatcg gcacactcag cgggttcatc accgtaattt 20700cactttccgc ttcgctgggc tcttcctctt cctcttgcgt ccgcatacca cgcgccactg 20760ggtcgtcttc attcagccgc cgcactgtgc gcttacctcc tttgccatgc ttgattagca 20820ccggtgggtt gctgaaaccc accatttgta gcgccacatc ttctctttct tcctcgctgt 20880ccacgattac ctctggtgat ggcgggcgct cgggcttggg agaagggcgc ttctttttct 20940tcttgggcgc aatggccaaa tccgccgccg aggtcgatgg ccgcgggctg ggtgtgcgcg 21000gcaccagcgc gtcttgtgat gagtcttcct cgtcctcgga ctcgatacgc cgcctcatcc 21060gcttttttgg gggcgcccgg ggaggcggcg gcgacgggga cggggacgac acgtcctcca 21120tggttggggg acgtcgcgcc gcaccgcgtc cgcgctcggg ggtggtttcg cgctgctcct 21180cttcccgact ggccatttcc ttctcctata ggcagaaaaa gatcatggag tcagtcgaga 21240agaaggacag cctaaccgcc ccctctgagt tcgccaccac cgcctccacc gatgccgcca 21300acgcgcctac caccttcccc gtcgaggcac ccccgcttga ggaggaggaa gtgattatcg 21360agcaggaccc aggttttgta agcgaagacg acgaggaccg ctcagtacca acagaggata 21420aaaagcaaga ccaggacaac gcagaggcaa acgaggaaca agtcgggcgg ggggacgaaa 21480ggcatggcga ctacctagat gtgggagacg acgtgctgtt gaagcatctg cagcgccagt 21540gcgccattat ctgcgacgcg ttgcaagagc gcagcgatgt gcccctcgcc atagcggatg 21600tcagccttgc ctacgaacgc cacctattct caccgcgcgt accccccaaa cgccaagaaa 21660acggcacatg cgagcccaac ccgcgcctca acttctaccc cgtatttgcc gtgccagagg 21720tgcttgccac ctatcacatc tttttccaaa actgcaagat acccctatcc tgccgtgcca 21780accgcagccg agcggacaag cagctggcct tgcggcaggg cgctgtcata cctgatatcg 21840cctcgctcaa cgaagtgcca aaaatctttg agggtcttgg acgcgacgag aagcgcgcgg 21900caaacgctct gcaacaggaa aacagcgaaa atgaaagtca ctctggagtg ttggtggaac 21960tcgagggtga caacgcgcgc ctagccgtac taaaacgcag catcgaggtc acccactttg 22020cctacccggc acttaaccta ccccccaagg tcatgagcac agtcatgagt gagctgatcg 22080tgcgccgtgc gcagcccctg gagagggatg caaatttgca agaacaaaca gaggagggcc 22140tacccgcagt tggcgacgag cagctagcgc gctggcttca aacgcgcgag cctgccgact 22200tggaggagcg acgcaaacta atgatggccg cagtgctcgt taccgtggag cttgagtgca 22260tgcagcggtt ctttgctgac ccggagatgc agcgcaagct agaggaaaca ttgcactaca 22320cctttcgaca gggctacgta cgccaggcct gcaagatctc caacgtggag ctctgcaacc 22380tggtctccta ccttggaatt ttgcacgaaa accgccttgg gcaaaacgtg cttcattcca 22440cgctcaaggg cgaggcgcgc cgcgactacg tccgcgactg cgtttactta tttctatgct 22500acacctggca gacggccatg ggcgtttggc agcagtgctt ggaggagtgc aacctcaagg 22560agctgcagaa actgctaaag caaaacttga aggacctatg gacggccttc aacgagcgct 22620ccgtggccgc gcacctggcg gacatcattt tccccgaacg cctgcttaaa accctgcaac 22680agggtctgcc agacttcacc agtcaaagca tgttgcagaa ctttaggaac tttatcctag 22740agcgctcagg aatcttgccc gccacctgct gtgcacttcc tagcgacttt gtgcccatta 22800agtaccgcga atgccctccg ccgctttggg gccactgcta ccttctgcag ctagccaact 22860accttgccta ccactctgac ataatggaag acgtgagcgg tgacggtcta ctggagtgtc 22920actgtcgctg caacctatgc accccgcacc gctccctggt ttgcaattcg cagctgctta 22980acgaaagtca aattatcggt acctttgagc tgcagggtcc ctcgcctgac gaaaagtccg 23040cggctccggg gttgaaactc actccggggc tgtggacgtc ggcttacctt cgcaaatttg 23100tacctgagga ctaccacgcc cacgagatta ggttctacga agaccaatcc cgcccgccta 23160atgcggagct taccgcctgc gtcattaccc agggccacat tcttggccaa ttgcaagcca 23220tcaacaaagc ccgccaagag tttctgctac gaaagggacg gggggtttac ttggaccccc 23280agtccggcga ggagctcaac ccaatccccc cgccgccgca gccctatcag cagcagccgc 23340gggcccttgc ttcccaggat ggcacccaaa aagaagctgc agctgccgcc gccacccacg 23400gacgaggagg aatactggga cagtcaggca gaggaggttt tggacgagga ggaggaggac 23460atgatggaag actgggagag cctagacgag gaagcttccg aggtcgaaga ggtgtcagac 23520gaaacaccgt caccctcggt cgcattcccc tcgccggcgc cccagaaatc ggcaaccggt 23580tccagcatgg ctacaacctc cgctcctcag gcgccgccgg cactgcccgt tcgccgaccc 23640aaccgtagat gggacaccac tggaaccagg gccggtaagt ccaagcagcc gccgccgtta 23700gcccaagagc aacaacagcg ccaaggctac cgctcatggc gcgggcacaa gaacgccata 23760gttgcttgct tgcaagactg tgggggcaac atctccttcg cccgccgctt tcttctctac 23820catcacggcg tggccttccc ccgtaacatc ctgcattact accgtcatct ctacagccca 23880tactgcaccg gcggcagcgg cagcaacagc agcggccaca cagaagcaaa ggcgaccgga 23940tagcaagact ctgacaaagc ccaagaaatc cacagcggcg gcagcagcag gaggaggagc 24000gctgcgtctg gcgcccaacg aacccgtatc gacccgcgag cttagaaaca ggatttttcc 24060cactctgtat gctatatttc aacagagcag gggccaagaa caagagctga aaataaaaaa 24120caggtctctg cgatccctca cccgcagctg cctgtatcac aaaagcgaag atcagcttcg 24180gcgcacgctg gaagacgcgg aggctctctt cagtaaatac tgcgcgctga ctcttaagga 24240ctagtttcgc gccctttctc aaatttaagc gcgaaaacta cgtcatctcc agcggccaca 24300cccggcgcca gcacctgttg tcagcgccat tatgagcaag gaaattccca cgccctacat 24360gtggagttac cagccacaaa tgggacttgc ggctggagct gcccaagact actcaacccg 24420aataaactac atgagcgcgg gaccccacat gatatcccgg gtcaacggaa tacgcgccca 24480ccgaaaccga attctcctgg aacaggcggc tattaccacc acacctcgta ataaccttaa 24540tccccgtagt tggcccgctg ccctggtgta ccaggaaagt cccgctccca ccactgtggt 24600acttcccaga gacgcccagg ccgaagttca gatgactaac tcaggggcgc agcttgcggg 24660cggctttcgt cacagggtgc ggtcgcccgg gcagggtata actcacctga caatcagagg 24720gcgaggtatt cagctcaacg acgagtcggt gagctcctcg cttggtctcc gtccggacgg 24780gacatttcag atcggcggcg ccggccgctc ttcattcacg cctcgtcagg caatcctaac 24840tctgcagacc tcgtcctctg agccgcgctc tggaggcatt ggaactctgc aatttattga 24900ggagtttgtg ccatcggtct actttaaccc cttctcggga cctcccggcc actatccgga 24960tcaatttatt cctaactttg acgcggtaaa ggactcggcg gacggctacg actgaatgtt 25020aagtggagag gcagagcaac tgcgcctgaa acacctggtc cactgtcgcc gccacaagtg 25080ctttgcccgc gactccggtg agttttgcta ctttgaattg cccgaggatc atatcgaggg 25140cccggcgcac ggcgtccggc ttaccgccca gggagagctt gcccgtagcc tgattcggga 25200gtttacccag cgccccctgc tagttgagcg ggacagggga ccctgtgttc tcactgtgat 25260ttgcaactgt cctaaccctg gattacatca agatctttgt tgccatctct gtgctgagta 25320taataaatac agaaattaaa atatactggg gctcctatcg ccatcctgta aacgccaccg 25380tcttcacccg cccaagcaaa ccaaggcgaa ccttacctgg tacttttaac atctctccct 25440ctgtgattta caacagtttc aacccagacg gagtgagtct acgagagaac ctctccgagc 25500tcagctactc catcagaaaa aacaccaccc tccttacctg ccgggaacgt acgagtgcgt 25560caccggccgc tgcaccacac ctaccgcctg accgtaaacc agactttttc cggacagacc 25620tcaataactc tgtttaccag aacaggaggt gagcttagaa aacccttagg gtattaggcc 25680aaaggcgcag ctactgtggg gtttatgaac aattcaagca actctacggg ctattctaat 25740tcaggtttct ctagaatcgg ggttggggtt attctctgtc ttgtgattct ctttattctt 25800atactaacgc ttctctgcct aaggctcgcc gcctgctgtg tgcacatttg catttattgt 25860cagcttttta aacgctgggg tcgccaccca agatgattag gtacataatc ctaggtttac 25920tcacccttgc gtcagcccac ggtaccaccc aaaaggtgga ttttaaggag ccagcctgta 25980atgttacatt cgcagctgaa gctaatgagt gcaccactct tataaaatgc accacagaac 26040atgaaaagct gcttattcgc cacaaaaaca aaattggcaa gtatgctgtt

tatgctattt 26100ggcagccagg tgacactaca gagtataatg ttacagtttt ccagggtaaa agtcataaaa 26160cttttatgta tacttttcca ttttatgaaa tgtgcgacat taccatgtac atgagcaaac 26220agtataagtt gtggccccca caaaattgtg tggaaaacac tggcactttc tgctgcactg 26280ctatgctaat tacagtgctc gctttggtct gtaccctact ctatattaaa tacaaaagca 26340gacgcagctt tattgaggaa aagaaaatgc cttaatttac taagttacaa agctaatgtc 26400accactaact gctttactcg ctgcttgcaa aacaaattca aaaagttagc attataatta 26460gaataggatt taaacccccc ggtcatttcc tgctcaatac cattcccctg aacaattgac 26520tctatgtggg atatgctcca gcgctacaac cttgaagtca ggcttcctgg atgtcagcat 26580ctgactttgg ccagcacctg tcccgcggat ttgttccagt ccaactacag cgacccaccc 26640taacagagat gaccaacaca accaacgcgg ccgccgctac cggacttaca tctaccacaa 26700atacacccca agtttctgcc tttgtcaata actgggataa cttgggcatg tggtggttct 26760ccatagcgct tatgtttgta tgccttatta ttatgtggct catctgctgc ctaaagcgca 26820aacgcgcccg accacccatc tatagtccca tcattgtgct acacccaaac aatgatggaa 26880tccatagatt ggacggactg aaacacatgt tcttttctct tacagtatga ttaaatgaga 26940catgattcct cgagttttta tattactgac ccttgttgcg ctttttttgt gcgtgctcca 27000cattggctgc ggtttctcac atcgaagtag actgcattcc agccttcaca gtctatttgc 27060tttacggatt tgtcaccctc acgctcatct gcagcctcat cactgtggtc atcgccttta 27120tccagtgcat tgactgggtc tgtgtgcgct ttgcatatct cagacaccat ccccagtaca 27180gggacaggac tatagctgag cttcttagaa ttctttaatt atgaaattta ctgtgacttt 27240tctgctgatt atttgcaccc tatctgcgtt ttgttccccg acctccaagc ctcaaagaca 27300tatatcatgc agattcactc gtatatggaa tattccaagt tgctacaatg aaaaaagcga 27360tctttccgaa gcctggttat atgcaatcat ctctgttatg gtgttctgca gtaccatctt 27420agccctagct atatatccct accttgacat tggctggaac gcaatagatg ccatgaacca 27480cccaactttc cccgcgcccg ctatgcttcc actgcaacaa gttgttgccg gcggctttgt 27540cccagccaat cagcctcgcc caccttctcc cacccccact gaaatcagct actttaatct 27600aacaggagga gatgactgac accctagatc tagaaatgga cggaattatt acagagcagc 27660gcctgctaga aagacgcagg gcagcggccg agcaacagcg catgaatcaa gagctccaag 27720acatggttaa cttgcaccag tgcaaaaggg gtatcttttg tctggtaaag caggccaaag 27780tcacctacga cagtaatacc accggacacc gccttagcta caagttgcca accaagcgtc 27840agaaattggt ggtcatggtg ggagaaaagc ccattaccat aactcagcac tcggtagaaa 27900ccgaaggctg cattcactca ccttgtcaag gacctgagga tctctgcacc cttattaaga 27960ccctgtgcgg tctcaaagat cttattccct ttaactaata aaaaaaaata ataaagcatc 28020acttacttaa aatcagttag caaatttctg tccagtttat tcagcagcac ctccttgccc 28080tcctcccagc tctggtattg cagcttcctc ctggctgcaa actttctcca caatctaaat 28140ggaatgtcag tttcctcctg ttcctgtcca tccgcaccca ctatcttcat gttgttgcag 28200atgaagcgcg caagaccgtc tgaagatacc ttcaaccccg tgtatccata tgacacggaa 28260accggtcctc caactgtgcc ttttcttact cctccctttg tatcccccaa tgggtttcaa 28320gagagtcccc ctggggtact ctctttgcgc ctatccgaac ctctagttac ctccaatggc 28380atgcttgcgc tcaaaatggg caacggcctc tctctggacg aggccggcaa ccttacctcc 28440caaaatgtaa ccactgtgag cccacctctc aaaaaaacca agtcaaacat aaacctggaa 28500atatctgcac ccctcacagt tacctcagaa gccctaactg tggctgccgc cgcacctcta 28560atggtcgcgg gcaacacact caccatgcaa tcacaggccc cgctaaccgt gcacgactcc 28620aaacttagca ttgccaccca aggacccctc acagtgtcag aaggaaagct agccctgcaa 28680acatcaggcc ccctcaccac caccgatagc agtaccctta ctatcactgc ctcaccccct 28740ctaactactg ccactggtag cttgggcatt gacttgaaag agcccattta tacacaaaat 28800ggaaaactag gactaaagta cggggctcct ttgcatgtaa cagacgacct aaacactttg 28860accgtagcaa ctggtccagg tgtgactatt aataatactt ccttgcaaac taaagttact 28920ggagccttgg gttttgattc acaaggcaat atgcaactta atgtagcagg aggactaagg 28980attgattctc aaaacagacg ccttatactt gatgttagtt atccgtttga tgctcaaaac 29040caactaaatc taagactagg acagggccct ctttttataa actcagccca caacttggat 29100attaactaca acaaaggcct ttacttgttt acagcttcaa acaattccaa aaagcttgag 29160gttaacctaa gcactgccaa ggggttgatg tttgacgcta cagccatagc cattaatgca 29220ggagatgggc ttgaatttgg ttcacctaat gcaccaaaca caaatcccct caaaacaaaa 29280attggccatg gcctagaatt tgattcaaac aaggctatgg ttcctaaact aggaactggc 29340cttagttttg acagcacagg tgccattaca gtaggaaaca aaaataatga taagctaact 29400ttgtggacca caccagctcc atctcctaac tgtagactaa atgcagagaa agatgctaaa 29460ctcactttgg tcttaacaaa atgtggcagt caaatacttg ctacagtttc agttttggct 29520gttaaaggca gtttggctcc aatatctgga acagttcaaa gtgctcatct tattataaga 29580tttgacgaaa atggagtgct actaaacaat tccttcctgg acccagaata ttggaacttt 29640agaaatggag atcttactga aggcacagcc tatacaaacg ctgttggatt tatgcctaac 29700ctatcagctt atccaaaatc tcacggtaaa actgccaaaa gtaacattgt cagtcaagtt 29760tacttaaacg gagacaaaac taaacctgta acactaacca ttacactaaa cggtacacag 29820gaaacaggag acacaactcc aagtgcatac tctatgtcat tttcatggga ctggtctggc 29880cacaactaca ttaatgaaat atttgccaca tcctcttaca ctttttcata cattgcccaa 29940gaataaagaa tcgtttgtgt tatgtttcaa cgtgtttatt tttcaattgc agaaaatttc 30000aagtcatttt tcattcagta gtatagcccc accaccacat agcttataca gatcaccgta 30060ccttaatcaa actcacagaa ccctagtatt caacctgcca cctccctccc aacacacaga 30120gtacacagtc ctttctcccc ggctggcctt aaaaagcatc atatcatggg taacagacat 30180attcttaggt gttatattcc acacggtttc ctgtcgagcc aaacgctcat cagtgatatt 30240aataaactcc ccgggcagct cacttaagtt catgtcgctg tccagctgct gagccacagg 30300ctgctgtcca acttgcggtt gcttaacggg cggcgaagga gaagtccacg cctacatggg 30360ggtagagtca taatcgtgca tcaggatagg gcggtggtgc tgcagcagcg cgcgaataaa 30420ctgctgccgc cgccgctccg tcctgcagga atacaacatg gcagtggtct cctcagcgat 30480gattcgcacc gcccgcagca taaggcgcct tgtcctccgg gcacagcagc gcaccctgat 30540ctcacttaaa tcagcacagt aactgcagca cagcaccaca atattgttca aaatcccaca 30600gtgcaaggcg ctgtatccaa agctcatggc ggggaccaca gaacccacgt ggccatcata 30660ccacaagcgc aggtagatta agtggcgacc cctcataaac acgctggaca taaacattac 30720ctcttttggc atgttgtaat tcaccacctc ccggtaccat ataaacctct gattaaacat 30780ggcgccatcc accaccatcc taaaccagct ggccaaaacc tgcccgccgg ctatacactg 30840cagggaaccg ggactggaac aatgacagtg gagagcccag gactcgtaac catggatcat 30900catgctcgtc atgatatcaa tgttggcaca acacaggcac acgtgcatac acttcctcag 30960gattacaagc tcctcccgcg ttagaaccat atcccaggga acaacccatt cctgaatcag 31020cgtaaatccc acactgcagg gaagacctcg cacgtaactc acgttgtgca ttgtcaaagt 31080gttacattcg ggcagcagcg gatgatcctc cagtatggta gcgcgggttt ctgtctcaaa 31140aggaggtaga cgatccctac tgtacggagt gcgccgagac aaccgagatc gtgttggtcg 31200tagtgtcatg ccaaatggaa cgccggacgt agtcatattt cctgaagcaa aaccaggtgc 31260gggcgtgaca aacagatctg cgtctccggt ctcgccgctt agatcgctct gtgtagtagt 31320tgtagtatat ccactctctc aaagcatcca ggcgccccct ggcttcgggt tctatgtaaa 31380ctccttcatg cgccgctgcc ctgataacat ccaccaccgc agaataagcc acacccagcc 31440aacctacaca ttcgttctgc gagtcacaca cgggaggagc gggaagagct ggaagaacca 31500tgtttttttt tttattccaa aagattatcc aaaacctcaa aatgaagatc tattaagtga 31560acgcgctccc ctccggtggc gtggtcaaac tctacagcca aagaacagat aatggcattt 31620gtaagatgtt gcacaatggc ttccaaaagg caaacggccc tcacgtccaa gtggacgtaa 31680aggctaaacc cttcagggtg aatctcctct ataaacattc cagcaccttc aaccatgccc 31740aaataattct catctcgcca ccttctcaat atatctctaa gcaaatcccg aatattaagt 31800ccggccattg taaaaatctg ctccagagcg ccctccacct tcagcctcaa gcagcgaatc 31860atgattgcaa aaattcaggt tcctcacaga cctgtataag attcaaaagc ggaacattaa 31920caaaaatacc gcgatcccgt aggtcccttc gcagggccag ctgaacataa tcgtgcaggt 31980ctgcacggac cagcgcggcc acttccccgc caggaaccat gacaaaagaa cccacactga 32040ttatgacacg catactcgga gctatgctaa ccagcgtagc cccgatgtaa gcttgttgca 32100tgggcggcga tataaaatgc aaggtgctgc tcaaaaaatc aggcaaagcc tcgcgcaaaa 32160aagaaagcac atcgtagtca tgctcatgca gataaaggca ggtaagctcc ggaaccacca 32220cagaaaaaga caccattttt ctctcaaaca tgtctgcggg tttctgcata aacacaaaat 32280aaaataacaa aaaaacattt aaacattaga agcctgtctt acaacaggaa aaacaaccct 32340tataagcata agacggacta cggccatgcc ggcgtgaccg taaaaaaact ggtcaccgtg 32400attaaaaagc accaccgaca gctcctcggt catgtccgga gtcataatgt aagactcggt 32460aaacacatca ggttgattca catcggtcag tgctaaaaag cgaccgaaat agcccggggg 32520aatacatacc cgcaggcgta gagacaacat tacagccccc ataggaggta taacaaaatt 32580aataggagag aaaaacacat aaacacctga aaaaccctcc tgcctaggca aaatagcacc 32640ctcccgctcc agaacaacat acagcgcttc cacagcggca gccataacag tcagccttac 32700cagtaaaaaa gaaaacctat taaaaaaaca ccactcgaca cggcaccagc tcaatcagtc 32760acagtgtaaa aaagggccaa gtgcagagcg agtatatata ggactaaaaa atgacgtaac 32820ggttaaagtc cacaaaaaac acccagaaaa ccgcacgcga acctacgccc agaaacgaaa 32880gccaaaaaac ccacaacttc ctcaaatcgt cacttccgtt ttcccacgtt acgtcacttc 32940ccattttaag aaaactacaa ttcccaacac atacaagtta ctccgcccta aaacctacgt 33000cacccgcccc gttcccacgc cccgcgccac gtcacaaact ccaccccctc attatcatat 33060tggcttcaat ccaaaataag gtatattatt gat 3309312978DNAArtificial sequencePPE-1-3X promoter 12acgtgtactt ctgatcggcg atactaggga gataaggatg tacctgacaa aaccacattg 60ttgttgttat cattattatt tagttttcct tccttgctaa ctcctgacgg aatctttctc 120acctcaaatg cgaagtactt tagtttagaa aagacttggt ggaaggggtg gtggtggaaa 180agtagggtga tcttccaaac taatctggtt ccccgcccgc cccagtagct gggattcaag 240agcgaagagt ggggatcgtc cccttgtttg atcagaaaga cataaaagga aaatcaagtg 300aacaatgatc agccccacct ccaccccacc cccctgcgcg cgcacaatac aatctattta 360attgtacttc atacttttca ttccaatggg gtgactttgc ttctggagaa actcttgatt 420cttgaactct ggggctggca gctagcctcc agaagcaaag tcaccccatt ggaatgaaaa 480gtatgaagta caatgaaaag tatgaagtac tggctccaga agcaaagtca ccctccagaa 540gcaaagtcac cccattggaa tgaaaagtat gaagtacgct agcaaaaggg gaagcgggct 600gctgctctct gcaggttctg cagcggtctc tgtctagtgg gtgttttctt tttcttagcc 660ctgcccctgg attgtcagac ggcgggcgtc tgcctctgaa gttagccgtg atttcctcta 720gagccgggtc ttatctctgg ctgcacgttg cctgtgggtg actaatcaca caataacatt 780gtttagggct ggaataaagt cagagctgtt tacccccact ctataggggt tcaatataaa 840aaggcggcgg agaactgtcc gagtcagaag cgttcctgca ccggcgctga gagcctgacc 900cggtctgctc cgctgtcctt gcgcgctgcc tcccggctgc ccgcgacgct ttcgccccag 960tggaagggcc acttgctg 978131334DNAMus musculus 13gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt gtgtagtgta cttctgatcg 60gcgatactag ggagataagg atgtacctga caaaaccaca ttgttgttgt tatcattatt 120atttagtttt ccttccttgc taactcctga cggaatcttt ctcacctcaa atgcgaagta 180ctttagttta gaaaagactt ggtggaaggg gtggtggtgg aaaagtaggg tgatcttcca 240aactaatctg gttccccgcc cgccccagta gctgggattc aagagcgaag agtggggatc 300gtccccttgt ttgatcagaa agacataaaa ggaaaatcaa gtgaacaatg atcagcccca 360cctccacccc acccccctgc gcgcgcacaa tacaatctat ttaattgtac ttcatacttt 420tcattccaat ggggtgactt tgcttctgga gaaactcttg attcttgaac tctggggctg 480gcagctagca aaaggggaag cgggctgctg ctctctgcag gttctgcagc ggtctctgtc 540tagtgggtgt tttctttttc ttagccctgc ccctggattg tcagacggcg ggcgtctgcc 600tctgaagtta gccgtgattt cctctagagc cgggtcttat ctctggctgc acgttgcctg 660tgggtgacta atcacacaat aacattgttt agggctggaa taaagtcaga gctgtttacc 720cccactctat aggggttcaa tataaaaagg cggcggagaa ctgtccgagt cagacgcgtt 780cctgcaccgg cgctgagagc ctgacccggt ctgctccgct gtccttgcgc gctgcctccc 840ggctgcccgc gacgctttcg ccccagtgga agggccactt gctgaggacc gcgctgagat 900ctaaaaaaaa aacaaaaaac aaaaaacaaa aaaacccaga ggcgatcaga gcgaccagac 960accgtcctct tcgttttgca ttgagttcca tttgcaaccg agttttcttt ttttcctttt 1020tccccactct tctgacccct ttgcagaatg gattattttc ccgtgatctt ctctctgctg 1080ttcgtgactt tccaaggagc tccagaaaca ggtaggcgcc acttgcgaat ctttctactt 1140cagcgcagca gttatcgctt ctgttttcca cttttctttc tttcttttct ttcattcttt 1200cctttttatt tattttttta attactgaag ctccagcagc aagtgcctta caattaatta 1260acttctgtgt gaagcgaaag aaataaaacc cctgtttgaa tacagctgac tacaaccgag 1320tatcgcatag cttc 1334145DNAArtificial sequencewild type element M9 14ctgga 5155DNAArtificial sequencewild type element M4 15cattc 5165DNAArtificial sequencewild type element M5 16caatg 5175DNAArtificial sequencewild type element M6 17gggtg 5185DNAArtificial sequencewild type element M7 18acttt 5195DNAArtificial sequencewild type element M8 19gcttc 5205DNAArtificial sequencewild type element M1 20gtact 5215DNAArtificial sequencewild type element M3 21ctttt 52219DNAArtificial sequenceExemplary element X variant 22gggtgacttt gcttctgga 192319DNAArtificial sequenceExemplary element X variant 23gggtgacttt gcttctgga 192419DNAArtificial sequenceExemplary element X variant 24ggggtgactt tgcttctgg 192519DNAArtificial sequenceExemplary element X variant 25caatggggtg gcttctgga 192619DNAArtificial sequenceExemplary element X variant 26ccaatggggt ggcttctgg 192719DNAArtificial sequenceExemplary element X variant 27gggtgacttt gcttctgga 192819DNAArtificial sequenceExemplary element X variant 28ggggtgactt tgcttctgg 192944DNAArtificial sequenceExemplary element X variant 29gtacttcata cttttcattc caatggggtg actttgcttc tgga 443039DNAArtificial sequenceExemplary element X variant 30gtactctttt cattccaatg gggtgacttt gcttctgga 393139DNAArtificial sequenceExemplary element X variant 31gtacttcata cattccaatg gggtgacttt gcttctgga 393239DNAArtificial sequenceExemplary element X variant 32gtacttcata cttttcaatg gggtgacttt gcttctgga 393339DNAArtificial sequenceExemplary element X variant 33gtacttcata cttttcattc gggtgacttt gcttctgga 393434DNAArtificial sequenceExemplary element X variant 34gtactcattc caatggggtg actttgcttc tgga 343529DNAArtificial sequenceExemplary element X variant 35gtactcaatg gggtgacttt gcttctgga 293624DNAArtificial sequenceExemplary element X variant 36gtactgggtg actttgcttc tgga 243734DNAArtificial sequenceExemplary element X variant 37gtacttcata caatggggtg actttgcttc tgga 343829DNAArtificial sequenceExemplary element X variant 38gtacttcata gggtgacttt gcttctgga 293934DNAArtificial sequenceExemplary element X variant 39gtacttcata cttttgggtg actttgcttc tgga 344034DNAArtificial sequenceExemplary element X variant 40gtactctttt caatggggtg actttgcttc tgga 344134DNAArtificial sequenceExemplary element X variant 41gtacttcata cattcgggtg actttgcttc tgga 344234DNAArtificial sequenceExemplary element X variant 42gtactctttt cattcgggtg actttgcttc tgga 344344DNAArtificial sequenceExemplary element X variant 43gtacttcata cttttcattc caatggggtg actttgcttc tgga 444439DNAArtificial sequenceExemplary element X variant 44gtactctttt cattccaatg gggtgacttt gcttctgga 394539DNAArtificial sequenceExemplary element X variant 45gtacttcata cattccaatg gggtgacttt gcttctgga 394639DNAArtificial sequenceExemplary element X variant 46gtacttcata cttttcaatg gggtgacttt gcttctgga 394739DNAArtificial sequenceExemplary element X variant 47gtacttcata cttttcattc gggtgacttt gcttctgga 394839DNAArtificial sequenceExemplary element X variant 48gtacttcata cttttcattc caatgacttt gcttctgga 394939DNAArtificial sequenceExemplary element X variant 49gtacttcata cttttcattc caatggggtg gcttctgga 395034DNAArtificial sequenceExemplary element X variant 50gtactcattc caatggggtg actttgcttc tgga 345129DNAArtificial sequenceExemplary element X variant 51gtactcaatg gggtgacttt gcttctgga 295224DNAArtificial sequenceExemplary element X variant 52gtactgggtg actttgcttc tgga 245319DNAArtificial sequenceExemplary element X variant 53gtactacttt gcttctgga 195434DNAArtificial sequenceExemplary element X variant 54gtacttcata caatggggtg actttgcttc tgga 345528DNAArtificial sequenceExemplary element X variant 55cattccaatg gggtgacttt gcttctgg 285623DNAArtificial sequenceExemplary element X variant 56cattcgggtg actttgcttc tgg 235719DNAArtificial sequenceExemplary element X variant 57cattcacttt gcttctgga 195819DNAArtificial sequenceExemplary element X variant 58cttttcattc gcttctgga 195923DNAArtificial sequenceExemplary element X variant 59cattccaatg actttgcttc tgg 236019DNAArtificial sequenceExemplary element X variant 60cattccaatg gcttctgga 196123DNAArtificial sequenceExemplary element X variant 61cattccaatg gggtggcttc tgg

236219DNAArtificial sequenceExemplary element X variant 62cattcgggtg gcttctgga 196323DNAArtificial sequenceExemplary element X variant 63caatggggtg actttgcttc tgg 236419DNAArtificial sequenceExemplary element X variant 64caatggggtg gcttctgga 196519DNAArtificial sequenceExemplary element X variant 65caatgacttt gcttctgga 196619DNAArtificial sequenceExemplary element X variant 66cattccaatg gcttctgga 196728DNAArtificial sequenceExemplary element X variant 67cattccaatg gggtgacttt gcttctgg 286823DNAArtificial sequenceExemplary element X variant 68cattccaatg actttgcttc tgg 236919DNAArtificial sequenceExemplary element X variant 69cattccaatg gcttctgga 197023DNAArtificial sequenceExemplary element X variant 70cattccaatg gggtggcttc tgg 237143DNAArtificial sequenceExemplary element X variant 71gtacttcata cttttcattc caatggggtg actttgcttc tgg 437243DNAArtificial sequenceExemplary element X variant 72gtacttcata cttttcattc caatggggtg actttgcttc tgg 437338DNAArtificial sequenceExemplary element X variant 73gtactctttt cattccaatg gggtgacttt gcttctgg 387438DNAArtificial sequenceExemplary element X variant 74gtacttcata cattccaatg gggtgacttt gcttctgg 387533DNAArtificial sequenceExemplary element X variant 75gtactcattc caatggggtg actttgcttc tgg 337643DNAArtificial sequenceExemplary element X variant 76gtacttcata cttttcattc caatggggtg actttgcttc tgg 437738DNAArtificial sequenceExemplary element X variant 77gtacttcata cttttcattc gggtgacttt gcttctgg 387838DNAArtificial sequenceExemplary element X variant 78gtacttcata cttttcattc caatgacttt gcttctgg 387938DNAArtificial sequenceExemplary element X variant 79gtacttcata cttttcattc caatggggtg gcttctgg 388033DNAArtificial sequenceExemplary element X variant 80gtacttcata cttttcattc actttgcttc tgg 338133DNAArtificial sequenceExemplary element X variant 81gtacttcata cttttcattc gggtggcttc tgg 338228DNAArtificial sequenceExemplary element X variant 82gtacttcata cttttcattc gcttctgg 288333DNAArtificial sequenceExemplary element X variant 83gtacttcata cttttcattc caatggcttc tgg 338433DNAArtificial sequenceExemplary element X variant 84gtacttcata cattcgggtg actttgcttc tgg 338533DNAArtificial sequenceExemplary element X variant 85gtacttcata cattccaatg actttgcttc tgg 338633DNAArtificial sequenceExemplary element X variant 86gtacttcata cattccaatg gggtggcttc tgg 338728DNAArtificial sequenceExemplary element X variant 87gtacttcata cattcacttt gcttctgg 288828DNAArtificial sequenceExemplary element X variant 88gtacttcata cattcgggtg gcttctgg 288923DNAArtificial sequenceExemplary element X variant 89gtacttcata cattcgcttc tgg 239028DNAArtificial sequenceExemplary element X variant 90gtacttcata cattccaatg gcttctgg 289138DNAArtificial sequenceExemplary element X variant 91gtactctttt cattccaatg gggtgacttt gcttctgg 389233DNAArtificial sequenceExemplary element X variant 92gtactctttt cattcgggtg actttgcttc tgg 339333DNAArtificial sequenceExemplary element X variant 93gtactctttt cattccaatg actttgcttc tgg 339433DNAArtificial sequenceExemplary element X variant 94gtactctttt cattccaatg gggtggcttc tgg 339528DNAArtificial sequenceExemplary element X variant 95gtactctttt cattcacttt gcttctgg 289628DNAArtificial sequenceExemplary element X variant 96gtactctttt cattcgggtg gcttctgg 289723DNAArtificial sequenceExemplary element X variant 97gtactctttt cattcgcttc tgg 239828DNAArtificial sequenceExemplary element X variant 98gtactctttt cattccaatg gcttctgg 289933DNAArtificial sequenceExemplary element X variant 99gtactcattc caatggggtg actttgcttc tgg 3310028DNAArtificial sequenceExemplary element X variant 100gtactcattc gggtgacttt gcttctgg 2810128DNAArtificial sequenceExemplary element X variant 101gtactcattc caatgacttt gcttctgg 2810228DNAArtificial sequenceExemplary element X variant 102gtactcattc caatggggtg gcttctgg 2810323DNAArtificial sequenceExemplary element X variant 103gtactcattc actttgcttc tgg 2310423DNAArtificial sequenceExemplary element X variant 104gtactcattc gggtggcttc tgg 2310523DNAArtificial sequenceExemplary element X variant 105gtactcattc caatggcttc tgg 2310643DNAArtificial sequenceExemplary element X variant 106gtacttcata cttttcattc caatggggtg actttgcttc tgg 4310738DNAArtificial sequenceExemplary element X variant 107gtactctttt cattccaatg gggtgacttt gcttctgg 3810838DNAArtificial sequenceExemplary element X variant 108gtacttcata cattccaatg gggtgacttt gcttctgg 3810938DNAArtificial sequenceExemplary element X variant 109gtacttcata cttttcaatg gggtgacttt gcttctgg 3811033DNAArtificial sequenceExemplary element X variant 110gtactcattc caatggggtg actttgcttc tgg 3311133DNAArtificial sequenceExemplary element X variant 111gtacttcata caatggggtg actttgcttc tgg 3311233DNAArtificial sequenceExemplary element X variant 112gtactctttt caatggggtg actttgcttc tgg 3311328DNAArtificial sequenceExemplary element X variant 113gtactcaatg gggtgacttt gcttctgg 2811438DNAArtificial sequenceExemplary element X variant 114gtacttcata cttttcattc caatgacttt gcttctgg 3811538DNAArtificial sequenceExemplary element X variant 115gtacttcata cttttcattc caatggggtg gcttctgg 3811633DNAArtificial sequenceExemplary element X variant 116gtacttcata cttttcattc caatggcttc tgg 3311733DNAArtificial sequenceExemplary element X variant 117gtactctttt cattccaatg actttgcttc tgg 3311833DNAArtificial sequenceExemplary element X variant 118gtacttcata cattccaatg actttgcttc tgg 3311933DNAArtificial sequenceExemplary element X variant 119gtacttcata cttttcaatg actttgcttc tgg 3312028DNAArtificial sequenceExemplary element X variant 120gtactcattc caatgacttt gcttctgg 2812128DNAArtificial sequenceExemplary element X variant 121gtacttcata caatgacttt gcttctgg 2812228DNAArtificial sequenceExemplary element X variant 122gtactctttt caatgacttt gcttctgg 2812323DNAArtificial sequenceExemplary element X variant 123gtactcaatg actttgcttc tgg 2312433DNAArtificial sequenceExemplary element X variant 124gtactctttt cattccaatg gggtggcttc tgg 3312533DNAArtificial sequenceExemplary element X variant 125gtacttcata cattccaatg gggtggcttc tgg 3312633DNAArtificial sequenceExemplary element X variant 126gtacttcata cttttcaatg gggtggcttc tgg 3312728DNAArtificial sequenceExemplary element X variant 127gtactcattc caatggggtg gcttctgg 2812828DNAArtificial sequenceExemplary element X variant 128gtacttcata caatggggtg gcttctgg 2812928DNAArtificial sequenceExemplary element X variant 129gtactctttt caatggggtg gcttctgg 2813023DNAArtificial sequenceExemplary element X variant 130gtactcaatg gggtggcttc tgg 2313128DNAArtificial sequenceExemplary element X variant 131gtactctttt cattccaatg gcttctgg 2813228DNAArtificial sequenceExemplary element X variant 132gtacttcata cattccaatg gcttctgg 2813328DNAArtificial sequenceExemplary element X variant 133gtacttcata cttttcaatg gcttctgg 2813423DNAArtificial sequenceExemplary element X variant 134gtactcattc caatggcttc tgg 2313523DNAArtificial sequenceExemplary element X variant 135gtacttcata caatggcttc tgg 2313623DNAArtificial sequenceExemplary element X variant 136gtactctttt caatggcttc tgg 2313743DNAArtificial sequenceExemplary element X variant 137gtacttcata cttttcattc caatggggtg actttgcttc tgg 4313838DNAArtificial sequenceExemplary element X variant 138gtactctttt cattccaatg gggtgacttt gcttctgg 3813938DNAArtificial sequenceExemplary element X variant 139gtacttcata cattccaatg gggtgacttt gcttctgg 3814033DNAArtificial sequenceExemplary element X variant 140gtactcattc caatggggtg actttgcttc tgg 3314138DNAArtificial sequenceExemplary element X variant 141gtacttcata cttttcattc caatgacttt gcttctgg 3814238DNAArtificial sequenceExemplary element X variant 142gtacttcata cttttcattc caatggggtg gcttctgg 3814333DNAArtificial sequenceExemplary element X variant 143gtacttcata cttttcattc caatggcttc tgg 3314433DNAArtificial sequenceExemplary element X variant 144gtactctttt cattccaatg actttgcttc tgg 3314533DNAArtificial sequenceExemplary element X variant 145gtacttcata cattccaatg actttgcttc tgg 3314628DNAArtificial sequenceExemplary element X variant 146gtactcattc caatgacttt gcttctgg 2814733DNAArtificial sequenceExemplary element X variant 147gtactctttt cattccaatg gggtggcttc tgg 3314833DNAArtificial sequenceExemplary element X variant 148gtacttcata cattccaatg gggtggcttc tgg 3314928DNAArtificial sequenceExemplary element X variant 149gtactcattc caatggggtg gcttctgg 2815028DNAArtificial sequenceExemplary element X variant 150gtactctttt cattccaatg gcttctgg 2815128DNAArtificial sequenceExemplary element X variant 151gtacttcata cattccaatg gcttctgg 2815223DNAArtificial sequenceExemplary element X variant 152gtactcattc caatggcttc tgg 2315343DNAArtificial sequenceExemplary element X variant 153gtacttcata cttttcattc caatggggtg actttgcttc tgg 4315419DNAArtificial sequenceExemplary element X variant 154acttttcatt ccaatgggg 1915519DNAArtificial sequenceExemplary element X variant 155cttttcattc caatggggt 1915619DNAArtificial sequenceExemplary element X variant 156ttttcattcc aatggggtg 1915719DNAArtificial sequenceExemplary element X variant 157tttcattcca atggggtga 1915819DNAArtificial sequenceExemplary element X variant 158tacttttcat tccaatggg 1915919DNAArtificial sequenceExemplary element X variant 159atacttttca ttccaatgg 1916019DNAArtificial sequenceExemplary element X variant 160catacttttc attccaatg 1916119DNAArtificial sequenceExemplary element X variant 161tcatactttt cattccaat 1916219DNAArtificial sequenceExemplary element X variant 162ttcatacttt tcattccaa 1916319DNAArtificial sequenceExemplary element X variant 163ttccaatggg gtgactttg 1916419DNAArtificial sequenceExemplary element X variant 164attccaatgg ggtgacttt 1916519DNAArtificial sequenceExemplary element X variant 165cattccaatg gggtgactt 1916619DNAArtificial sequenceExemplary element X variant 166tcattccaat ggggtgact 1916719DNAArtificial sequenceExemplary element X variant 167ttcattccaa tggggtgac 1916819DNAArtificial sequenceExemplary element X variant 168tttcattcca atggggtga 1916919DNAArtificial sequenceExemplary element X variant 169ttttcattcc aatggggtg 1917019DNAArtificial sequenceExemplary element X variant 170cttttcattc caatggggt 1917119DNAArtificial sequenceExemplary element X variant 171acttttcatt ccaatgggg 1917224DNAArtificial sequenceExemplary element X variant 172tttcattcca atggggtgac tttg 2417324DNAArtificial sequenceExemplary element X variant 173ttttcattcc aatggggtga cttt 2417424DNAArtificial sequenceExemplary element X variant 174cttttcattc caatggggtg actt 2417524DNAArtificial sequenceExemplary element X variant 175acttttcatt ccaatggggt gact 2417624DNAArtificial sequenceExemplary element X variant 176tacttttcat tccaatgggg tgac 2417724DNAArtificial sequenceExemplary element X variant 177atacttttca ttccaatggg gtga 2417824DNAArtificial sequenceExemplary element X variant 178catacttttc attccaatgg ggtg 2417924DNAArtificial sequenceExemplary element X variant 179tcatactttt cattccaatg gggt 2418024DNAArtificial sequenceExemplary element X variant 180ttcatacttt tcattccaat gggg 2418119DNAArtificial sequenceExemplary element X variant 181ttttcattcc aatggggtg 1918219DNAArtificial sequenceExemplary element X variant 182tttcattcca atggggtga 1918319DNAArtificial sequenceExemplary element X variant 183attccaatgg ggtgacttt 1918419DNAArtificial sequenceExemplary element X variant 184ttccaatggg gtgactttg 1918519DNAArtificial sequenceExemplary element X variant 185cattccaatg gggtgactt 1918619DNAArtificial sequenceExemplary element X variant 186tcattccaat ggggtgact 1918719DNAArtificial sequenceExemplary element X variant 187ttcattccaa

tggggtgac 1918819DNAArtificial sequenceExemplary element X variant 188tttcattcca atggggtga 1918919DNAArtificial sequenceExemplary element X variant 189ttttcattcc aatggggtg 1919024DNAArtificial sequenceExemplary element X variant 190ttttcattcc aatggggtga cttt 2419125DNAArtificial sequenceExemplary element X variant 191tcatactttt cattccaatg gggtg 2519223DNAArtificial sequenceExemplary element X variant 192tttcattcca atggggtgac ttt 2319319DNAArtificial sequenceExemplary element X variant 193ttttcattcg ggtgacttt 1919424DNAArtificial sequenceExemplary element X variant 194ttttcattcc aatgactttg cttc 2419519DNAArtificial sequenceExemplary element X variant 195ttttcattca ctttgcttc 1919619DNAArtificial sequenceExemplary element X variant 196attccaatgg ggtgacttt 1919719DNAArtificial sequenceExemplary element X variant 197attccaatga ctttgcttc 1919819DNAArtificial sequenceExemplary element X variant 198ttccaatgac tttgcttct 1919924DNAArtificial sequenceExemplary element X variant 199tttcattcca atggggtgac tttg 2420024DNAArtificial sequenceExemplary element X variant 200ttttcattcc aatggggtga cttt 2420125DNAArtificial sequenceExemplary element X variant 201cttttcattc caatggggtg acttt 2520224DNAArtificial sequenceExemplary element X variant 202ttttcattcc aatgactttg cttc 2420323DNAArtificial sequenceExemplary element X variant 203tttcattcca atggggtgac ttt 2320419DNAArtificial sequenceExemplary element X variant 204ttttcattcg ggtgacttt 1920519DNAArtificial sequenceExemplary element X variant 205tttcattcgg gtgactttg 1920619DNAArtificial sequenceExemplary element X variant 206attccaatgg ggtgacttt 1920719DNAArtificial sequenceExemplary element X variant 207ttccaatggg gtgactttg 1920824DNAArtificial sequenceExemplary element X variant 208ttttcattcc aatggggtga cttt 2420924DNAArtificial sequenceExemplary element X variant 209tttcattcca atggggtgac tttg 2421024DNAArtificial sequenceExemplary element X variant 210tttcattcca atggggtgac tttg 2421124DNAArtificial sequenceExemplary element X variant 211ttttcattcc aatggggtga cttt 2421224DNAArtificial sequenceExemplary element X variant 212cttttcattc caatggggtg actt 2421324DNAArtificial sequenceExemplary element X variant 213acttttcatt ccaatggggt gact 2421424DNAArtificial sequenceExemplary element X variant 214ttccaatggg gtgactttgc ttct 2421524DNAArtificial sequenceExemplary element X variant 215attccaatgg ggtgactttg cttc 2421624DNAArtificial sequenceExemplary element X variant 216cattccaatg gggtgacttt gctt 2421724DNAArtificial sequenceExemplary element X variant 217tcattccaat ggggtgactt tgct 2421824DNAArtificial sequenceExemplary element X variant 218ttcattccaa tggggtgact ttgc 2421924DNAArtificial sequenceExemplary element X variant 219tttcattcca atggggtgac tttg 2422024DNAArtificial sequenceExemplary element X variant 220ttttcattcc aatggggtga cttt 2422124DNAArtificial sequenceExemplary element X variant 221cttttcattc caatggggtg actt 2422224DNAArtificial sequenceExemplary element X variant 222acttttcatt ccaatggggt gact 2422329DNAArtificial sequenceExemplary element X variant 223tttcattcca atggggtgac tttgcttct 2922429DNAArtificial sequenceExemplary element X variant 224ttttcattcc aatggggtga ctttgcttc 2922529DNAArtificial sequenceExemplary element X variant 225cttttcattc caatggggtg actttgctt 2922629DNAArtificial sequenceExemplary element X variant 226acttttcatt ccaatggggt gactttgct 2922729DNAArtificial sequenceExemplary element X variant 227tacttttcat tccaatgggg tgactttgc 2922829DNAArtificial sequenceExemplary element X variant 228atacttttca ttccaatggg gtgactttg 2922929DNAArtificial sequenceExemplary element X variant 229catacttttc attccaatgg ggtgacttt 2923029DNAArtificial sequenceExemplary element X variant 230tcatactttt cattccaatg gggtgactt 2923129DNAArtificial sequenceExemplary element X variant 231ttcatacttt tcattccaat ggggtgact 2923226DNAArtificial sequenceExemplary element X variant 232tttcattcca atggggtgac tttgct 2623324DNAArtificial sequenceExemplary element X variant 233cttttcattc caatgacttt gctt 2423424DNAArtificial sequenceExemplary element X variant 234cttttcattc gggtgacttt gctt 2423524DNAArtificial sequenceExemplary element X variant 235ttttcattcc aatgactttg cttc 2423624DNAArtificial sequenceExemplary element X variant 236ttttcattcg ggtgactttg cttc 2423724DNAArtificial sequenceExemplary element X variant 237attccaatgg ggtgactttg cttc 2423824DNAArtificial sequenceExemplary element X variant 238ttccaatggg gtgactttgc ttct 2423924DNAArtificial sequenceExemplary element X variant 239cattccaatg gggtgacttt gctt 2424024DNAArtificial sequenceExemplary element X variant 240tcattccaat ggggtgactt tgct 2424129DNAArtificial sequenceExemplary element X variant 241tttcattcca atggggtgac tttgcttct 2924229DNAArtificial sequenceExemplary element X variant 242ttttcattcc aatggggtga ctttgcttc 2924329DNAArtificial sequenceExemplary element X variant 243cttttcattc caatggggtg actttgctt 2924429DNAArtificial sequenceExemplary element X variant 244tcatactttt cattccaatg actttgctt 2924529DNAArtificial sequenceExemplary element X variant 245catacttttc attccaatga ctttgcttc 2924629DNAArtificial sequenceExemplary element X variant 246atacttttca ttccaatgac tttgcttct 2924729DNAArtificial sequenceExemplary element X variant 247tacttttcat tccaatgact ttgcttctg 2924829DNAArtificial sequenceExemplary element X variant 248acttttcatt ccaatgactt tgcttctgg 2924929DNAArtificial sequenceExemplary element X variant 249tttcattcca atggggtgac tttgcttct 2925029DNAArtificial sequenceExemplary element X variant 250ttttcattcc aatggggtga ctttgcttc 2925129DNAArtificial sequenceExemplary element X variant 251cttttcattc caatggggtg actttgctt 2925229DNAArtificial sequenceExemplary element X variant 252acttttcatt ccaatggggt gactttgct 2925329DNAArtificial sequenceExemplary element X variant 253ttcatacttt tcattcgggt gactttgct 2925429DNAArtificial sequenceExemplary element X variant 254tcatactttt cattcgggtg actttgctt 2925529DNAArtificial sequenceExemplary element X variant 255catacttttc attcgggtga ctttgcttc 2925629DNAArtificial sequenceExemplary element X variant 256atacttttca ttcgggtgac tttgcttct 2925729DNAArtificial sequenceExemplary element X variant 257tacttttcat tcgggtgact ttgcttctg 2925829DNAArtificial sequenceExemplary element X variant 258acttttcatt cgggtgactt tgcttctgg 2925929DNAArtificial sequenceExemplary element X variant 259acttttcatt ccaatggggt gactttgct 2926029DNAArtificial sequenceExemplary element X variant 260cttttcattc caatggggtg actttgctt 2926129DNAArtificial sequenceExemplary element X variant 261ttttcattcc aatggggtga ctttgcttc 2926229DNAArtificial sequenceExemplary element X variant 262tttcattcca atggggtgac tttgcttct 2926329DNAArtificial sequenceExemplary element X variant 263ttcattccaa tggggtgact ttgcttctg 2926429DNAArtificial sequenceExemplary element X variant 264tcattccaat ggggtgactt tgcttctgg 2926534DNAArtificial sequenceExemplary element X variant 265ttcatacttt tcattccaat ggggtgactt tgct 3426634DNAArtificial sequenceExemplary element X variant 266tcatactttt cattccaatg gggtgacttt gctt 3426734DNAArtificial sequenceExemplary element X variant 267catacttttc attccaatgg ggtgactttg cttc 3426834DNAArtificial sequenceExemplary element X variant 268atacttttca ttccaatggg gtgactttgc ttct 3426934DNAArtificial sequenceExemplary element X variant 269tacttttcat tccaatgggg tgactttgct tctg 3427034DNAArtificial sequenceExemplary element X variant 270acttttcatt ccaatggggt gactttgctt ctgg 3427128DNAArtificial sequenceExemplary element X variant 271tttcattcca atggggtgac tttgcttc 2827224DNAArtificial sequenceExemplary element X variant 272ttttcattcg ggtgactttg cttc 2427324DNAArtificial sequenceExemplary element X variant 273ttttcattcc aatgactttg cttc 2427424DNAArtificial sequenceExemplary element X variant 274ttttcattcc aatggggtgg cttc 2427525DNAArtificial sequenceExemplary element X variant 275cttttcattc caatggggtg gcttc 2527624DNAArtificial sequenceExemplary element X variant 276catacttttc attcgggtgg cttc 2427724DNAArtificial sequenceExemplary element X variant 277catacttttc attcactttg cttc 2427824DNAArtificial sequenceExemplary element X variant 278catacttttc attccaatgg cttc 2427924DNAArtificial sequenceExemplary element X variant 279tacttcatac ttttcattcg cttc 2428023DNAArtificial sequenceExemplary element X variant 280ttccaatggg gtgactttgc ttc 2328119DNAArtificial sequenceExemplary element X variant 281ttccaatgac tttgcttct 1928219DNAArtificial sequenceExemplary element X variant 282ttccaatggg gtggcttct 1928319DNAArtificial sequenceExemplary element X variant 283tttcattcca atggcttct 1928419DNAArtificial sequenceExemplary element X variant 284ttttcattcc aatggcttc 1928524DNAArtificial sequenceExemplary element X variant 285attccaatgg ggtgactttg cttc 2428620DNAArtificial sequenceExemplary element X variant 286attccaatga ctttgcttct 2028720DNAArtificial sequenceExemplary element X variant 287attccaatgg ggtggcttct 2028828DNAArtificial sequenceExemplary element X variant 288tttcattcca atggggtgac tttgcttc 2828924DNAArtificial sequenceExemplary element X variant 289ttttcattcc aatgactttg cttc 2429024DNAArtificial sequenceExemplary element X variant 290ttttcattcc aatggggtgg cttc 2429124DNAArtificial sequenceExemplary element X variant 291catacttttc attccaatgg cttc 2429228DNAArtificial sequenceExemplary element X variant 292tttcattcca atggggtgac tttgcttc 2829324DNAArtificial sequenceExemplary element X variant 293ttttcattcc aatgactttg cttc 2429428DNAArtificial sequenceExemplary element X variant 294tttcattcca atggggtgac tttgcttc 2829523DNAArtificial sequenceExemplary element X variant 295tttcattcca atggggtggc ttc 2329623DNAArtificial sequenceExemplary element X variant 296tttcattcgg gtgactttgc ttc 2329719DNAArtificial sequenceExemplary element X variant 297tttcattcgg gtggcttct 1929819DNAArtificial sequenceExemplary element X variant 298ttttcattcg ggtggcttc 1929923DNAArtificial sequenceExemplary element X variant 299ttccaatggg gtgactttgc ttc 2330019DNAArtificial sequenceExemplary element X variant 300attccaatgg ggtggcttc 1930119DNAArtificial sequenceExemplary element X variant 301attcaatggg gtggcttct 1930228DNAArtificial sequenceExemplary element X variant 302tttcattcca atggggtgac tttgcttc 2830324DNAArtificial sequenceExemplary element X variant 303ttttcattcc aatggggtgg cttc 2430428DNAArtificial sequenceExemplary element X variant 304tttcattcca atggggtgac tttgcttc 2830523DNAArtificial sequenceExemplary element X variant 305tttcattcgg gtgactttgc ttc 2330623DNAArtificial sequenceExemplary element X variant 306tttcattcca atgactttgc ttc 2330719DNAArtificial sequenceExemplary element X variant 307ttttcattca ctttgcttc 1930819DNAArtificial sequenceExemplary element X variant 308tttcattcac tttgcttct 1930923DNAArtificial sequenceExemplary element X variant 309ttccaatggg gtgactttgc ttc 2331019DNAArtificial sequenceExemplary element X variant 310ttccaatgac tttgcttct 1931119DNAArtificial sequenceExemplary element X variant 311attccaatga ctttgcttc 1931228DNAArtificial sequenceExemplary element X variant 312tttcattcca atggggtgac tttgcttc

2831324DNAArtificial sequenceExemplary element X variant 313ttttcattcc aatgactttg cttc 2431424DNAArtificial sequenceExemplary element X variant 314tttcattcca atgactttgc ttct 2431528DNAArtificial sequenceExemplary element X variant 315tttcattcca atggggtgac tttgcttc 2831623DNAArtificial sequenceExemplary element X variant 316tttcattcgg gtgactttgc ttc 2331723DNAArtificial sequenceExemplary element X variant 317ttccaatggg gtgactttgc ttc 2331828DNAArtificial sequenceExemplary element X variant 318tttcattcca atggggtgac tttgcttc 2831928DNAArtificial sequenceExemplary element X variant 319tttcattcca atggggtgac tttgcttc 2832024DNAArtificial sequenceExemplary element X variant 320ttttcattcg ggtgactttg cttc 2432124DNAArtificial sequenceExemplary element X variant 321ttttcattcc aatggggtgg cttc 2432224DNAArtificial sequenceExemplary element X variant 322catacttttc attcgggtgg cttc 2432324DNAArtificial sequenceExemplary element X variant 323tttcattcgg gtgactttgc ttct 2432424DNAArtificial sequenceExemplary element X variant 324tttcattcca atggggtggc ttct 2432524DNAArtificial sequenceExemplary element X variant 325atacttttca ttcgggtggc ttct 2432624DNAArtificial sequenceExemplary element X variant 326tacttttcat tcgggtggct tctg 2432724DNAArtificial sequenceExemplary element X variant 327acttttcatt cgggtggctt ctgg 2432824DNAArtificial sequenceExemplary element X variant 328ttccaatggg gtgactttgc ttct 2432924DNAArtificial sequenceExemplary element X variant 329attccaatgg ggtgactttg cttc 2433024DNAArtificial sequenceExemplary element X variant 330ttttcattcc aatggggtgg cttc 2433124DNAArtificial sequenceExemplary element X variant 331tttcattcca atggggtggc ttct 2433224DNAArtificial sequenceExemplary element X variant 332ttcattccaa tggggtggct tctg 2433329DNAArtificial sequenceExemplary element X variant 333tcattccaat ggggtgactt tgcttctgg 2933428DNAArtificial sequenceExemplary element X variant 334tttcattcca atggggtgac tttgcttc 2833530DNAArtificial sequenceExemplary element X variant 335ttttcattcc aatggggtga ctttgcttct 3033629DNAArtificial sequenceExemplary element X variant 336atacttttca ttccaatggg gtggcttct 2933729DNAArtificial sequenceExemplary element X variant 337catacttttc attccaatgg ggtggcttc 2933828DNAArtificial sequenceExemplary element X variant 338tttcattcca atggggtgac tttgcttc 2833924DNAArtificial sequenceExemplary element X variant 339ttttcattcc aatgactttg cttc 2434024DNAArtificial sequenceExemplary element X variant 340ttttcattcg ggtgactttg cttc 2434124DNAArtificial sequenceExemplary element X variant 341catacttttc attcactttg cttc 2434224DNAArtificial sequenceExemplary element X variant 342atacttttca ttcactttgc ttct 2434324DNAArtificial sequenceExemplary element X variant 343tacttttcat tcactttgct tctg 2434424DNAArtificial sequenceExemplary element X variant 344acttttcatt cactttgctt ctgg 2434524DNAArtificial sequenceExemplary element X variant 345ttccaatggg gtgactttgc ttct 2434624DNAArtificial sequenceExemplary element X variant 346attccaatgg ggtgactttg cttc 2434724DNAArtificial sequenceExemplary element X variant 347tttcattcca atgactttgc ttct 2434824DNAArtificial sequenceExemplary element X variant 348ttttcattcc aatgactttg cttc 2434929DNAArtificial sequenceExemplary element X variant 349ttttcattcc aatggggtga ctttgcttc 2935029DNAArtificial sequenceExemplary element X variant 350tttcattcca atggggtgac tttgcttct 2935129DNAArtificial sequenceExemplary element X variant 351catacttttc attccaatga ctttgcttc 2935229DNAArtificial sequenceExemplary element X variant 352atacttttca ttccaatgac tttgcttct 2935329DNAArtificial sequenceExemplary element X variant 353tacttttcat tccaatgact ttgcttctg 2935429DNAArtificial sequenceExemplary element X variant 354acttttcatt ccaatgactt tgcttctgg 2935529DNAArtificial sequenceExemplary element X variant 355ttttcattcc aatggggtga ctttgcttc 2935629DNAArtificial sequenceExemplary element X variant 356ttttcattcc aatggggtga ctttgcttc 2935724DNAArtificial sequenceExemplary element X variant 357tttcattcgg gtgactttgc ttct 2435829DNAArtificial sequenceExemplary element X variant 358catacttttc attcgggtga ctttgcttc 2935929DNAArtificial sequenceExemplary element X variant 359atacttttca ttcgggtgac tttgcttct 2936029DNAArtificial sequenceExemplary element X variant 360tacttttcat tcgggtgact ttgcttctg 2936129DNAArtificial sequenceExemplary element X variant 361acttttcatt cgggtgactt tgcttctgg 2936229DNAArtificial sequenceExemplary element X variant 362ttttcattcc aatggggtga ctttgcttc 2936329DNAArtificial sequenceExemplary element X variant 363tttcattcca atggggtgac tttgcttct 2936429DNAArtificial sequenceExemplary element X variant 364ttcattccaa tggggtgact ttgcttctg 2936529DNAArtificial sequenceExemplary element X variant 365tcattccaat ggggtgactt tgcttctgg 2936634DNAArtificial sequenceExemplary element X variant 366acttttcatt ccaatggggt gactttgctt ctgg 3436734DNAArtificial sequenceExemplary element X variant 367tacttttcat tccaatgggg tgactttgct tctg 3436834DNAArtificial sequenceExemplary element X variant 368atacttttca ttccaatggg gtgactttgc ttct 3436934DNAArtificial sequenceExemplary element X variant 369catacttttc attccaatgg ggtgactttg cttc 3437025DNAArtificial sequenceExemplary element X variant 370cttttcattc caatggggtg acttt 2537125DNAArtificial sequenceExemplary element X variant 371acttttcatt ccaatggggt gactt 2537225DNAArtificial sequenceExemplary element X variant 372tacttttcat tccaatgggg tgact 2537325DNAArtificial sequenceExemplary element X variant 373atacttttca ttccaatggg gtgac 2537425DNAArtificial sequenceExemplary element X variant 374catacttttc attccaatgg ggtga 2537525DNAArtificial sequenceExemplary element X variant 375tcatactttt cattccaatg gggtg 2537625DNAArtificial sequenceExemplary element X variant 376ttcatacttt tcattccaat ggggt 2537725DNAArtificial sequenceExemplary element X variant 377cttcatactt ttcattccaa tgggg 2537819DNAArtificial sequenceExemplary element X variant 378ttcatacttt tcattccaa 1937919DNAArtificial sequenceExemplary element X variant 379tcatactttt cattccaat 1938019DNAArtificial sequenceExemplary element X variant 380catacttttc attccaatg 1938119DNAArtificial sequenceExemplary element X variant 381atacttttca ttccaatgg 1938219DNAArtificial sequenceExemplary element X variant 382tacttttcat tccaatggg 1938319DNAArtificial sequenceExemplary element X variant 383acttttcatt ccaatgggg 1938419DNAArtificial sequenceExemplary element X variant 384cttttcattc caatggggt 1938520DNAArtificial sequenceExemplary element X variant 385cttttcattc caatggggtg 2038620DNAArtificial sequenceExemplary element X variant 386catacttttc attcgggtga 2038720DNAArtificial sequenceExemplary element X variant 387atacttttca ttcgggtgac 2038820DNAArtificial sequenceExemplary element X variant 388tacttttcat tcgggtgact 2038920DNAArtificial sequenceExemplary element X variant 389acttttcatt cgggtgactt 2039020DNAArtificial sequenceExemplary element X variant 390cttttcattc gggtgacttt 2039120DNAArtificial sequenceExemplary element X variant 391cttttcattc caatggggtg 2039219DNAArtificial sequenceExemplary element X variant 392catacttttc aatggggtg 1939319DNAArtificial sequenceExemplary element X variant 393atacttttca atggggtga 1939419DNAArtificial sequenceExemplary element X variant 394tacttttcaa tggggtgac 1939519DNAArtificial sequenceExemplary element X variant 395acttttcaat ggggtgact 1939619DNAArtificial sequenceExemplary element X variant 396cttttcaatg gggtgactt 1939724DNAArtificial sequenceExemplary element X variant 397catacttttc attccaatgg ggtg 2439824DNAArtificial sequenceExemplary element X variant 398atacttttca ttccaatggg gtga 2439924DNAArtificial sequenceExemplary element X variant 399tacttttcat tccaatgggg tgac 2440024DNAArtificial sequenceExemplary element X variant 400acttttcatt ccaatggggt gact 2440124DNAArtificial sequenceExemplary element X variant 401cttttcattc caatggggtg actt 2440225DNAArtificial sequenceExemplary element X variant 402cttttcattc caatggggtg acttt 2540320DNAArtificial sequenceExemplary element X variant 403cttttcattc caatgacttt 2040420DNAArtificial sequenceExemplary element X variant 404cttttcattc gggtgacttt 2040519DNAArtificial sequenceExemplary element X variant 405catacttttc attcacttt 1940619DNAArtificial sequenceExemplary element X variant 406atacttttca ttcactttg 1940719DNAArtificial sequenceExemplary element X variant 407tacttttcat tcactttgc 1940819DNAArtificial sequenceExemplary element X variant 408acttttcatt cactttgct 1940919DNAArtificial sequenceExemplary element X variant 409cttttcattc actttgctt 1941025DNAArtificial sequenceExemplary element X variant 410cttttcattc caatggggtg acttt 2541120DNAArtificial sequenceExemplary element X variant 411cttttcattc caatgacttt 2041220DNAArtificial sequenceExemplary element X variant 412cttttcaatg gggtgacttt 2041319DNAArtificial sequenceExemplary element X variant 413catacttttc aatgacttt 1941419DNAArtificial sequenceExemplary element X variant 414atacttttca atgactttg 1941519DNAArtificial sequenceExemplary element X variant 415tacttttcaa tgactttgc 1941619DNAArtificial sequenceExemplary element X variant 416acttttcaat gactttgct 1941719DNAArtificial sequenceExemplary element X variant 417cttttcaatg actttgctt 1941825DNAArtificial sequenceExemplary element X variant 418cttttcattc caatggggtg acttt 2541924DNAArtificial sequenceExemplary element X variant 419catacttttc attccaatga cttt 2442024DNAArtificial sequenceExemplary element X variant 420atacttttca ttccaatgac tttg 2442124DNAArtificial sequenceExemplary element X variant 421tacttttcat tccaatgact ttgc 2442224DNAArtificial sequenceExemplary element X variant 422acttttcatt ccaatgactt tgct 2442324DNAArtificial sequenceExemplary element X variant 423cttttcattc caatgacttt gctt 2442425DNAArtificial sequenceExemplary element X variant 424cttttcattc caatggggtg acttt 2542520DNAArtificial sequenceExemplary element X variant 425cttttcattc gggtgacttt 2042625DNAArtificial sequenceExemplary element X variant 426cttttcattc caatggggtg acttt 2542724DNAArtificial sequenceExemplary element X variant 427cttttcattc caatggggtg actt 2442824DNAArtificial sequenceExemplary element X variant 428acttttcatt ccaatggggt gact 2442924DNAArtificial sequenceExemplary element X variant 429ttcatacttt tcattcgggt gact 2443024DNAArtificial sequenceExemplary element X variant 430tcatactttt cattcgggtg actt 2443124DNAArtificial sequenceExemplary element X variant 431catacttttc attcgggtga cttt 2443224DNAArtificial sequenceExemplary element X variant 432atacttttca ttcgggtgac tttg 2443324DNAArtificial sequenceExemplary element X variant 433tacttttcat tcgggtgact ttgc 2443424DNAArtificial sequenceExemplary element X variant 434acttttcatt cgggtgactt tgct 2443524DNAArtificial sequenceExemplary element X variant 435cttttcattc gggtgacttt gctt 2443624DNAArtificial sequenceExemplary element X variant 436cttttcattc caatggggtg actt 2443724DNAArtificial sequenceExemplary element X variant 437acttttcatt ccaatggggt gact 2443824DNAArtificial sequenceExemplary element X variant 438ttcatacttt

tcaatggggt gact 2443924DNAArtificial sequenceExemplary element X variant 439tcatactttt caatggggtg actt 2444024DNAArtificial sequenceExemplary element X variant 440catacttttc aatggggtga cttt 2444124DNAArtificial sequenceExemplary element X variant 441atacttttca atggggtgac tttg 2444224DNAArtificial sequenceExemplary element X variant 442tacttttcaa tggggtgact ttgc 2444324DNAArtificial sequenceExemplary element X variant 443acttttcaat ggggtgactt tgct 2444424DNAArtificial sequenceExemplary element X variant 444cttttcaatg gggtgacttt gctt 2444529DNAArtificial sequenceExemplary element X variant 445ttcatacttt tcattccaat ggggtgact 2944629DNAArtificial sequenceExemplary element X variant 446tcatactttt cattccaatg gggtgactt 2944729DNAArtificial sequenceExemplary element X variant 447catacttttc attccaatgg ggtgacttt 2944829DNAArtificial sequenceExemplary element X variant 448atacttttca ttccaatggg gtgactttg 2944929DNAArtificial sequenceExemplary element X variant 449tacttttcat tccaatgggg tgactttgc 2945029DNAArtificial sequenceExemplary element X variant 450acttttcatt ccaatggggt gactttgct 2945129DNAArtificial sequenceExemplary element X variant 451cttttcattc caatggggtg actttgctt 2945228DNAArtificial sequenceExemplary element X variant 452cttttcattc caatggggtg actttgct 2845324DNAArtificial sequenceExemplary element X variant 453cttttcattc gggtgacttt gctt 2445424DNAArtificial sequenceExemplary element X variant 454cttttcattc caatgacttt gctt 2445524DNAArtificial sequenceExemplary element X variant 455tcatactttt cattcacttt gctt 2445624DNAArtificial sequenceExemplary element X variant 456acttttcatt cgggtgactt tgct 2445724DNAArtificial sequenceExemplary element X variant 457acttttcatt ccaatgactt tgct 2445824DNAArtificial sequenceExemplary element X variant 458ttcatacttt tcattcactt tgct 2445928DNAArtificial sequenceExemplary element X variant 459cttttcattc caatggggtg actttgct 2846024DNAArtificial sequenceExemplary element X variant 460cttttcaatg gggtgacttt gctt 2446124DNAArtificial sequenceExemplary element X variant 461cttttcattc caatgacttt gctt 2446224DNAArtificial sequenceExemplary element X variant 462tcatactttt caatgacttt gctt 2446324DNAArtificial sequenceExemplary element X variant 463acttttcaat ggggtgactt tgct 2446424DNAArtificial sequenceExemplary element X variant 464acttttcatt ccaatgactt tgct 2446524DNAArtificial sequenceExemplary element X variant 465ttcatacttt tcaatgactt tgct 2446629DNAArtificial sequenceExemplary element X variant 466cttttcattc caatggggtg actttgctt 2946729DNAArtificial sequenceExemplary element X variant 467acttttcatt ccaatggggt gactttgct 2946829DNAArtificial sequenceExemplary element X variant 468acttttcatt ccaatgactt tgcttctgg 2946929DNAArtificial sequenceExemplary element X variant 469cttttcattc caatgacttt gcttctgga 2947029DNAArtificial sequenceExemplary element X variant 470tcatactttt cattccaatg actttgctt 2947129DNAArtificial sequenceExemplary element X variant 471ttcatacttt tcattccaat gactttgct 2947229DNAArtificial sequenceExemplary element X variant 472cttttcattc caatggggtg actttgctt 2947329DNAArtificial sequenceExemplary element X variant 473acttttcatt ccaatggggt gactttgct 2947429DNAArtificial sequenceExemplary element X variant 474acttttcaat ggggtgactt tgcttctgg 2947529DNAArtificial sequenceExemplary element X variant 475cttttcaatg gggtgacttt gcttctgga 2947629DNAArtificial sequenceExemplary element X variant 476tcatactttt caatggggtg actttgctt 2947729DNAArtificial sequenceExemplary element X variant 477ttcatacttt tcaatggggt gactttgct 2947834DNAArtificial sequenceExemplary element X variant 478acttttcatt ccaatggggt gactttgctt ctgg 3447934DNAArtificial sequenceExemplary element X variant 479tacttttcat tccaatgggg tgactttgct tctg 3448034DNAArtificial sequenceExemplary element X variant 480atacttttca ttccaatggg gtgactttgc ttct 3448134DNAArtificial sequenceExemplary element X variant 481catacttttc attccaatgg ggtgactttg cttc 3448234DNAArtificial sequenceExemplary element X variant 482tcatactttt cattccaatg gggtgacttt gctt 3448334DNAArtificial sequenceExemplary element X variant 483ttcatacttt tcattccaat ggggtgactt tgct 3448430DNAArtificial sequenceExemplary element X variant 484cttttcattc caatggggtg actttgcttc 3048525DNAArtificial sequenceExemplary element X variant 485cttttcattc gggtgacttt gcttc 2548625DNAArtificial sequenceExemplary element X variant 486cttttcattc caatgacttt gcttc 2548725DNAArtificial sequenceExemplary element X variant 487cttttcattc caatggggtg gcttc 2548820DNAArtificial sequenceExemplary element X variant 488cttttcattc actttgcttc 2048920DNAArtificial sequenceExemplary element X variant 489cttttcattc caatggcttc 2049020DNAArtificial sequenceExemplary element X variant 490cttttcattc gggtggcttc 2049119DNAArtificial sequenceExemplary element X variant 491cttttcattc gcttctgga 1949219DNAArtificial sequenceExemplary element X variant 492acttttcatt cgcttctgg 1949319DNAArtificial sequenceExemplary element X variant 493tacttttcat tcgcttctg 1949419DNAArtificial sequenceExemplary element X variant 494atacttttca ttcgcttct 1949519DNAArtificial sequenceExemplary element X variant 495catacttttc attcgcttc 1949625DNAArtificial sequenceExemplary element X variant 496cttttcaatg gggtgacttt gcttc 2549725DNAArtificial sequenceExemplary element X variant 497cttttcattc caatgacttt gcttc 2549825DNAArtificial sequenceExemplary element X variant 498cttttcattc caatggggtg gcttc 2549920DNAArtificial sequenceExemplary element X variant 499cttttcaatg actttgcttc 2050020DNAArtificial sequenceExemplary element X variant 500cttttcaatg gggtggcttc 2050120DNAArtificial sequenceExemplary element X variant 501cttttcattc caatggcttc 2050230DNAArtificial sequenceExemplary element X variant 502cttttcattc caatggggtg actttgcttc 3050319DNAArtificial sequenceExemplary element X variant 503cttttcaatg gcttctgga 1950419DNAArtificial sequenceExemplary element X variant 504acttttcaat ggcttctgg 1950519DNAArtificial sequenceExemplary element X variant 505tacttttcaa tggcttctg 1950619DNAArtificial sequenceExemplary element X variant 506atacttttca atggcttct 1950719DNAArtificial sequenceExemplary element X variant 507catacttttc aatggcttc 1950830DNAArtificial sequenceExemplary element X variant 508cttttcattc caatggggtg actttgcttc 3050925DNAArtificial sequenceExemplary element X variant 509cttttcattc caatgacttt gcttc 2551025DNAArtificial sequenceExemplary element X variant 510cttttcattc caatggggtg gcttc 2551124DNAArtificial sequenceExemplary element X variant 511cttttcattc caatggcttc tgga 2451224DNAArtificial sequenceExemplary element X variant 512acttttcatt ccaatggctt ctgg 2451324DNAArtificial sequenceExemplary element X variant 513tacttttcat tccaatggct tctg 2451424DNAArtificial sequenceExemplary element X variant 514atacttttca ttccaatggc ttct 2451524DNAArtificial sequenceExemplary element X variant 515catacttttc attccaatgg cttc 2451630DNAArtificial sequenceExemplary element X variant 516cttttcattc caatggggtg actttgcttc 3051725DNAArtificial sequenceExemplary element X variant 517cttttcattc gggtgacttt gcttc 2551825DNAArtificial sequenceExemplary element X variant 518cttttcattc caatggggtg gcttc 2551920DNAArtificial sequenceExemplary element X variant 519cttttcattc gggtggcttc 2052030DNAArtificial sequenceExemplary element X variant 520cttttcattc caatggggtg actttgcttc 3052125DNAArtificial sequenceExemplary element X variant 521cttttcaatg gggtgacttt gcttc 2552225DNAArtificial sequenceExemplary element X variant 522cttttcattc caatggggtg gcttc 2552320DNAArtificial sequenceExemplary element X variant 523cttttcaatg gggtggcttc 2052430DNAArtificial sequenceExemplary element X variant 524cttttcattc caatggggtg actttgcttc 3052525DNAArtificial sequenceExemplary element X variant 525cttttcattc caatggggtg gcttc 2552630DNAArtificial sequenceExemplary element X variant 526cttttcattc caatggggtg actttgcttc 3052725DNAArtificial sequenceExemplary element X variant 527cttttcattc gggtgacttt gcttc 2552825DNAArtificial sequenceExemplary element X variant 528cttttcattc caatgacttt gcttc 2552920DNAArtificial sequenceExemplary element X variant 529cttttcattc actttgcttc 2053030DNAArtificial sequenceExemplary element X variant 530cttttcattc caatggggtg actttgcttc 3053125DNAArtificial sequenceExemplary element X variant 531cttttcaatg gggtgacttt gcttc 2553225DNAArtificial sequenceExemplary element X variant 532cttttcattc caatgacttt gcttc 2553320DNAArtificial sequenceExemplary element X variant 533cttttcaatg actttgcttc 2053430DNAArtificial sequenceExemplary element X variant 534cttttcattc caatggggtg actttgcttc 3053525DNAArtificial sequenceExemplary element X variant 535cttttcattc caatgacttt gcttc 2553630DNAArtificial sequenceExemplary element X variant 536cttttcattc caatggggtg actttgcttc 3053725DNAArtificial sequenceExemplary element X variant 537cttttcattc gggtgacttt gcttc 2553830DNAArtificial sequenceExemplary element X variant 538cttttcattc caatggggtg actttgcttc 3053925DNAArtificial sequenceExemplary element X variant 539cttttcaatg gggtgacttt gcttc 2554030DNAArtificial sequenceExemplary element X variant 540cttttcattc caatggggtg actttgcttc 3054130DNAArtificial sequenceExemplary element X variant 541cttttcattc caatggggtg actttgcttc 3054225DNAArtificial sequenceExemplary element X variant 542cttttcattc gggtgacttt gcttc 2554325DNAArtificial sequenceExemplary element X variant 543cttttcattc caatggggtg gcttc 2554424DNAArtificial sequenceExemplary element X variant 544atacttttca ttcgggtggc ttct 2454524DNAArtificial sequenceExemplary element X variant 545tacttttcat tcgggtggct tctg 2454624DNAArtificial sequenceExemplary element X variant 546acttttcatt cgggtggctt ctgg 2454724DNAArtificial sequenceExemplary element X variant 547cttttcattc gggtggcttc tgga 2454830DNAArtificial sequenceExemplary element X variant 548cttttcattc caatggggtg actttgcttc 3054925DNAArtificial sequenceExemplary element X variant 549cttttcaatg gggtgacttt gcttc 2555025DNAArtificial sequenceExemplary element X variant 550cttttcattc caatggggtg gcttc 2555124DNAArtificial sequenceExemplary element X variant 551atacttttca atggggtggc ttct 2455224DNAArtificial sequenceExemplary element X variant 552tacttttcaa tggggtggct tctg 2455324DNAArtificial sequenceExemplary element X variant 553acttttcaat ggggtggctt ctgg 2455424DNAArtificial sequenceExemplary element X variant 554cttttcaatg gggtggcttc tgga 2455530DNAArtificial sequenceExemplary element X variant 555cttttcattc caatggggtg actttgcttc 3055629DNAArtificial sequenceExemplary element X variant 556atacttttca ttccaatggg gtggcttct 2955729DNAArtificial sequenceExemplary element X variant 557tacttttcat tccaatgggg tggcttctg 2955829DNAArtificial sequenceExemplary element X variant 558acttttcatt ccaatggggt ggcttctgg 2955929DNAArtificial sequenceExemplary element X variant 559cttttcattc caatggggtg gcttctgga 2956030DNAArtificial sequenceExemplary element X variant 560cttttcattc caatggggtg actttgcttc 3056125DNAArtificial sequenceExemplary element X variant 561cttttcattc gggtgacttt gcttc 2556225DNAArtificial sequenceExemplary element X variant 562cttttcattc caatgacttt gcttc 2556324DNAArtificial sequenceExemplary element X variant 563atacttttca ttcactttgc ttct

2456424DNAArtificial sequenceExemplary element X variant 564tacttttcat tcactttgct tctg 2456524DNAArtificial sequenceExemplary element X variant 565acttttcatt cactttgctt ctgg 2456624DNAArtificial sequenceExemplary element X variant 566cttttcattc actttgcttc tgga 2456730DNAArtificial sequenceExemplary element X variant 567cttttcattc caatggggtg actttgcttc 3056825DNAArtificial sequenceExemplary element X variant 568cttttcaatg gggtgacttt gcttc 2556925DNAArtificial sequenceExemplary element X variant 569cttttcattc caatgacttt gcttc 2557024DNAArtificial sequenceExemplary element X variant 570atacttttca atgactttgc ttct 2457124DNAArtificial sequenceExemplary element X variant 571tacttttcaa tgactttgct tctg 2457224DNAArtificial sequenceExemplary element X variant 572acttttcaat gactttgctt ctgg 2457324DNAArtificial sequenceExemplary element X variant 573cttttcaatg actttgcttc tgga 2457430DNAArtificial sequenceExemplary element X variant 574cttttcattc caatggggtg actttgcttc 3057529DNAArtificial sequenceExemplary element X variant 575atacttttca ttccaatgac tttgcttct 2957629DNAArtificial sequenceExemplary element X variant 576tacttttcat tccaatgact ttgcttctg 2957729DNAArtificial sequenceExemplary element X variant 577acttttcatt ccaatgactt tgcttctgg 2957829DNAArtificial sequenceExemplary element X variant 578cttttcattc caatgacttt gcttctgga 2957930DNAArtificial sequenceExemplary element X variant 579cttttcattc caatggggtg actttgcttc 3058029DNAArtificial sequenceExemplary element X variant 580atacttttca ttcgggtgac tttgcttct 2958129DNAArtificial sequenceExemplary element X variant 581tacttttcat tcgggtgact ttgcttctg 2958229DNAArtificial sequenceExemplary element X variant 582acttttcatt cgggtgactt tgcttctgg 2958329DNAArtificial sequenceExemplary element X variant 583cttttcattc gggtgacttt gcttctgga 2958430DNAArtificial sequenceExemplary element X variant 584cttttcattc caatggggtg actttgcttc 3058529DNAArtificial sequenceExemplary element X variant 585atacttttca atggggtgac tttgcttct 2958629DNAArtificial sequenceExemplary element X variant 586tacttttcaa tggggtgact ttgcttctg 2958729DNAArtificial sequenceExemplary element X variant 587acttttcaat ggggtgactt tgcttctgg 2958829DNAArtificial sequenceExemplary element X variant 588cttttcaatg gggtgacttt gcttctgga 2958934DNAArtificial sequenceExemplary element X variant 589atacttttca ttccaatggg gtgactttgc ttct 3459034DNAArtificial sequenceExemplary element X variant 590tacttttcat tccaatgggg tgactttgct tctg 3459134DNAArtificial sequenceExemplary element X variant 591acttttcatt ccaatggggt gactttgctt ctgg 3459234DNAArtificial sequenceExemplary element X variant 592cttttcattc caatggggtg actttgcttc tgga 3459330DNAArtificial sequenceExemplary element X variant 593cttttcattc caatggggtg actttgcttc 3059425DNAArtificial sequenceExemplary element X variant 594cttttcaatg gggtgacttt gcttc 2559525DNAArtificial sequenceExemplary element X variant 595cttttcattc gggtgacttt gcttc 2559624DNAArtificial sequenceExemplary element X variant 596cttttgggtg actttgcttc tgga 2459724DNAArtificial sequenceExemplary element X variant 597acttttgggt gactttgctt ctgg 2459824DNAArtificial sequenceExemplary element X variant 598tacttttggg tgactttgct tctg 2459924DNAArtificial sequenceExemplary element X variant 599atacttttgg gtgactttgc ttct 2460024DNAArtificial sequenceExemplary element X variant 600catacttttg ggtgactttg cttc 2460129DNAArtificial sequenceExemplary element X variant 601gtacttcata gggtgacttt gcttctgga 2960224DNAArtificial sequenceExemplary element X variant 602gtacttcata actttgcttc tgga 2460319DNAArtificial sequenceExemplary element X variant 603gtacttcata gcttctgga 1960434DNAArtificial sequenceExemplary element X variant 604gtacttcata cttttgggtg actttgcttc tgga 3460529DNAArtificial sequenceExemplary element X variant 605gtacttcata cttttacttt gcttctgga 2960624DNAArtificial sequenceExemplary element X variant 606gtacttcata cttttgcttc tgga 2460734DNAArtificial sequenceExemplary element X variant 607gtacttcata cttttcattc actttgcttc tgga 3460829DNAArtificial sequenceExemplary element X variant 608gtacttcata cttttcattc gcttctgga 2960934DNAArtificial sequenceExemplary element X variant 609gtacttcata cttttcattc caatggcttc tgga 3461034DNAArtificial sequenceExemplary element X variant 610gtactctttt caatggggtg actttgcttc tgga 3461129DNAArtificial sequenceExemplary element X variant 611gtactctttt gggtgacttt gcttctgga 2961224DNAArtificial sequenceExemplary element X variant 612gtactctttt actttgcttc tgga 2461319DNAArtificial sequenceExemplary element X variant 613gtactctttt gcttctgga 1961434DNAArtificial sequenceExemplary element X variant 614gtactctttt cattcgggtg actttgcttc tgga 3461529DNAArtificial sequenceExemplary element X variant 615gtactctttt cattcacttt gcttctgga 2961624DNAArtificial sequenceExemplary element X variant 616gtactctttt cattcgcttc tgga 2461734DNAArtificial sequenceExemplary element X variant 617gtactctttt cattccaatg actttgcttc tgga 3461829DNAArtificial sequenceExemplary element X variant 618gtactctttt cattccaatg gcttctgga 2961934DNAArtificial sequenceExemplary element X variant 619gtactctttt cattccaatg gggtggcttc tgga 3462034DNAArtificial sequenceExemplary element X variant 620gtacttcata cattcgggtg actttgcttc tgga 3462129DNAArtificial sequenceExemplary element X variant 621gtacttcata cattcacttt gcttctgga 2962224DNAArtificial sequenceExemplary element X variant 622gtacttcata cattcgcttc tgga 2462334DNAArtificial sequenceExemplary element X variant 623gtacttcata cattccaatg actttgcttc tgga 3462429DNAArtificial sequenceExemplary element X variant 624gtacttcata cattccaatg gcttctgga 2962534DNAArtificial sequenceExemplary element X variant 625gtacttcata cattccaatg gggtggcttc tgga 3462634DNAArtificial sequenceExemplary element X variant 626gtacttcata cttttcaatg actttgcttc tgga 3462729DNAArtificial sequenceExemplary element X variant 627gtacttcata cttttcaatg gcttctgga 2962834DNAArtificial sequenceExemplary element X variant 628gtacttcata cttttcattc gggtggcttc tgga 3462924DNAArtificial sequenceExemplary element X variant 629gtactctttt caatggcttc tgga 2463029DNAArtificial sequenceExemplary element X variant 630gtactctttt caatggggtg gcttctgga 2963129DNAArtificial sequenceExemplary element X variant 631gtactctttt cattcacttt gcttctgga 2963229DNAArtificial sequenceExemplary element X variant 632gtacttcata cattcgggtg gcttctgga 2963344DNAArtificial sequencePPE-1-derived regulatory element scheme 633gtacttcata cttttcattc caatggggtg actttgcttc tgga 4463435DNAArtificial sequencePPE-1-derived regulatory element scheme 634gtacttcata cttttcattc caatggggtg acttt 3563525DNAArtificial sequencePPE-1-derived regulatory element scheme 635cattccaatg gggtgacttt gcttc 2563629DNAArtificial sequencePPE-1-derived regulatory element scheme 636cattccaatg gggtgacttt gcttctgga 2963720DNAArtificial sequencePPE-1-derived regulatory element scheme 637cattccaatg gggtgacttt 2063814DNAArtificial sequencePPE-1-derived regulatory element scheme 638actttgcttc tgga 1463910DNAArtificial sequencePPE-1-derived regulatory element scheme 639actttgcttc 106409DNAArtificial sequencePPE-1-derived regulatory element scheme 640gcttctgga 964114DNAArtificial sequencePPE-1-derived regulatory element scheme 641actttgcttc tgga 1464210DNAArtificial sequencePPE-1-derived regulatory element scheme 642actttgcttc 106439DNAArtificial sequencePPE-1-derived regulatory element scheme 643gcttctgga 964415DNAArtificial sequencePPE-1-derived regulatory element scheme 644cattccaatg gcttc 15

Claims

1. A method of treating a solid tumor in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a non-replicating adenovirus vector, wherein said vector comprises a polynucleotide which comprises a fas-chimera transgene transcriptionally linked to a murine pre-proendothelin promoter, wherein said therapeutically effective amount is at least 1.times.10.sup.8 virus particles.

2. The method of claim 1, wherein said therapeutically effective amount is at least about 1.times.10.sup.9 to about 1.times.10.sup.16 virus particles or at least about 1.times.10.sup.11 to about 1.times.10.sup.13 virus particles.

3. (canceled)

4. The method of claim 1, wherein said therapeutically effective amount is at least about 3.times.10.sup.12 virus particles or at least about 1.times.10.sup.13 virus particles.

5. (canceled)

6. The method of claim 1, wherein said fas-chimera transgene comprises the sequence as set forth in SEQ ID NO: 2.

7. The method of claim 1, wherein said fas-chimera transgene comprises the sequence as set forth in SEQ ID NO: 3.

8. The method of claim 1, wherein said fas-chimera transgene comprises the sequence as set forth in SEQ ID NO: 4.

9. The method of claim 1, wherein said murine pre-pro endothelin promoter comprises an enhancer comprising the sequence as set forth in SEQ ID NO: 6 or the complements sequence thereof.

10. The method of claim 1, wherein said murine pre-pro endothelin promoter comprises an enhancer comprising the sequence as set forth in SEQ ID NO: 8 or the complementary sequence thereof.

11. The method of claim 1, wherein said murine pre-pro endothelin promoter comprises an enhancer comprising the sequence as set forth in SEQ ID NO: 7 or a complementary sequence thereof.

12. The method of claim 1, wherein said murine pre-pro endothelin promoter comprises the sequence as set forth in SEQ ID NO: 13.

13. The method of claim 1, wherein said murine prepro-endothelin promoter comprises the sequence as set forth in SEQ ID NO: 12.

14. (canceled)

15. The method of claim 1, wherein said non-replicating adenovirus vector is an adenovirus 5 vector.

16. The method of claim 1, wherein said adenovirus vector comprises the polynucleotide sequence as set forth in SEQ ID NO: 9 or 10.

17. The method of claim 1, wherein said solid tumor is a cancer.

18. The method of claim 1, wherein said solid tumor is a primary tumor or a metastatic tumor.

19. (canceled)

20. The method of claim 1, wherein said adenovirus vector is administered systemically.

21. The method of claim 20, wherein said vector is administered systemically by a route selected from the group consisting of intra-articular administration, intravenous administration, intraperitoneal administration, subcutaneous administration, infusion, oral administration, rectal administration, nasal administration and inhalation.

22. The method of claim 1, comprising administering said adenovirus vector in at least two separate systemic doses.

23. The method of claim 1, wherein said adenovirus is detected in the blood of said subject at least about 4 days post administration.

24. The method of claim 1, wherein an amount of serum anti-adenovirus antibodies is increased following said administering, and said adenovirus is detected in the blood of said subject at least about 21 days post administration.

25-37. (canceled)

38. The method of claim 1, wherein said non-replicating adenovirus vector is administered to said subject, in at least two separate doses, and wherein said administration does not induce a dose-dependent increase in antibodies against said adenovirus vector in said subject.

39-65. (canceled)

66. The method of claim 1, wherein said administering inhibits angiogenesis of said tumor or inhibits growth of said tumor.

67. (canceled)

68. (canceled)

69. The method of claim 1, wherein said solid tumor is a thyroid cancer.

70. The method of claim 69, wherein said therapeutically effective amount is at least about 1.times.10.sup.13 virus particles.

71. The method of claim 1, wherein said solid tumor is a neuroendocrine cancer.

72. The method of claim 71, wherein said therapeutically effective amount is at least about 1.times.10.sup.13 virus particles.

73. A kit for treating a solid tumor in a subject in need thereof, comprising a unit dosage of virus particles of a non-replicating adenovirus vector comprising the polynucleotide sequence as set forth in SEQ ID NO: 9 or 10 wherein said non-replicating adenovirus vector is formulated for intravenous administration, and instructions for administration of said adenovirus.

74. (canceled)

75. (canceled)

76. A method for administering a therapeutic composition comprising an adenoviral vector to a subject, comprising administering a therapeutically effective amount of the composition to the subject at least twice, wherein the administering does not induce a dose-dependent increase in anti-adenovirus antibodies against said adenoviral vector in the subject.

77-82. (canceled)

83. The method of claim 1, wherein said subject is further receiving a chemotherapeutic agent.

84. The method of claim 83, wherein said chemotherapeutic agent is administered prior to treatment with said virus particles, concomitantly with treatment with said virus particles, or following treatment with said virus particles.

85. (canceled)

86. (canceled)

87. The method of claim 83, wherein said chemotherapeutic agent is sunitinib.

88. An adenovirus vector comprising the polynucleotide sequence as set forth in SEQ ID NO:9.

89. An adenovirus vector comprising the polynucleotide sequence as set forth in SEQ ID NO: 10.

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