Patent application title: ENGINEERED HUMAN-ENDOGENOUS VIRUS-LIKE PARTICLES AND METHODS OF USE THEREOF FOR DELIVERY TO CELLS
Inventors:
J. Keith Joung (Winchester, MA, US)
J. Keith Joung (Winchester, MA, US)
Peter Cabeceiras (Boston, MA, US)
IPC8 Class: AC12N1586FI
USPC Class:
1 1
Class name:
Publication date: 2022-08-18
Patent application number: 20220259617
Abstract:
Human-derived virus-like particles (heVLPs), comprising a membrane
comprising a phospholipid bilayer with one or more HERV-derived envelope
proteins on the external side; one or more HERV-derived GAG proteins in
the heVLP core, and a cargo molecule, e.g., a biomolecule and/or chemical
cargo molecule, disposed in the core of the heVLP on the inside of the
membrane, wherein the heVLP does not comprise a gag protein, except for
gag proteins that are encoded in the human genome or gag proteins that
are encoded by a consensus sequence that is derived from gag proteins
found in the human genome, and methods of use thereof for delivery of the
cargo molecule to cells.Claims:
1. An engineered human-derived virus-like particle (heVLP), comprising: a
membrane comprising a phospholipid bilayer with one or more HERV-derived
ENV/glycoprotein(s) on the external side; a HERV-derived GAG protein in
the heVLP core, and a cargo disposed in the core of the heVLP on the
inside of the membrane, wherein the cargo is fused to a human-endogenous
GAG or other plasma membrane recruitment domain, and the heVLP does not
comprise a non-human gag and/or pol protein.
2. The heVLP of claim 1, wherein the cargo is a therapeutic or diagnostic protein or nucleic acid encoding a therapeutic or diagnostic protein, or a small molecule.
3. The heVLP of claim 1, wherein the cargo is a gene editing reagent.
4. The heVLP of claim 1, wherein the gene editing reagent comprises a zinc finger (ZF), transcription activator-like effector (TALE), and/or CRISPR-based genome editing or modulating protein; a nucleic acid encoding a zinc finger (ZF), transcription activator-like effector (TALE), and/or CRISPR-based genome editing or modulating protein; or a riboucleoprotein complex (RNP) comprising a CRISPR-based genome editing or modulating protein.
5. The heVLP of claim 4, wherein the gene editing reagent is selected from the proteins listed in Tables 2, 3, 4 & 5.
6. The heVLP of claim 4, wherein the gene editing reagent comprises a CRISPR-based genome editing or modulating protein, and the heVLP further comprises one or more guide RNAs that bind to and direct the CRISPR-based genome editing or modulating protein to a target sequence.
7. The heVLP of claim 1, wherein the cargo comprises a fusion to a human-endogenous GAG or other plasma membrane recruitment domain, preferably as shown in Table 6.
8. A method of delivering a cargo molecule to a target cell, optionally a cell in vivo or in vitro, the method comprising contacting the cell with the heVLP of claim 1 comprising the cargo molecule, preferably wherein the cargo molecule is a biomolecule and/or chemical.
9. A method of producing a heVLP comprising one or more cargo molecules, the method comprising: providing a cell expressing one or more HERV-derived envelope proteins, one or more HERV-derived GAG proteins, and the one or more cargo molecules, wherein the cell does not express a gag and/or pol protein, except for gag proteins that are encoded in the human genome or gag proteins that are encoded by a consensus sequence that is derived from gag proteins found in the human genome; and maintaining the cell under conditions such that the cells produce heVLPs.
10. The method of claim 9, further comprising harvesting and optionally purifying and/or concentrating the produced heVLPs.
11. The method of claim 9, wherein the cargo molecule is a therapeutic or diagnostic protein or nucleic acid encoding a therapeutic or diagnostic protein, or a small molecule therapeutic or diagnostic agent.
12. The method of claim 9, wherein the cargo molecule is a gene editing reagent.
13. The method of claim 9, wherein the gene editing reagent comprises a zinc finger (ZF), transcription activator-like effector (TALE), and/or CRISPR-based genome editing or modulating protein; a nucleic acid encoding a zinc finger (ZF), transcription activator-like effector (TALE), and/or CRISPR-based genome editing or modulating protein; or a riboucleoprotein complex (RNP) comprising a CRISPR-based genome editing or modulating protein.
14. The method of claim 13, wherein the gene editing reagent is selected from the proteins listed in Tables 2, 3, 4 & 5.
15. The method of claim 13, wherein the gene editing reagent comprises a CRISPR-based genome editing or modulating protein, and the heVLP further comprises one or more guide RNAs that bind to and direct the CRISPR-based genome editing or modulating protein to a target sequence.
16. The method of claim 9, wherein the cargo molecule comprises a fusion to a human-endogenous GAG or other plasma membrane recruitment domain, preferably as shown in Table 6.
17. A cell that expresses, in combination: one or more HERV-derived envelope proteins; one or more HERV-derived GAG proteins; and a cargo molecule, preferably fused to a human endogenous GAG or other plasma membrane recruitment domain, wherein the cell does not express non-human gag protein.
18. The cell of claim 17, wherein the cargo molecule is a therapeutic or diagnostic protein or nucleic acid encoding a therapeutic or diagnostic protein.
19. The cell of claim 17, wherein the cargo molecule is a gene editing reagent.
20. The cell of claim 17, wherein the gene editing reagent comprises a zinc finger (ZF), transcription activator-like effector (TALE), and/or CRISPR-based genome editing or modulating protein; a nucleic acid encoding a zinc finger (ZF), transcription activator-like effector (TALE), and/or CRISPR-based genome editing or modulating protein; or a riboucleoprotein complex (RNP) comprising a CRISPR-based genome editing or modulating protein.
21. The cell of claim 20, wherein the gene editing reagent is selected from the proteins listed in Tables 2, 3, 4 & 5.
22. The cell of claim 20, wherein the gene editing reagent comprises a CRISPR-based genome editing or modulating protein, and the heVLP further comprises one or more guide RNAs that bind to and direct the CRISPR-based genome editing or modulating protein to a target sequence.
23. The cells of claim 17, wherein the cargo molecule comprises a fusion to a human-endogenous GAG or other plasma membrane recruitment domain, preferably as shown in Table 6.
24. The cells of claim 17, wherein the cells are primary or stable human cell lines.
25. The cells of claim 24, which are Human Embryonic Kidney (HEK) 293 cells, HEK293 T cells, or BeWo cells.
Description:
CLAIM OF PRIORITY
[0001] This application claims the benefit of U.S. Patent Application Ser. No. 62/861,186, filed on Jun. 13, 2019. The entire contents of the foregoing are hereby incorporated by reference.
TECHNICAL FIELD
[0003] Described herein are engineered human-endogenous virus-like particles (heVLPs) comprising a membrane comprising a phospholipid bilayer on the external side; and a cargo, e.g., a biomolecule and/or chemical cargo, disposed in the core of the heVLP on the inside of the membrane, wherein the heVLP does not comprise a protein from non-human gag or pol, and methods of use thereof for delivery of the cargo to cells.
BACKGROUND
[0004] Delivery of cargo such as proteins, nucleic acids, and/or chemicals into the cytosol of living cells has been a significant hurdle in the development of biological therapeutics.
SUMMARY
[0005] Described herein are heVLPs that are capable of packaging and delivering DNA, RNA, protein, chemical compounds and/or molecules, and any combination of these four entities into eukaryotic cells. The non-viral heVLP systems described herein have the potential to be simpler, more efficient and safer than conventional, artificially-derived lipid/gold nanoparticles and viral particle-based delivery systems because heVLPs are comprised of human-derived components. The cargo inside may or may not be human derived, but the heVLP is entirely comprised from human and synthetic non-immunogenic components. "Synthetic" components include surface scFv/nanobody/darpin peptides that have been demonstrated to not be immunostimulatory and can be used to enhance targeting and cellular uptake of heVLPs. This means that the exterior surface of the particle lacks components that are significantly immunostimulatory, which should minimize immunogenicity and antibody neutralization of these particles. Excluding cargo, the heVLPs do not contain exogenous viral components inherent to other VLPs and this represents a significant and novel advancement in technology. In addition, heVLPs can utilize (but do not require) chemical-based dimerizers, and heVLPs have the ability to package and deliver cargo molecules including therapeutic or diagnostic agents, including biomolecules and chemicals, e.g., specialty single and/or double-stranded DNA molecules (e.g., plasmid, mini circle, closed-ended linear DNA, AAV DNA, episomes, bacteriophage DNA, homology directed repair templates, etc.), single and/or double-stranded RNA molecules (e.g., single guide RNA, prime editing guide RNA, messenger RNA, transfer RNA, long non-coding RNA, circular RNA, RNA replicon, circular or linear splicing RNA, micro RNA, small interfering RNA, short hairpin RNA, piwi-interacting RNA, toehold switch RNA, RNAs that can be bound by RNA binding proteins, bacteriophage RNA, internal ribosomal entry site containing RNA, etc.), proteins, chemical compounds and/or molecules (e.g., small molecules), and combinations of the above listed cargos (e.g. AAV particles).
[0006] The heVLPs described herein are different from conventional retroviral particles, virus-like particles (VLPs), exosomes and other previously described extracellular vesicles that can be loaded with cargo, at least because heVLPs can be produced by a strategic overexpression of human-derived components in human cells, heVLPs have a vast diversity of possible cargos and loading strategies, heVLPs lack a limiting DNA/RNA length constraint, heVLPs lack proteins derived from pol and exogenous gag, and heVLPs have unique mechanisms of cellular entry.
[0007] Described herein are compositions and methods for cargo delivery that can be used with a diverse array of protein and nucleic acid molecules, including genome editing, epigenome modulation, transcriptome editing and proteome modulation reagents, that are applicable to many disease therapies.
[0008] Thus, provided herein are engineered heVLPs, comprising a membrane comprising a phospholipid bilayer with one or more HERV-derived ENV/glycoprotein(s) (e.g., overexpressed from exogenous sources, such as plasmids or stably integrated transgenes, in heVLP production cells) (e.g., as shown in Table 1) on the external side; and a human endogenous GAG protein, other plasma membrane recruitment domain, and/or biomolecule/chemical cargo disposed in the core of the heVLP on the inside of the membrane, wherein the biomolecule cargo may or may not be fused to a human-endogenous GAG or other plasma membrane recruitment domain (e.g., as shown in Table 6), and the heVLP does not comprise a non-human gag and/or pol protein, do not express gag and/or pol proteins except for gag proteins that are encoded in the human genome or gag proteins that are encoded by a consensus sequence that is derived from gag proteins found in the human genome. Human-derived GAG or other plasma membrane recruitment domains fused to biomolecule cargo can be overexpressed from exogenous sources, such as plasmids or stably integrated transgenes, in heVLP production cells.
[0009] In some embodiments, the HERV ENV can be truncated or fused to an scFv or other targeting polypeptides.
[0010] In some embodiments the HERV GAG can be fused to a plasma membrane recruitment domain (e.g., as shown in Table 6).
[0011] In another embodiment, engineered heVLPs comprise a membrane comprising a phospholipid bilayer with one or more HERV-derived ENV/glycoprotein(s) (e.g., overexpressed from exogenous sources, such as plasmids or stably integrated transgenes, in heVLP production cells) (e.g., as shown in Table 1) on the external side; and, if desired, a plasma membrane recruitment domain (e.g., as shown in Table 6); and, if desired a biomolecule/chemical cargo inside the particle.
[0012] Also provided are methods of delivering a cargo to a target cell, e.g., a cell in vivo or in vitro, by contacting the cell with the heVLP of claim 1 comprising the biomolecule and/or chemical as cargo.
[0013] In addition, provided herein are methods for producing a heVLP comprising a biomolecular cargo. The methods include providing a cell expressing (e.g., engineered to express or overexpress) one or more HERV-derived envelope proteins (e.g., as shown in Table 1), and a cargo, wherein the cell does not express a gag and/or pol protein, except for gag proteins that are encoded in the human genome or gag proteins that are encoded by a consensus sequence that is derived from gag proteins found in the human genome; and maintaining the cell under conditions such that the cells produce heVLPs. In some embodiments, the methods further include harvesting and optionally purifying and/or concentrating the produced heVLPs.
[0014] Also provided herein are cells (e.g., isolated cells, preferably mammalian, e.g., human, cells) that express, e.g., that have been induced to overexpress, in combination one or more HERV-derived envelope proteins (e.g., (overexpressed from exogenous sources, such as plasmids or stably integrated transgenes)(e.g., as shown in Table 1), and a cargo fused to a human endogenous GAG or other plasma membrane recruitment domain (e.g., as shown in Table 6), wherein the cell does not express a gag protein except for gag proteins that are encoded in the human genome or gag proteins that are encoded by a consensus sequence that is derived from gag proteins found in the human genome (overexpressed from exogenous sources, such as plasmids or stably integrated transgenes)). In some embodiments, the cells are primary or stable human cell lines, e.g., Human Embryonic Kidney (HEK) 293 cells, HEK293 T cells, or BeWo cells. The cells can be used to produce heVLPs as described herein.
[0015] In some embodiments, the methods include using cells that have or have not been manipulated to express any exogenous proteins except for a HERV envelope (e.g., as shown in Table 1), and, if desired, a plasma membrane recruitment domain (e.g., as shown in Table 6). In this embodiment, the "empty" particles that are produced can be loaded with biomolecule or chemical molecule cargo by utilizing nucleofection, lipid, polymer, or CaCl.sub.2 transfection, sonication, freeze thaw, and/or heat shock of purified particles mixed with cargo. In all embodiments, producer cells do not express any human exogenous gag protein. This type of loading allows for cargo to be unmodified by fusions to plasma membrane recruitment domains and represents a significant advancement from previous VLP technology.
[0016] In another embodiment, heVLPs that contain cargo are produced and isolated can be loaded with additional biomolecule or chemical molecule cargo by utilizing nucleofection, lipid, polymer, or CaCl.sub.2 transfection, sonication, freeze thaw, incubation at various temperatures, and/or heat shock of purified particles mixed with cargo.
[0017] In some embodiments, the cargo is a therapeutic or diagnostic protein or nucleic acid encoding a therapeutic or diagnostic protein.
[0018] In some embodiments, the cargo is a chemical compound or molecule.
[0019] In some embodiments, the chemical molecule is a trigger for protein-protein dimerization of multimerization, such as the A/C heterodimerizer or rapamycin.
[0020] In some embodiments, the chemical compound is a DNA PK inhibitor, such as M3814, NU7026, or NU7441 which potently enhance homology directed repair gene editing.
[0021] In some embodiments, the biomolecule cargo is a gene editing reagent.
[0022] In some embodiments, the gene editing reagent comprises a zinc finger (ZF), transcription activator-like effector (TALE), and/or CRISPR-based genome editing or modulating protein; a nucleic acid encoding a zinc finger (ZF), transcription activator-like effector (TALE), and/or CRISPR-based genome editing or modulating protein; or a riboucleoprotein complex (RNP) comprising a CRISPR-based genome editing or modulating protein.
[0023] In some embodiments, the gene editing reagent is selected from the proteins listed in Tables 2, 3, 4 & 5.
[0024] In some embodiments, the gene editing reagent comprises a CRISPR-based genome editing or modulating protein, and the heVLP further comprises one or more guide RNAs that bind to and direct the CRISPR-based genome editing or modulating protein to a target sequence.
[0025] In some embodiments, the cargo comprises a covalent or non-covalent connection to a human-endogenous GAG or other plasma membrane recruitment domain, preferably as shown in Table 6. Covalent connections, for example, can include direct protein-protein fusions generated from a single reading frame, inteins that can form peptide bonds, other proteins that can form covalent connections at R-groups and/or RNA splicing. Non-covalent connections, for example, can include DNA/DNA, DNA/RNA, and/or RNA/RNA hybrids (nucleic acids base pairing to other nucleic acids via hydrogen-bonding interactions), protein domains that dimerize or multimerize with or without the need for a chemical compound/molecule to induce the protein-protein binding, single chain variable fragments, nanobodies, affibodies, proteins that bind to DNA and/or RNA, proteins with quaternary structural interactions, optogenetic protein domains that can dimerize or multimerize in the presence of certain light wavelengths, and/or naturally reconstituting split proteins.
[0026] In some embodiments, the cargo comprises a fusion to a dimerization domain or protein-protein binding domain that may or may not require a molecule to trigger dimerization or protein-protein binding.
[0027] In some embodiments, the producer cells are FDA-approved cells lines, allogenic cells, and/or autologous cells derived from a donor.
[0028] In some embodiments, the full or active peptide domains of human CD47 may be incorporated in the heVLP surface to reduce immunogenicity.
[0029] Examples of AAV proteins included here are AAV REP 52, REP 78, and VP1-3. The capsid site where proteins can be inserted is T138 starting from the VP1 amino acid counting. Dimerization domains could be inserted at this point in the capsid, for instance.
[0030] Examples of dimerization domains included here that may or may not need a small molecule inducer are dDZF1, dDZF2, DmrA, DmrB, DmrC, FKBP, FRB, GCN4 scFv, 10.times./24.times. GCN4, GFP nanobody and GFP.
[0031] Examples of split inteins included here are Npu DnaE, Cfa, Vma, and Ssp DnaE.
[0032] Examples of other split proteins included here that make a covalent bond together are Spy Tag and Spy Catcher.
[0033] Examples of RNA binding proteins included here are MS2, Com, and PP7.
[0034] Examples of synthetic DNA-binding zinc fingers included here are ZF6/10, ZF8/7, ZF9, MK10, Zinc Finger 268, and Zinc Finger 268/NRE.
[0035] Examples of proteins that multimerize as a result of quaternary structure included here are E. coli ferritin, and the other chimeric forms of ferritin.
[0036] Examples of optogenetic "light-inducible proteins" included here are Cry2, CIBN, and Lov2-Ja.
[0037] Examples of peptides the enhance transduction included here are L17E, Vectofusin, KALA, and the various forms of nisin.
[0038] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Methods and materials are described herein for use in the present invention; other, suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.
[0039] Other features and advantages of the invention will be apparent from the following detailed description and figures, and from the claims.
DESCRIPTION OF DRAWINGS
[0040] FIG. 1: Depiction of exemplary T2heVLP/T4heVLP production and transduction for RNP/protein delivery. All heVLP expression constructs are stably integrated in the genome of the producer cell. Construct 1-0 corresponds to the human-endogenous GAG (hGAG). Construct 1-1 corresponds to the human-endogenous GAG or other phospholipid bilayer recruitment domain. 1-2 corresponds to the cargo. 2 corresponds to an optional guide RNA. 1-0, 1-1 and 1-2 are translated in the cytosol where the fusion of 1-1 and 1-2 complexes with guide RNA before it is recruited to the phospholipid bilayer. 3 corresponds to a HERV-derived glycoprotein (hENV). The HERV-derived glycoprotein is expressed as a transmembrane protein on the plasma membrane. hGAG drives budding of cargo-containing heVLPs from the plasma membrane to extracellular space. These particles are purified and are able to fuse with target cells and deliver cargo by interacting with surface receptors at the target cell surface.
[0041] FIG. 2: Depiction of purified heVLPs entering a target cell and delivering cargo to the cytosol. Importantly, the human-endogenous GAG or other phospholipid bilayer recruitment domain allows cargo to enter the target cell nucleus as long as cargo possesses a nuclear localization sequence.
[0042] FIG. 3: Exemplary T1heVLP-delivered spCas9 genome editing in vitro. HEK 293T cells transduced with T1heVLPs containing PLC PH fused to spCas9, hGAGK.sub.con fused to spCas9, or human Activity-regulated cytoskeleton-associated protein (hArc) fused to spCas9 targeted to VEGF site #3. heVLPs are pseudotyped with either hENVW (left chart) or hENVFRD (right chart). Gene modification is measured by amplicon sequencing.
[0043] FIG. 4: Depiction of T1heVLP/T3heVLP production. Plasmid DNA constructs involved in the transfection encode cargo, an optional guide RNA, hGAG and a HERV-derived glycoprotein. Plasmids, or other types of DNA molecules, will be distributed throughout the production cell, so constructs located in the nucleus will express heVLP components and cargo, and constructs located near the plasma membrane or endosomes will be encapsulated within budding heVLPs.
[0044] FIG. 5: Depiction of exemplary heVLP and cargo configuration
[0045] This particle was created by producer cells expressing an envelope protein. Cargo was packaged inside the particle either by producer cells expressing cargo or particles being loaded by various particle loading methods described herein, such as electroporation.
[0046] FIG. 6: Depiction of exemplary heVLP and cargo configuration
[0047] This particle was created by producer cells expressing an envelope protein. Cargo was packaged inside the particle either by producer cells expressing cargo-gag fusion or particles being loaded by various particle loading methods described herein, such as electroporation.
[0048] FIG. 7: Depiction of exemplary heVLP and cargo configuration
[0049] This particle was created by producer cells expressing an envelope protein. Cargo was packaged inside the particle either by producer cells expressing cargo-PH fusion or particles being loaded by various particle loading methods described herein, such as electroporation.
[0050] FIG. 8: Depiction of exemplary heVLP and cargo configuration
[0051] This particle was created by producer cells expressing an envelope protein. Cargo was packaged inside the particle either by producer cells expressing cargo-gag/PH fusion or particles being loaded by various particle loading methods described herein, such as electroporation.
[0052] FIG. 9: Depiction of exemplary heVLP and cargo configuration
[0053] This particle was created by producer cells expressing an envelope protein. Cargo was packaged inside the particle in the presence of a dimerization molecule (A/C heterodimerizer) either by producer cells expressing cargo and gag fused to DmrA or DmrC or particles being loaded by various particle loading methods described herein, such as electroporation.
[0054] FIG. 10: Depiction of exemplary heVLP and cargo configuration
[0055] This particle was created by producer cells expressing an envelope protein. Cargo was packaged inside the particle in the presence of a dimerization molecule (A/C heterodimerizer) either by producer cells expressing cargo and PH fused to DmrA or DmrC or particles being loaded by various particle loading methods described herein, such as electroporation.
[0056] FIG. 11: Depiction of exemplary heVLP and cargo configuration
[0057] This particle was created by producer cells expressing an envelope protein. Cargo was packaged inside the particle in the presence of a dimerization molecule (A/C heterodimerizer) either by producer cells expressing cargo and gag/PH fused to DmrA or DmrC or particles being loaded by various particle loading methods described herein, such as electroporation.
[0058] FIG. 12: Depiction of exemplary heVLP and cargo configuration
[0059] This particle was created by producer cells expressing an envelope protein. Cargo was packaged inside the particle either by producer cells expressing cargo and gag fused to an RNA binding protein (RBP), MS2, that binds to its MS2 RNA stem loop (MS2 SL) that is complexed with cargo or particles being loaded by various particle loading methods described herein, such as electroporation.
[0060] FIG. 13: Depiction of exemplary heVLP and cargo configuration
[0061] This particle was created by producer cells expressing an envelope protein. Cargo was packaged inside the particle either by producer cells expressing cargo and PH fused to an RNA binding protein (RBP), MS2, that binds to its RNA stem loop (MS2 SL) that is complexed with cargo or particles being loaded by various particle loading methods described herein, such as electroporation.
[0062] FIG. 14: Depiction of exemplary heVLP and cargo configuration
[0063] This particle was created by producer cells expressing an envelope protein. Cargo was packaged inside the particle either by producer cells expressing cargo and gag/PH fused to an RNA binding protein (RBP), MS2, that binds to its RNA stem loop (MS2 SL) that is complexed with cargo or particles being loaded by various particle loading methods described herein, such as electroporation.
[0064] FIG. 15: Depiction of exemplary heVLP and cargo configuration
[0065] This particle was created by producer cells expressing an envelope protein. Cargo was packaged inside the particle in the presence of dimerization molecule (A/C Heterodimerizer) either by producer cells expressing cargo and gag and an RNA binding protein (RBP), MS2, fused to DmrA or DmrC that binds to its RNA stem loop (MS2 SL) that is complexed with cargo or particles being loaded by various particle loading methods described herein, such as electroporation.
[0066] FIG. 16: Depiction of exemplary heVLP and cargo configuration
[0067] This particle was created by producer cells expressing an envelope protein. Cargo was packaged inside the particle in the presence of dimerization molecule (A/C Heterodimerizer) either by producer cells expressing cargo and PH and an RNA binding protein (RBP), MS2, fused to DmrA or DmrC that binds to its RNA stem loop (MS2 SL) that is complexed with cargo or particles being loaded by various particle loading methods described herein, such as electroporation.
[0068] FIG. 17: Depiction of exemplary heVLP and cargo configuration
[0069] This particle was created by producer cells expressing an envelope protein. Cargo was packaged inside the particle in the presence of dimerization molecule (A/C Heterodimerizer) either by producer cells expressing cargo and gag/PH and an RNA binding protein (RBP), MS2, fused to DmrA or DmrC that binds to its RNA stem loop (MS2 SL) that is complexed with cargo or particles being loaded by various particle loading methods described herein, such as electroporation.
[0070] FIG. 18: Depiction of exemplary heVLP and cargo configuration
[0071] This particle was created by producer cells expressing an envelope protein. Cargo was packaged inside the particle either by producer cells expressing cargo and gag fused to a repetitive GCN4 domain that is bound by an scFv that is fused with cargo or particles being loaded by various particle loading methods described herein, such as electroporation.
[0072] FIG. 19: Depiction of exemplary heVLP and cargo configuration
[0073] This particle was created by producer cells expressing an envelope protein. Cargo was packaged inside the particle either by producer cells expressing cargo and PH fused to a repetitive GCN4 domain that is bound by an scFv that is fused with cargo or particles being loaded by various particle loading methods described herein, such as electroporation.
[0074] FIG. 20: Depiction of exemplary heVLP and cargo configuration
[0075] This particle was created by producer cells expressing an envelope protein. Cargo was packaged inside the particle either by producer cells expressing cargo and gag/PH fused to a repetitive GCN4 domain that is bound by an scFv that is fused with cargo or particles being loaded by various particle loading methods described herein, such as electroporation.
[0076] FIG. 21: Depiction of exemplary heVLP and cargo configuration
[0077] This particle was created by producer cells expressing an envelope protein. Cargo was packaged inside the particle in the presence of a dimerization molecule (A/C Heterodimerizer) by producer cells expressing gag and a repetitive GCN4 domain that are fused to DmrA or DmrC. GCN4 is bound by an scFv that is fused with cargo that is also being expressed in producer cells. Particles could also be loaded by various particle loading methods described herein, such as electroporation.
[0078] FIG. 22: Depiction of exemplary heVLP and cargo configuration
[0079] This particle was created by producer cells expressing an envelope protein. Cargo was packaged inside the particle in the presence of a dimerization molecule (A/C Heterodimerizer) by producer cells expressing PH and a repetitive GCN4 domain that are fused to DmrA or DmrC. GCN4 is bound by an scFv that is fused with cargo that is also being expressed in producer cells. Particles could also be loaded by various particle loading methods described herein, such as electroporation.
[0080] FIG. 23: Depiction of exemplary heVLP and cargo configuration
[0081] This particle was created by producer cells expressing an envelope protein. Cargo was packaged inside the particle in the presence of a dimerization molecule (A/C Heterodimerizer) by producer cells expressing gag/PH and a repetitive GCN4 domain that are fused to DmrA or DmrC. GCN4 is bound by an scFv that is fused with cargo that is also being expressed in producer cells. Particles could also be loaded by various particle loading methods described herein, such as electroporation.
[0082] FIG. 24: Depiction of exemplary heVLP and cargo configuration
[0083] This particle was created by producer cells expressing an envelope protein. Cargo (AAV particles) was packaged inside the particle either by producer cells expressing cargo or particles being loaded by various particle loading methods described herein, such as electroporation.
[0084] FIG. 25: Depiction of exemplary heVLP and cargo configuration
[0085] This particle was created by producer cells expressing an envelope protein. Cargo (AAV particles) was packaged inside the particle either by producer cells expressing cargo and gag or particles being loaded by various particle loading methods described herein, such as electroporation.
[0086] FIG. 26: Depiction of exemplary heVLP and cargo configuration
[0087] This particle was created by producer cells expressing an envelope protein. Cargo (AAV particles) was packaged inside the particle either by producer cells expressing cargo and PH or particles being loaded by various particle loading methods described herein, such as electroporation.
[0088] FIG. 27: Depiction of exemplary heVLP and cargo configuration
[0089] This particle was created by producer cells expressing an envelope protein. Cargo (AAV particles) was packaged inside the particle either by producer cells expressing cargo and gag/PH or particles being loaded by various particle loading methods described herein, such as electroporation.
[0090] FIG. 28: Depiction of exemplary heVLP and cargo configuration
[0091] This particle was created by producer cells expressing an envelope protein. Cargo (AAV particles with DmrB inserted in the Capsid protein, VP2) was packaged inside the particle in the presence of DmrB dimerizer molecule either by producer cells expressing cargo and gag fused to DmrB or particles being loaded by various particle loading methods described herein, such as electroporation.
[0092] FIG. 29: Depiction of exemplary heVLP and cargo configuration
[0093] This particle was created by producer cells expressing an envelope protein. Cargo (AAV particles with DmrB inserted in the Capsid protein, VP2) was packaged inside the particle in the presence of DmrB dimerizer molecule either by producer cells expressing cargo and PH fused to DmrB or particles being loaded by various particle loading methods described herein, such as electroporation.
[0094] FIG. 30: Depiction of exemplary heVLP and cargo configuration
[0095] This particle was created by producer cells expressing an envelope protein. Cargo (AAV particles with DmrB inserted in the Capsid protein, VP2) was packaged inside the particle in the presence of DmrB dimerizer molecule either by producer cells expressing cargo and gag/PH fused to DmrB or particles being loaded by various particle loading methods described herein, such as electroporation.
[0096] FIG. 31: Depiction of exemplary heVLP and cargo configuration
[0097] This particle was created by producer cells expressing an envelope protein. Cargo (AAV particles with DmrB inserted in the Capsid protein, VP2) was packaged inside the particle in the presence of DmrB dimerizer and A/C Heterodimerizer molecules either by producer cells expressing cargo and gag fused to DmrA, DmrB, or DmrC, or particles being loaded by various particle loading methods described herein, such as electroporation.
[0098] FIG. 32: Depiction of exemplary heVLP and cargo configuration
[0099] This particle was created by producer cells expressing an envelope protein. Cargo (AAV particles with DmrB inserted in the Capsid protein, VP2) was packaged inside the particle in the presence of DmrB dimerizer and A/C Heterodimerizer molecules either by producer cells expressing cargo and PH fused to DmrA, DmrB, or DmrC, or particles being loaded by various particle loading methods described herein, such as electroporation.
[0100] FIG. 33: Depiction of exemplary heVLP and cargo configuration
[0101] This particle was created by producer cells expressing an envelope protein. Cargo (AAV particles with DmrB inserted in the Capsid protein, VP2) was packaged inside the particle in the presence of DmrB dimerizer and A/C Heterodimerizer molecules either by producer cells expressing cargo and gag/PH fused to DmrA, DmrB, or DmrC, or particles being loaded by various particle loading methods described herein, such as electroporation.
[0102] FIG. 34: Depiction of exemplary heVLP and cargo configuration
[0103] This particle was created by producer cells expressing an envelope protein. Cargo (single-stranded DNA) can be packaged inside the particle by various particle loading methods described herein, such as electroporation.
[0104] FIG. 35: Depiction of exemplary heVLP and cargo configuration
[0105] This particle was created by producer cells expressing an envelope protein and gag. Cargo (single-stranded DNA) can be packaged inside the particle by various particle loading methods described herein, such as electroporation.
[0106] FIG. 36: Depiction of exemplary heVLP and cargo configuration
[0107] This particle was created by producer cells expressing an envelope protein and PH. Cargo (single-stranded DNA) can be packaged inside the particle by various particle loading methods described herein, such as electroporation.
[0108] FIG. 37: Depiction of exemplary heVLP and cargo configuration
[0109] This particle was created by producer cells expressing an envelope protein and gag/PH. Cargo (single-stranded DNA) can be packaged inside the particle by various particle loading methods described herein, such as electroporation.
[0110] FIG. 38: Depiction of exemplary heVLP and cargo configuration
[0111] This particle was created by producer cells expressing an envelope protein. Cargo (double-stranded DNA) can be packaged inside the particle by various particle loading methods described herein, such as electroporation.
[0112] FIG. 39: Depiction of exemplary heVLP and cargo configuration
[0113] This particle was created by producer cells expressing an envelope protein and gag. Cargo (double-stranded DNA) can be packaged inside the particle by various particle loading methods described herein, such as electroporation.
[0114] FIG. 40: Depiction of exemplary heVLP and cargo configuration
[0115] This particle was created by producer cells expressing an envelope protein and PH. Cargo (double-stranded DNA) can be packaged inside the particle by various particle loading methods described herein, such as electroporation.
[0116] FIG. 41: Depiction of exemplary heVLP and cargo configuration
[0117] This particle was created by producer cells expressing an envelope protein and gag/PH. Cargo (double-stranded DNA) can be packaged inside the particle by various particle loading methods described herein, such as electroporation.
[0118] FIG. 42: Depiction of exemplary heVLP and cargo configuration
[0119] This particle was created by producer cells expressing an envelope protein and gag fused to a zinc finger protein (ZFP) that will bind a specific sequence in the cargo. Cargo (double-stranded DNA) can be packaged inside the particle by various particle loading methods described herein, such as electroporation.
[0120] FIG. 43: Depiction of exemplary heVLP and cargo configuration
[0121] This particle was created by producer cells expressing an envelope protein and PH fused to a zinc finger protein (ZFP) that will bind a specific sequence in the cargo. Cargo (double-stranded DNA) can be packaged inside the particle by various particle loading methods described herein, such as electroporation.
[0122] FIG. 44: Depiction of exemplary heVLP and cargo configuration
[0123] This particle was created by producer cells expressing an envelope protein and gag/PH fused to a zinc finger protein (ZFP) that will bind a specific sequence in the cargo. Cargo (double-stranded DNA) can be packaged inside the particle by various particle loading methods described herein, such as electroporation.
[0124] FIG. 45: Depiction of exemplary heVLP and cargo configuration
[0125] This particle was created by producer cells expressing an envelope protein and gag and a zinc finger protein (ZFP) that will bind a specific sequence in the cargo fused to DmrA or DmrC in the presence of A/C Heterodimerizer molecule. Cargo (double-stranded DNA) can be packaged inside the particle by various particle loading methods described herein, such as electroporation.
[0126] FIG. 46: Depiction of exemplary heVLP and cargo configuration
[0127] This particle was created by producer cells expressing an envelope protein and PH and a zinc finger protein (ZFP) that will bind a specific sequence in the cargo fused to DmrA or DmrC in the presence of A/C Heterodimerizer molecule. Cargo (double-stranded DNA) can be packaged inside the particle by various particle loading methods described herein, such as electroporation.
[0128] FIG. 47: Depiction of exemplary heVLP and cargo configuration
[0129] This particle was created by producer cells expressing an envelope protein and gag/PH and a zinc finger protein (ZFP) that will bind a specific sequence in the cargo fused to DmrA or DmrC in the presence of A/C Heterodimerizer molecule. Cargo (double-stranded DNA) can be packaged inside the particle by various particle loading methods described herein, such as electroporation.
[0130] FIG. 48: Depiction of exemplary heVLP and cargo configuration
[0131] This particle was created by producer cells expressing an envelope protein and gag fused to a zinc finger protein (ZFP) that will bind a specific sequence in the cargo. Cargo (double-stranded DNA bound by Cas9 RNP-ZFP fusion) can be packaged inside the particle by various particle loading methods described herein, such as electroporation. Alternatively, the Cas9 RNP-ZFP fusion could be expressed by the producer cells and the particles could be loaded by various particle loading methods described herein, such as electroporation.
[0132] FIG. 49: Depiction of exemplary heVLP and cargo configuration
[0133] This particle was created by producer cells expressing an envelope protein and PH fused to a zinc finger protein (ZFP) that will bind a specific sequence in the cargo. Cargo (double-stranded DNA bound by Cas9 RNP-ZFP fusion) can be packaged inside the particle by various particle loading methods described herein, such as electroporation. Alternatively, the Cas9 RNP-ZFP fusion could be expressed by the producer cells and the particles could be loaded by various particle loading methods described herein, such as electroporation.
[0134] FIG. 50: Depiction of exemplary heVLP and cargo configuration
[0135] This particle was created by producer cells expressing an envelope protein and gag/PH fused to a zinc finger protein (ZFP) that will bind a specific sequence in the cargo. Cargo (double-stranded DNA bound by Cas9 RNP-ZFP fusion) can be packaged inside the particle by various particle loading methods described herein, such as electroporation. Alternatively, the Cas9 RNP-ZFP fusion could be expressed by the producer cells and the particles could be loaded by various particle loading methods described herein, such as electroporation.
[0136] FIG. 51: Depiction of exemplary heVLP and cargo configuration
[0137] This particle was created by producer cells expressing an envelope protein and gag fused to a zinc finger protein (ZFP) fused to DmrA or DmrC that will bind a specific sequence in the cargo in the presence of A/C Heterodimerizer molecule. Cargo (double-stranded DNA bound by Cas9 RNP-ZFP fusion) can be packaged inside the particle by various particle loading methods described herein, such as electroporation. Alternatively, the Cas9 RNP-ZFP fusion could be expressed by the producer cells and the particles could be loaded by various particle loading methods described herein, such as electroporation.
[0138] FIG. 52: Depiction of exemplary heVLP and cargo configuration
[0139] This particle was created by producer cells expressing an envelope protein and PH fused to a zinc finger protein (ZFP) fused to DmrA or DmrC that will bind a specific sequence in the cargo in the presence of A/C Heterodimerizer molecule. Cargo (double-stranded DNA bound by Cas9 RNP-ZFP fusion) can be packaged inside the particle by various particle loading methods described herein, such as electroporation. Alternatively, the Cas9 RNP-ZFP fusion could be expressed by the producer cells and the particles could be loaded by various particle loading methods described herein, such as electroporation.
[0140] FIG. 53: Depiction of exemplary heVLP and cargo configuration
[0141] This particle was created by producer cells expressing an envelope protein and gag/PH fused to a zinc finger protein (ZFP) fused to DmrA or DmrC that will bind a specific sequence in the cargo in the presence of A/C Heterodimerizer molecule. Cargo (double-stranded DNA bound by Cas9 RNP-ZFP fusion) can be packaged inside the particle by various particle loading methods described herein, such as electroporation. Alternatively, the Cas9 RNP-ZFP fusion could be expressed by the producer cells and the particles could be loaded by various particle loading methods described herein, such as electroporation.
[0142] FIG. 54: Depiction of exemplary heVLP and cargo configuration
[0143] This particle was created by producer cells expressing an envelope protein. Cargo (RNA) was packaged inside the particle either by producer cells expressing cargo or particles being loaded by various particle loading methods described herein, such as electroporation.
[0144] FIG. 55: Depiction of exemplary heVLP and cargo configuration
[0145] This particle was created by producer cells expressing an envelope protein. Cargo (RNA) was packaged inside the particle either by producer cells expressing cargo and gag or particles being loaded by various particle loading methods described herein, such as electroporation.
[0146] FIG. 56: Depiction of exemplary heVLP and cargo configuration
[0147] This particle was created by producer cells expressing an envelope protein. Cargo (RNA) was packaged inside the particle either by producer cells expressing cargo and PH or particles being loaded by various particle loading methods described herein, such as electroporation.
[0148] FIG. 57: Depiction of exemplary heVLP and cargo configuration
[0149] This particle was created by producer cells expressing an envelope protein. Cargo (RNA) was packaged inside the particle either by producer cells expressing cargo and gag/PH or particles being loaded by various particle loading methods described herein, such as electroporation.
[0150] FIG. 58: Depiction of exemplary heVLP and cargo configuration
[0151] This particle was created by producer cells expressing an envelope protein. Cargo (RNA with MS2 stem loop(s)) was packaged inside the particle either by producer cells expressing cargo and gag fused to MS2 or particles being loaded by various particle loading methods described herein, such as electroporation.
[0152] FIG. 59: Depiction of exemplary heVLP and cargo configuration
[0153] This particle was created by producer cells expressing an envelope protein. Cargo (RNA with MS2 stem loop(s)) was packaged inside the particle either by producer cells expressing cargo and PH fused to MS2 or particles being loaded by various particle loading methods described herein, such as electroporation.
[0154] FIG. 60: Depiction of exemplary heVLP and cargo configuration
[0155] This particle was created by producer cells expressing an envelope protein. Cargo (RNA with MS2 stem loop(s)) was packaged inside the particle either by producer cells expressing cargo and gag/PH fused to MS2 or particles being loaded by various particle loading methods described herein, such as electroporation.
[0156] FIG. 61: Depiction of exemplary heVLP and cargo configuration
[0157] This particle was created by producer cells expressing an envelope protein. Cargo (RNA with MS2 stem loop(s)) was packaged inside the particle either by producer cells expressing cargo and gag and MS2 fused to DmrA or DmrC in the presence of A/C heterodimerizer, or particles being loaded by various particle loading methods described herein, such as electroporation.
[0158] FIG. 62: Depiction of exemplary heVLP and cargo configuration
[0159] This particle was created by producer cells expressing an envelope protein. Cargo (RNA with MS2 stem loop(s)) was packaged inside the particle either by producer cells expressing cargo and PH and MS2 fused to DmrA or DmrC in the presence of A/C heterodimerizer, or particles being loaded by various particle loading methods described herein, such as electroporation.
[0160] FIG. 63: Depiction of exemplary heVLP and cargo configuration
[0161] This particle was created by producer cells expressing an envelope protein. Cargo (RNA with MS2 stem loop(s)) was packaged inside the particle either by producer cells expressing cargo and gag/PH and MS2 fused to DmrA or DmrC in the presence of A/C heterodimerizer, or particles being loaded by various particle loading methods described herein, such as electroporation.
[0162] FIG. 64: Depiction of exemplary heVLP and cargo configuration
[0163] This particle was created by producer cells expressing an envelope protein. Cargo (RNA with RBP stem loop(s)) was packaged inside the particle either by producer cells expressing cargo fused to an RBP and gag fused to another RBP or particles being loaded by various particle loading methods described herein, such as electroporation.
[0164] FIG. 65: Depiction of exemplary heVLP and cargo configuration
[0165] This particle was created by producer cells expressing an envelope protein. Cargo (RNA with RBP stem loop(s)) was packaged inside the particle either by producer cells expressing cargo fused to an RBP and PH fused to another RBP or particles being loaded by various particle loading methods described herein, such as electroporation.
[0166] FIG. 66: Depiction of exemplary heVLP and cargo configuration
[0167] This particle was created by producer cells expressing an envelope protein. Cargo (RNA with RBP stem loop(s)) was packaged inside the particle either by producer cells expressing cargo fused to an RBP and gag/PH fused to another RBP or particles being loaded by various particle loading methods described herein, such as electroporation.
[0168] FIG. 67: Depiction of exemplary heVLP and cargo configuration
[0169] This particle was created by producer cells expressing an envelope protein. Cargo (RNA with RBP stem loop(s)) was packaged inside the particle either by producer cells expressing cargo fused to an RBP and gag and another RBP fused to DmrA or DmrC in the presence of A/C Heterodimerizer molecule, or particles being loaded by various particle loading methods described herein, such as electroporation.
[0170] FIG. 68: Depiction of exemplary heVLP and cargo configuration
[0171] This particle was created by producer cells expressing an envelope protein. Cargo (RNA with RBP stem loop(s)) was packaged inside the particle either by producer cells expressing cargo fused to an RBP and PH and another RBP fused to DmrA or DmrC in the presence of A/C Heterodimerizer molecule, or particles being loaded by various particle loading methods described herein, such as electroporation.
[0172] FIG. 69: Depiction of exemplary heVLP and cargo configuration
[0173] This particle was created by producer cells expressing an envelope protein. Cargo (RNA with RBP stem loop(s)) was packaged inside the particle either by producer cells expressing cargo fused to an RBP and gag/PH and another RBP fused to DmrA or DmrC in the presence of A/C Heterodimerizer molecule, or particles being loaded by various particle loading methods described herein, such as electroporation.
DETAILED DESCRIPTION
[0174] Therapeutic proteins and nucleic acids hold great promise, but for many of these large biomolecules delivery into cells is a hurdle to clinical development. Genome editing reagents such as zinc finger nucleases (ZFNs) or RNA-guided, enzymatically active/inactive DNA binding proteins such as Cas9 have undergone rapid advancements in terms of specificity and the types of edits that can be executed, but the hurdle of safe in vivo delivery still precludes efficacious gene editing therapies. The following details the characteristics of the heVLP that make it a novel and optimal platform for the delivery of genome editing reagents, and contrasts heVLPs with canonical delivery modalities.
[0175] Retroviral particles, such as lentivirus, have been developed to deliver RNA that is reverse transcribed to DNA that may or may not be integrated into genomic DNA. VLPs have been developed that mimic virus particles in their ability to self-assemble, but are not infectious as they lack some of the core viral genes. Both lentiviral and VLP vectors are typically produced by transiently transfecting a producer cell line with plasmids that encode all components necessary to produce lentiviral particles or VLP. One major flaw that we have discovered regarding lentiviral particles and VSVG-based VLPs that are produced by this conventional transient transfection method is that, in addition to their conventional cargo, these particles package and deliver plasmid DNA that was used in the initial transient transfection. This unintended plasmid DNA delivery can be immunogenic and cause undesirable effects, such as plasmid DNA being integrated into genomic DNA. It is important to specify the type of biomolecules and/or chemicals that are to be delivered within particles, and heVLPs have been designed to possess this germane capability.
[0176] The heVLPs described herein can deliver DNA only, DNA+RNA+protein, or RNA+protein. Importantly, heVLPs are the first VLP delivery modality that leverages select components from human endogenous retroviruses (HERVs) to create particles for customizable cargo delivery into eukaryotic cells. heVLPs are capable of controlling the form of the cargo (DNA, protein, and/or RNA). All other previously described VLPs and viral particles package and deliver unwanted plasmid DNA (or other types of DNA-based gene expression constructs) introduced into particle producer cells via transient transfection in addition to the intended protein and/or RNA cargo(s).
[0177] Another non-obvious aspect of heVLPs is the ENV protein on the surface of the heVLP. The ENV protein is responsible for the ability of heVLPs to efficiently deliver cargo into cells. The majority of retroviral ENV proteins require post-translational modifications in the form of proteolytic cleavage of the intracellular domain (ICD) of the ENV protein in order to activate the fusogenicity of the ENV protein; this is essential for infectivity..sup.1 The envelope proteins described in Table 1 are all derived from HERVs that are expressed to varying levels in healthy human tissues (or HERV ENV consensus sequences). Some of these sequences possess ICD truncations that have been shown to enhance fusogenicity, but most do not require truncation.
[0178] heVLPs do not require exogenous, virally-derived GAG for particle formation because heVLPs utilize human-endogenous GAG proteins from HERVs (or HERV GAG consensus sequences)..sup.1 These HERV GAG proteins enable heVLP formation and are expressed to varying levels in healthy human tissues. Importantly, heVLPs are different from previously described viral particles, VLPs, and extracellular vesicles because heVLPs are composed of a novel combination of HERV ENV and GAG components, and heVLPs lack components from exogenous viruses..sup.2,3 Because of the above mentioned design optimizations, heVLPs are particularly suited for delivery of DNA, RNA, protein, or combinations of biomolecules and/or chemicals, such as DNA-encoded or RNP-based genome editing reagents.
[0179] Genome editing reagents, especially CRISPR-CAS, zinc finger, and TAL-nuclease-based reagents have the potential to become in vivo therapeutics for the treatment of genetic diseases, but techniques for delivering genome editing reagents into cells are severely limiting or unsafe for patients. Conventional therapeutic monoclonal antibody delivery is successful at utilizing direct injection for proteins. Unfortunately, strategies for direct injection of gene editing proteins, such as Cas9, are hampered by immunogenicity, degradation, ineffective cell specificity, and inability to cross the plasma membrane or escape endosomes/lysosomes..sup.4-10 More broad applications of protein therapy and gene editing could be achieved by delivering therapeutic protein cargo to the inside of cells. Cas9, for example, cannot efficiently cross the phospholipid bilayer to enter into cells, and has been shown to have innate and adaptive immunogenic potential..sup.4-8 Therefore, it is not practical or favorable to deliver Cas9 by direct injection or as an external/internal conjugate to lipid, protein or metal-based nanoparticles that have cytotoxic and immunogenic properties and often yield low levels of desired gene modifications..sup.9-20
[0180] Nanoparticles that encapsulate cargo are another delivery strategy that can be used to deliver DNA, protein, RNA and RNPs into cells.sup.9-18 Nanoparticles can be engineered for cell specificity and can trigger endocytosis and subsequent endosome lysis. However, nanoparticles can have varying levels of immunogenicity due to an artificially-derived vehicle shell..sup.9-20 Many nanoparticles rely on strong opposing charge distributions to maintain particle structural integrity, and the electrostatics can make it toxic and unfit for many in vivo therapeutic scenarios..sup.9 Nanoparticles that deliver RNA have had successes in recent clinical trials, but most have only been used to deliver siRNA or shRNA. Toxicity from such nanoparticles is still a major concern..sup.9 Nanoparticles that deliver mRNA coding for genome editing RNPs have also been a recent success, but these create a higher number of off-target effects compared to protein delivery and RNA stability is lower than that of protein..sup.17 Nanoparticles that deliver genome editing RNPs and DNA have been a significant breakthrough because they can leverage both homology directed repair (HDR) and non-homologous end joining (NHEJ), but exhibit prohibitively low gene modification frequencies in vitro and in vivo, and therefore currently have limited applications in vivo as a gene editing therapeutic..sup.15
[0181] Currently, the clinical standard vehicles for delivering genome editing therapeutics are adeno-associated virus (AAV). Although AAV vectors are a promising delivery modality that can successfully deliver DNA into eukaryotic cells, AAV cannot efficiently package and deliver DNA constructs larger than 4.5 kb and this precludes delivery of many CRISPR-based gene editing reagents that require larger DNA expression constructs. CRISPR-based gene editing reagents can be split into multiple different AAV particles, but this strategy drastically reduces delivery and editing efficiency. Depending on the dose required, AAV and adenoviral vectors can have varying levels of immunogenicity. In addition, inverted-terminal repeats (ITRs) in the AAV DNA construct can promote the formation of spontaneous episomes leading to prolonged expression of genome editing reagents and increased off-target effects. ITRs can also promote the undesired integration of AAV DNA into genomic DNA..sup.21-24
[0182] Recently, VLPs have been utilized to deliver mRNA and protein cargo into the cytosol of cells..sup.2,3,25-30 VLPs have emerged as a substitute delivery modality for retroviral particles. VLPs can be designed to lack the ability to integrate retroviral DNA, and to package and deliver protein/RNP/DNA. However, most VLPs, including recently conceived VLPs that deliver genome editing reagents known to date, utilize HIV or other virally-derived gag-pol protein fusions and viral proteases to generate retroviral-like particles..sup.25-27,29,30 Secondly, some VLPs containing RGNs also must package and express guide RNAs from a lentiviral DNA transcript..sup.27 Thirdly, some VLPs require a viral protease in order to form functional particles and release genome editing cargo..sup.25-27,29 Since this viral protease recognizes and cleaves at multiple amino acid motifs, it can cause damage to the protein cargo which could be hazardous for therapeutic applications. Fourthly, most published VLP modalities that deliver genome editing proteins to date exhibit low in vitro and in vivo gene modification efficiencies due to low packaging and transduction efficiency..sup.25-27 Fifthly, the complex viral genomes utilized for these VLP components possess multiple reading frames and employ RNA splicing that could result in spurious fusion protein products being delivered..sup.25-27,29,30 Sixthly, the presence of reverse transcriptase, integrase, capsid and a virally-derived envelope protein in these VLPs is not ideal for most therapeutic applications because of immunogenicity and off target editing concerns. Lastly, most retroviral particles, such as lentiviral particles, are pseudotyped with VSVG and nearly all described VLPs that deliver genome editing reagents hitherto possess and rely upon VSVG..sup.2,3,25-30 We have discovered that VSVG-based particles that are formed by transiently transfecting producer cells package and deliver DNA that was transfected. The current versions of VSVG-based VLPs cannot prevent this inadvertent delivery of DNA and this impedes the use of VLPs in scenarios that necessitate minimal immunogenicity and off target effects.
[0183] Extracellular vesicles are another delivery modality that can package and deliver cargo within exosomes and ectosomes..sup.31,32 Similar to VLPs, extracellular vesicles are comprised of a phospholipid bilayer from a mammalian cell. Unlike VLPs, extracellular vesicles lack viral components and therefore have limited immunogenicity. Whereas VLPs have a great ability to enter cells due to external fusogenic glycoproteins (VSVG) extracellular vesicles mainly rely on cellular uptake via micropinocytosis and this limits the delivery efficiency of extracellular vesicles.
[0184] heVLPs try to leverage the delivery benefits of extracellular vesicles and VLPs. heVLPs are the first VLP modality to eliminate all the potentially harmful exogenous, virally-derived components. heVLP components are known to be involved in extracellular vesicle biogenesis, they are known to possess local immunosuppressive properties, and their expression in healthy human tissues minimizes the chance of eliciting an immune response because of central tolerance..sup.1 heVLPs are a safer and more effective alternative than previously described VLPs, extracellular vesicles, AAVs and nanoparticles-especially for delivery of genome editing reagents-because heVLPs are comprised of all human-derived components, heVLPs have the ability to deliver DNA+RNP, or RNP alone while other previously described VLPs cannot prevent transient transfection DNA from being unintentionally packaged and delivered, heVLPs can deliver specialty DNA molecules while previously described VLPs, nanoparticles and AAVs cannot or do not, and heVLPs can be produced with cells that have been derived from patients (autologous heVLPs) and other FDA-approved cell lines (allogenic heVLPs) to further reduce the risks of adverse immune reactions. Here, we describe methods and compositions for producing, purifying, and administering heVLPs for in vitro and in vivo applications of genome editing, epigenome modulation, transcriptome editing and proteome modulation. The desired editing outcome depends on the therapeutic context and will require different gene editing reagents. Streptococcus pyogenes Cas9 (spCas9) and Acidaminococcus sp. Cas12a (functionalize) are two of the most popular RNA-guided enzymes for editing that leverages NHEJ for introducing stop codons or deletions, or HDR for causing insertions..sup.34-36 Cas9-deaminase fusions, also known as base editors, are the current standard for precise editing of a single nucleotide without double stranded DNA cleavage..sup.37,38 Importantly, this invention provides a novel way of packaging and delivering reagents for applications of genome editing, epigenome modulation, transcriptome editing and proteome modulation. Importantly, this invention is also the first to address the phenomenon of inadvertent DNA delivery in VLPs and the first to control for the type of biomolecule to be delivered (DNA, RNA, and/or protein) thereby increasing the types of therapeutic in vivo genome modifications that are possible and minimizing deleterious off target effects.
[0185] Section 1: heVLP-Mediated Delivery of Cargo Including DNAs, Proteins, Compounds, and RNAs
[0186] Conventional VLPs that have been engineered to encapsulate and deliver protein-based cargo commonly fuse cargo to the INT or GAG polyprotein..sup.25-27,29,30,39,40 After transient transfection of production plasmid DNA constructs, these protein fusions are translated in the cytosol of conventional VLP production cell lines, the gag matrix is acetylated and recruited to the cell membrane, and the gag fusions are encapsulated (transient transfection DNA is also unintentionally encapsulated) within VLPs as VLPs bud off of the membrane into extracellular space.
[0187] In contrast, the heVLPs described herein can package protein-based cargo by integrating all production DNA into the genomic DNA of production cell lines. Once cell lines are created, protein delivery heVLPs can be produced in a constitutive or inducible fashion. Proteins are packaged into heVLP by fusing select human-endogenous GAG proteins or other plasma membrane recruitment domains to protein-based cargo (e.g., as shown in Table 6). Human-endogenous GAG proteins and human pleckstrin homology (PH) domains localize to biological membranes. PH domains interact with phosphatidylinositol lipids and proteins within biological membranes, such as PIP2, PIPS, .beta..gamma.-subunits of GPCRs, and PKC..sup.41,42 However, in addition to localizing to phospholipid bilayers, human-endogenous GAG proteins drive budding and particle formation..sup.42 This dual functionality of human-endogenous GAG enables packaging of cargo and budding/formation of particles. One such human-endogenous GAG protein used for this purpose is the human Arc protein can be fused to protein-based cargo to recruit cargo to the cytosolic side of the phospholipid bilayer..sup.43 These human-endogenous GAG phospholipid bilayer recruitment domains can be fused to the N-terminus or C-terminus of protein-based cargo via polypeptide linkers of variable length regardless of the location or locations of one or more nuclear localization sequence(s) (NLS) within the cargo. Preferably, the linker between protein-based cargo and the human-endogenous GAG phospholipid bilayer recruitment domain is a polypeptide linker 5-20, e.g., 8-12, e.g., 10, amino acids in length primarily composed of glycines and serines. The human-endogenous GAG or other phospholipid bilayer recruitment domain localizes the cargo to the phospholipid bilayer and this protein cargo is packaged within heVLPs that bud off from the producer cell into extracellular space (FIG. 1). In this application, the use of these human-endogenous GAG and other phospholipid bilayer recruitment domains is novel and unique in that these human-endogenous GAG and other proteins can facilitate for localization of cargo to the cytosolic face of the plasma membrane within the heVLP production cells, and they also allow for cargo to localize to the nucleus of heVLP-transduced cells without the utilization of exogenous retroviral GAG or chemical and/or light-based dimerization systems (FIG. 2). The heVLP delivery of Cas9, for example, is significantly more efficient with a fusion to a human-endogenous GAG protein compared to a fusion to a PH plasma membrane recruitment domain or no fusion at all (FIG. 3).
[0188] heVLPs can also package and deliver a combination of DNA and RNA if heVLPs are produced via transient transfection of a production cell line. DNA that is transfected into cells will possess size-dependent mobility such that a fraction of the transfected DNA will remain in the cytosol while another fraction of the transfected DNA will localize to the nucleus..sup.44-46 One fraction of the transfected DNA in the nucleus will expressed components needed to create heVLPs and the other fraction in the cytosol/near the plasma membrane will be encapsulated and delivered in heVLPs (FIG. 4).
[0189] heVLP "Cargo" as used herein can refer to a one or more of chemicals, e.g., small molecule compounds, combination of DNA, RNA, and protein, a combination of RNA and protein, a combination of DNA and protein, or protein, e.g., for therapeutic or diagnostic use, or for the applications of genome editing, epigenome modulation, and/or transcriptome modulation. In addition, endogenous RNA and protein from the producer cells get packaged and/or incorporated into heVLPs. In order to simplify these distinctions, a combination of exogenous DNA, exogenous RNA, and protein (exogenous and/or endogenous protein) will be referred to as type 1 cargo (T1heVLPs), exogenous RNA and protein (exogenous and/or endogenous protein) will be referred to as type 2 cargo (T2heVLPs), a combination of exogenous DNA and proteins (exogenous and/or endogenous protein) will be referred to as type 3 cargo (T3heVLPs), proteins (exogenous and/or endogenous protein) will be referred to as type 4 cargo (T4heVLPs). Therefore, T1 contains DNA, RNA, +/-exogenous protein, T2 contains RNA+/-exogenous protein, T3 contains DNA+/-exogenous protein, and T4 is a particle with or without exogenous protein cargo. Hence, T4 without exogenous protein is considered an "empty particle" because there is no "exogenous cargo." "Exogenous cargo" is cargo not endogenous to the producer cells that can be packaged and/or incorporated into heVLPs. In addition, T1-T4heVLPs can package exogenous chemical molecules in addition to the types of cargoes present in T1-T4heVLPs. RNA in this context, for example, could be single guide RNA (sgRNA), Clustered Regularly Interspaced Palindromic Repeat (CRISPR) RNA (crRNA), and/or mRNA coding for cargo.
[0190] As used herein, "small molecules" refers to small organic or inorganic molecules of molecular weight below about 3,000 Daltons. In general, small molecules useful for the invention have a molecular weight of less than 3,000 Daltons (Da). The small molecules can be, e.g., from at least about 100 Da to about 3,000 Da (e.g., between about 100 to about 3,000 Da, about 100 to about 2500 Da, about 100 to about 2,000 Da, about 100 to about 1,750 Da, about 100 to about 1,500 Da, about 100 to about 1,250 Da, about 100 to about 1,000 Da, about 100 to about 750 Da, about 100 to about 500 Da, about 200 to about 1500, about 500 to about 1000, about 300 to about 1000 Da, or about 100 to about 250 Da).
[0191] The cargo is limited by the diameter of the particles, e.g., which in some embodiments range from 150 nm to 500 nm.
[0192] Cargo developed for applications of genome editing also includes nucleases and base editors. Nucleases include FokI and AcuI ZFNs and Transcription activator-like effector nucleases (TALENs) and CRISPR based nucleases or a functional derivative thereof (e.g., as shown in Table 2) (ZFNs are described, for example, in United States Patent Publications 20030232410; 20050208489; 20050026157; 20050064474; 20060188987; 20060063231; and International Publication WO 07/014275) (TALENs are described, for example, in United States Patent Publication U.S. Pat. No. 9,393,257B2; and International Publication WO2014134412A1) (CRISPR based nucleases are described, for example, in United States Patent Publications U.S. Pat. No. 8,697,359B1; US20180208976A1; and International Publications WO2014093661A2; WO2017184786A8)..sup.34-36 Base editors that are described by this work include any CRISPR based nuclease orthologs (wt, nickase, or catalytically inactive (CI)), e.g., as shown in Table 2, fused at the N-terminus to a deaminase or a functional derivative thereof (e.g., as shown in Table 3) with or without a fusion at the C-terminus to one or multiple uracil glycosylase inhibitors (UGIs) using polypeptide linkers of variable length (Base editors are described, for example, in United States Patent Publications US20150166982A1; US20180312825A1; U.S. Ser. No. 10/113,163B2; and International Publications WO2015089406A1; WO2018218188A2; WO2017070632A2; WO2018027078A8; WO2018165629A1)..sup.37,38 In addition, prime editors are also compatible with heVLP delivery modalities (Prime editors are described, for example, in Anzalone et al., Nature. 2019 December; 576(7785):149-157).
[0193] sgRNAs complex with genome editing reagents during the packaging process and are co-delivered within heVLPs. To date, this concept has been validated in vitro by experiments that demonstrate the T2heVLP delivery of RGN RNP for the purposes of site specific editing of an endogenous site (FIG. 3). For example, T2heVLPs have been used to deliver Cas9 RNP to HEK 293T cells for the purposes of editing endogenous VEGF site #3 (FIG. 3).
[0194] Cargo designed for the purposes of epigenome modulation includes the CI CRISPR based nucleases, zinc fingers (ZFs) and TALEs fused to an epigenome modulator or combination of epigenome modulators or a functional derivative thereof connected together by one or more variable length polypeptide linkers (Tables 2 & 4). T1-T4 cargo designed for the purposes of transcriptome editing includes CRISPR based nucleases or any functional derivatives thereof in Table 5 or CI CRISPR based nucleases or any functional derivatives thereof in Table 5 fused to deaminases in Table 3 by one or more variable length polypeptide linkers.
[0195] The cargo can also include any therapeutically or diagnostically useful protein, DNA, RNP, or combination of DNA, protein and/or RNP. See, e.g., WO2014005219; U.S. Ser. No. 10/137,206; US20180339166; U.S. Pat. No. 5,892,020A; EP2134841B1; WO2007020965A1. For example, cargo encoding or composed of nuclease or base editor proteins or RNPs or derivatives thereof can be delivered to retinal cells for the purposes of correcting a splice site defect responsible for Leber Congenital Amaurosis type 10. In the mammalian inner ear, heVLP delivery of base editing reagents or HDR promoting cargo to sensory cells such as cochlear supporting cells and hair cells for the purposes of editing .beta.-catenin (.beta.-catenin Ser 33 edited to Tyr, Pro, or Cys) in order to better stabilize .beta.-catenin could help reverse hearing loss.
[0196] In another application, heVLP delivery of RNA editing reagents or proteome perturbing reagents could cause a transitory reduction in cellular levels of one or more specific proteins of interest (potentially at a systemic level, in a specific organ or a specific subset of cells, such as a tumor), and this could create a therapeutically actionable window when secondary drug(s) could be administered (this secondary drug is more effective in the absence of the protein of interest or in the presence of lower levels of the protein of interest). For example, heVLP delivery of RNA editing reagents or proteome perturbing reagents could trigger targeted degradation of MAPK and PI3K/AKT proteins and related mRNAs in vemurafenib/dabrafenib-resistant BRAF-driven tumor cells, and this could open a window for the administration of vemurafenib/dabrafenib because BRAF inhibitor resistance is temporarily abolished (resistance mechanisms based in the MAPK/PI3K/AKT pathways are temporarily downregulated by heVLP cargo). This example is especially pertinent when combined with heVLPs that are antigen inducible and therefore specific for tumor cells.
[0197] In another application, heVLPs could deliver Yamanaka factors Oct3/4, Sox2, Klf4, and c-Myc to human or mouse fibroblasts in order to generate induced pluripotent stem cells.
[0198] In another application, heVLPs could deliver dominant-negative forms of proteins in order to elicit a therapeutic effect.
[0199] heVLPs that are antigen-specific could be targeted to cancer cells in order to deliver proapoptotic proteins BIM, BID, PUMA, NOXA, BAD, BIK, BAX, BAK and/or HRK in order to trigger apoptosis of cancer cells.
[0200] 90% of pancreatic cancer patients present with unresectable disease. Around 30% of patients with unresectable pancreatic tumors will die from local disease progression, so it is desirable to treat locally advanced pancreatic tumors with ablative radiation, but the intestinal tract cannot tolerate high doses of radiation needed to cause tumor ablation. Selective radioprotection of the intestinal tract enables ablative radiation therapy of pancreatic tumors while minimizing damage done to the surrounding gastrointestinal tract. To this end, heVLPs could be loaded with dCas9 fused to the transcriptional repressor KRAB and guide RNA targeting EGLN. EGLN inhibition has been shown to significantly reduce gastrointestinal toxicity from ablative radiation treatments because it causes selective radioprotection of the gastrointestinal tract but not the pancreatic tumor..sup.47
[0201] Unbound steroid receptors reside in the cytosol. After binding to ligands, these receptors will translocate to the nucleus and initiate transcription of response genes. heVLPs could deliver single chain variable fragment (scFv) antibodies to the cytosol of cells that bind to and disrupt cytosolic steroid receptors. For example, the scFv could bind to the glucocorticoid receptor and prevent it from binding dexamethasone, and this would prevent transcription of response genes, such as metallothionein IE which has been linked to tumorigenesis..sup.48
[0202] heVLPs can be indicated for treatments that involve targeted disruption of proteins. For example, heVLPs can be utilized for targeting and disrupting proteins in the cytosol of cells by delivering antibodies/scFvs to the cytosol of cells. Classically, delivery of antibodies through the plasma membrane to the cytosol of cells has been notoriously difficult and inefficient. This mode of protein inhibition is similar to how a targeted small molecule binds to and disrupts proteins in the cytosol and could be useful for the treatment of a diverse array of diseases..sup.49-51
[0203] In addition, the targeting of targeted small molecules is limited to proteins of a certain size that contain binding pockets which are relevant to catalytic function or protein-protein interactions. scFvs are not hampered by these limitations because scFvs can be generated that bind to many different moieties of a protein in order to disrupt catalysis and interactions with other proteins. For example, RAS oncoproteins are implicated across a multitude of cancer subtypes, and RAS is one of the most frequently observed oncogenes in cancer. For instance, the International Cancer Genome Consortium found KRAS to be mutated in 95% of their Pancreatic Adenocarcinoma samples. RAS isoforms are known to activate a variety of pathways that are dysregulated in human cancers, like the PI3K and MAPK pathways. Despite the aberrant roles RAS plays in cancer, no efficacious pharmacologic direct or indirect small molecule inhibitors of RAS have been developed and approved for clinical use. One strategy for targeting RAS could be heVLPs that can deliver specifically to cancer cells scFvs that bind to and disrupt the function of multiple RAS isoforms..sup.49-51
[0204] FIGS. 5-69 provide exemplary heVLP configurations and non-limiting examples of cargo molecules.
[0205] Section 2: heVLP Composition, Production, Purification and Applications
[0206] heVLPs are produced from producer cell lines that are either transiently transfected with at least one plasmid or stably expressing constructs that have been integrated into the producer cell line genomic DNA. In some embodiments, for T1 and T3heVLPs, if a single plasmid is used in the transfection, it should comprise sequences encoding one or more HERV-derived glycoproteins (e.g., as shown in Table 1), one or more HERV-derived GAG proteins, cargo (e.g., a therapeutic protein or a gene editing reagent such as a zinc finger, transcription activator-like effector (TALE), and/or CRISPR-based genome editing/modulating protein and/or RNP such as those found in Tables 2, 3, 4 & 5) with a fusion to a human-endogenous GAG or other plasma membrane recruitment domain (e.g., as shown in Table 6), and a guide RNA, if necessary. Preferably, two to three plasmids are used in the transfection. These two to three plasmids can include the following (any two or more can be combined in a single plasmid):
[0207] 1. A plasmid comprising sequences encoding a therapeutic protein or a genome editing reagent, with a fusion to a human-endogenous GAG or other plasma membrane recruitment domain.
[0208] 2. A plasmid comprising one or more HERV-derived glycoproteins (e.g., as listed in Table 1).
[0209] 3. A plasmid comprising one or more HERV-derived GAG proteins.
[0210] 4. If the genome editing reagent from plasmid 1 requires one or more guide RNAs, a plasmid comprising one or more guide RNAs apposite for the genome editing reagent in plasmid 1. If it is desired to deliver a type of DNA molecule other than plasmid(s), the above-mentioned transfection can be performed with double-stranded closed-end linear DNA, episome, mini circle, double-stranded oligonucleotide and/or other specialty DNA molecules. Alternatively, for T2 and T4heVLPs, the producer cell line can be made to stably express the constructs (1 through 3) described in the transfection above.
[0211] The plasmids, or other types of specialty DNA molecules described above, will also preferably include other elements to drive expression or translation of the encoded sequences, e.g., a promoter sequence; an enhancer sequence, e.g., 5' untranslated region (UTR) or a 3' UTR; a polyadenylation site; an insulator sequence; or another sequence that increases or controls expression (e.g., an inducible promoter element).
[0212] Preferably, appropriate producer cell lines are primary or stable human cell lines refractory to the effects of transfection reagents and fusogenic effects due glycoproteins. Examples of appropriate cell lines include Human Embryonic Kidney (HEK) 293 cells, HEK293 T/17 SF cells kidney-derived Phoenix-AMPHO cells, and placenta-derived BeWo cells. For example, such cells could be selected for their ability to grow as adherent cells, or suspension cells. In some embodiments, the producer cells can be cultured in classical DMEM under serum conditions, serum-free conditions, or exosome-free serum conditions. T1 and T3heVLPs can be produced from cells that have been derived from patients (autologous heVLPs) and other FDA-approved cell lines (allogenic heVLPs) as long as these cells can be transfected with DNA constructs that encode the aforementioned heVLP production components by various techniques known in the art.
[0213] In addition, if it is desirable, more than one genome editing reagent can be included in the transfection. The DNA constructs can be designed to overexpress proteins in the producer cell lines. The plasmid backbones, for example, used in the transfection can be familiar to those skilled in the art, such as the pCDNA3 backbone that employs the CMV promoter for RNA polymerase II transcripts or the U6 promoter for RNA polymerase III transcripts. Various techniques known in the art may be employed for introducing nucleic acid molecules into producer cells. Such techniques include chemical-facilitated transfection using compounds such as calcium phosphate, cationic lipids, cationic polymers, liposome-mediated transfection, such as cationic liposome like LIPOFECTAMINE (LIPOFECTAMINE 2000 or 3000 and TranslT-X2), polyethyleneimine, non-chemical methods such as electroporation, particle bombardment, or microinjection.
[0214] A human producer cell line that stably expresses the necessary heVLP components in a constitutive and/or inducible fashion can be used for production of T2 and T4heVLPs. T2 and T4heVLPs can be produced from cells that have been derived from patients (autologous heVLPs) and other FDA-approved cell lines (allogenic heVLPs) if these cells have been converted into stable cell lines that express the aforementioned heVLP components.
[0215] Also provided herein are the producer cells themselves.
[0216] In some embodiments, in order for efficient recruitment of cargo into heVLPs, the cargo comprises a covalent or non-covalent connection to a human-endogenous GAG or other plasma membrane recruitment domain, preferably as shown in Table 6. Covalent connections, for example, can include direct protein-protein fusions generated from a single reading frame, inteins that can form peptide bonds, other proteins that can form covalent connections at R-groups and/or RNA splicing..sup.52-54 Non-covalent connections, for example, can include DNA/DNA, DNA/RNA, and/or RNA/RNA hybrids (nucleic acids base pairing to other nucleic acids via hydrogen-bonding interactions), protein domains that dimerize or multimerize with or without the need for a chemical compound/molecule to induce the protein-protein binding (such as DmrA/DmrB/DmrC (Takara Bio), FKBP/FRB,.sup.55 dDZFs,.sup.56 and Leucine zippers.sup.57), single chain variable fragments,.sup.58 nanobodies,.sup.59 affibodies,.sup.60 proteins that bind to DNA and/or RNA, proteins with quaternary structural interactions, optogenetic protein domains that can dimerize or multimerize in the presence of certain light wavelengths,.sup.61 and/or naturally reconstituting split proteins..sup.62
[0217] In some embodiments, the cargo comprises a fusion to a dimerization domain or protein-protein binding domain that may or may not require a molecule to trigger dimerization or protein-protein binding.
[0218] In some embodiments, the producer cells are FDA-approved cells lines, allogenic cells, and/or autologous cells derived from a donor.
[0219] In some embodiments, the full or active peptide domains of human CD47 may be incorporated in the heVLP surface to reduce immunogenicity.
[0220] Examples of AAV proteins included here are AAV REP 52, REP 78, and VP1-3. The capsid site where proteins can be inserted is T138 starting from the VP1 amino acid counting..sup.63 Dimerization domains could be inserted at this point in the capsid, for instance.
[0221] Examples of dimerization domains included here that may or may not need a small molecule inducer are dDZF1,.sup.56 dDZF2,.sup.56 DmrA (Takara Bio), DmrB (Takara Bio), DmrC (Takara Bio), FKBP,.sup.55 FRB,.sup.55 GCN4 scFv,.sup.58 10.times./24.times. GCN4,.sup.58 GFP nanobody.sup.59 and GFP..sup.64
[0222] Examples of split inteins included here are Npu DnaE, Cfa, Vma, and Ssp DnaE..sup.52
[0223] Examples of other split proteins included here that make a covalent bond together are Spy Tag and Spy Catcher..sup.53
[0224] Examples of RNA binding proteins included here are MS2, Com, and PP7..sup.65 Examples of synthetic DNA-binding zinc fingers included here are ZF6/10, ZF8/7, ZF9, MK10, Zinc Finger 268, and Zinc Finger 268/NRE..sup.66,67
[0225] Examples of proteins that multimerize as a result of quaternary structure included here are E. coli ferritin, and the other chimeric forms of ferritin..sup.68,69
[0226] Examples of optogenetic "light-inducible proteins" included here are Cry2, CIBN, and Lov2-Ja..sup.61
[0227] Examples of peptides the enhance transduction included here are L17E,.sup.70 Vectofusin-1 (Miltenyi Biotec), KALA,.sup.71 and the various forms of nisin..sup.72
[0228] In another embodiment, T1-T4 heVLPs that are produced and isolated can be loaded with biomolecule or chemical molecule cargo by utilizing nucleofection, lipid, polymer, or CaCl.sub.2 transfection, sonication, freeze thaw, incubation at various temperatures, and/or heat shock of purified particles mixed with cargo. These techniques are adapted from techniques employed to load cargo into exosomes for therapeutic or research applications..sup.73-75 For example, 100 ug of heVLPs can be resuspended in 200-450 ul of 50 mM trehalose in PBS, mixed with cargo at a desired concentration, and electroporated (GenePulser II Electroporation System with capacitance extender, Bio-Rad, Hercules, Calif., USA) in a 0.4 cm cuvette at 0.200 kV and 125 uF.
Production of Cargo-Loaded heVLPs and Compositions
[0229] Preferably heVLPs are harvested from cell culture medium supernatant 36-48 hours post-transfection, or when heVLPs are at the maximum concentration in the medium of the producer cells (the producer cells are expelling particles into the media and at some point in time, the particle concentration in the media will be optimal for harvesting the particles). Supernatant can be purified by any known methods in the art, such as centrifugation, ultracentrifugation, precipitation, ultrafiltration, and/or chromatography. In some embodiments, the supernatant is first filtered, e.g., to remove particles larger than 1 .mu.m, e.g., through 0.45 pore size polyvinylidene fluoride hydrophilic membrane (Millipore Millex-HV) or 0.8 .mu.m pore size mixed cellulose esters hydrophilic membrane (Millipore Millex-AA). After filtration, the supernatant can be further purified and concentrated, e.g., using ultracentrifugation, e.g., at a speed of 80,000 to 100,000.times.g at a temperature between 1.degree. C. and 5.degree. C. for 1 to 2 hours, or at a speed of 8,000 to 15,000 g at a temperature between 1.degree. C. and 5.degree. C. for 10 to 16 hours. After this centrifugation step, the heVLPs are concentrated in the form of a centrifugate (pellet), which can be resuspended to a desired concentration, mixed with transduction-enhancing reagents, subjected to a buffer exchange, or used as is. In some embodiments, heVLP-containing supernatant can be filtered, precipitated, centrifuged and resuspended to a concentrated solution. For example, polyethylene glycol (PEG), e.g., PEG 8000, or antibody-bead conjugates that bind to heVLP surface proteins or membrane components can be used to precipitate particles. Purified particles are stable and can be stored at 4.degree. C. for up to a week or -80.degree. C. for years without losing appreciable activity.
[0230] Preferably, heVLPs are resuspended or undergo buffer exchange so that particles are suspended in an appropriate carrier. In some embodiments, buffer exchange can be performed by ultrafiltration (Sartorius Vivaspin 500 MWCO 100,000). An exemplary appropriate carrier for heVLPs to be used for in vitro applications would preferably be a cell culture medium that is suitable for the cells that are to be transduced by heVLPs. Transduction-enhancing reagents that can be mixed into the purified and concentrated heVLP solution for in vitro applications include reagents known by those familiar with the art (Miltenyl Biotec Vectofusin-1, Millipore Polybrene, Takara Retronectin, Sigma Protamine Sulfate, and the like). After heVLPs in an appropriate carrier are applied to the cells to be transduced, transduction efficiency can be further increased by centrifugation. Preferably, the plate containing heVLPs applied to cells can be centrifuged at a speed of 1,150 g at room temperature for 30 minutes. After centrifugation, cells are returned into the appropriate cell culture incubator (humidified incubator at 37.degree. C. with 5% CO.sub.2).
[0231] An appropriate carrier for heVLPs to be administered to a mammal, especially a human, would preferably be a pharmaceutically acceptable composition. A "pharmaceutically acceptable composition" refers to a non-toxic semisolid, liquid, or aerosolized filler, diluent, encapsulating material, colloidal suspension or formulation auxiliary of any type. Preferably, this composition is suitable for injection. These may be in particular isotonic, sterile, saline solutions (monosodium or disodium phosphate, sodium, potassium, calcium or magnesium chloride and similar solutions or mixtures of such salts), or dry, especially freeze-dried compositions which upon addition, depending on the case, of sterilized water or physiological saline, permit the constitution of injectable solutions. Another appropriate pharmaceutical form would be aerosolized particles for administration by intranasal inhalation or intratracheal intubation.
[0232] The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or suspensions. The solution or suspension may comprise additives which are compatible with heVLPs and do not prevent heVLP entry into target cells. In all cases, the form must be sterile and must be fluid to the extent that the form can be administered with a syringe. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi. An example of an appropriate solution is a buffer, such as phosphate buffered saline.
[0233] Methods of formulating suitable pharmaceutical compositions are known in the art, see, e.g., Remington: The Science and Practice of Pharmacy, 21st ed., 2005; and the books in the series Drugs and the Pharmaceutical Sciences: a Series of Textbooks and Monographs (Dekker, NY). For example, solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.
[0234] Pharmaceutical compositions suitable for injectable use can include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL.TM. (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that easy syringability exists. It should be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyetheylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent that delays absorption, for example, aluminum monostearate and gelatin.
[0235] Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle, which contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying, which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.
[0236] The compositions comprising cargo-loaded heVLPs can be included in a container, pack, or dispenser together with instructions for administration.
EXAMPLES
[0237] The invention is further described in the following examples, which do not limit the scope of the invention described in the claims.
[0238] Methods
[0239] heVLP particles were produced by HEK293T cells using polyethylenimine (PEI) based transfection of plasmids. PEI is Polyethylenimine 25 kD linear (Polysciences #23966-2). To make a stock `PEI MAX` solution, 1 g of PEI was added to 1 L endotoxin-free dH.sub.2O that was previously heated to -80.degree. C. and cooled to room temperature. This mixture was neutralized to pH 7.1 by addition of 10N NaOH and filter sterilized with 0.22 .mu.m polyethersulfone (PES). PEI MAX is stored at -20.degree. C.
[0240] HEK293T cells were split to reach a confluency of 70%-90% at time of transfection and are cultured in 10% FBS DMEM media. Cargo vectors, such as one encoding a CMV promoter driving expression of a hPLC.delta.1 PH fusion to codon optimized Cas9 were co-transfected with a U6 promoter-sgRNA encoding plasmid, a hERVK.sub.conGAG (hGAGK.sub.con) encoding plasmid, and a hENVW (Syncytin-1) encoding plasmid. Transfection reactions were assembled in reduced serum media (Opti-MEM; GIBCO #31985-070). For heVLP particle production on 10 cm plates, 5 .mu.g PH-Cas9 expressing plasmid, 5 .mu.g sgRNA-expression plasmid, 5 .mu.g hERVK.sub.conGAG expression plasmid, and 5 .mu.g Syncytin-1 expression plasmid were mixed in 1 mL Opti-MEM, followed by addition of 27.5 .mu.l PEI MAX. After 20-30 min incubation at room temperature, the transfection reactions were dispersed dropwise over the HEK293T cells.
[0241] heVLPs were harvested at 48-72 hours post-transfection. heVLP supernatants were filtered using 0.8 .mu.m pore size mixed cellulose esters membrane filters and transferred to polypropylene Beckman ultracentrifuge tubes that are used with the SW28 rotor (Beckman Coulter #326823). Each ultracentrifuge tube is filled with heVLP-containing supernatant from 3 10 cm plates to reach an approximate final volume of 35-37.5 ml. heVLP supernatant underwent ultracentrifugation at approximately 100,000.times.g, or 25,000 rpm, at 4.degree. C. for 2 hours. After ultracentrifugation, supernatants were decanted and heVLP pellets resuspended in DMEM 10% FBS media such that they are now approximately 1,000 times more concentrated than they were before ultracentrifugation. heVLPs were added dropwise to cells that were seeded in a 24-well plate 24 hours prior to transduction. Polybrene (5-10 .mu.g/mL in cell culture medium; Sigma-Aldrich #TR-1003-G) was supplemented to enhance transduction efficiency, if necessary. Vectofusin-1 (10 .mu.g/mL in cell culture medium, Miltenyi Biotec #130-111-163) was supplemented to enhance transduction efficiency, if necessary. Immediately following the addition of heVLPs, the 24-well plate was centrifuged at 1,150.times.g for 30 min at room temperature to enhance transduction efficiency, if necessary.
Example 1
[0242] HEK 293T cells were transduced with T1heVLPs containing PLC PH fused to spCas9, hGAGK.sub.con fused to spCas9, or hArc fused to spCas9 targeted to VEGF site #3. T1heVLPs were pseudotyped with either hENVW (left chart) or hENVFRD (right chart). Gene modification was measured by amplicon sequencing. Particle purification and concentration was performed by PVDF filtration and ultracentrifugation at 100,000.times.g for 2 hours. Results are shown in FIG. 3. Importantly, if HERV-derived GAG (hGAGK.sub.con) was not overexpressed by itself in producer cells, then efficient delivery was not achieved.
TABLE-US-00001 TABLE 1 Position in # HERV envelope Gene name Accession no. sequence entry (a) 1. hENVH1 envH/p62 AJ289709.1 6313-8067 (+) 2. hENVH2 envH/p60 AJ289710.2 5393-7084 (+) 3. hENVH3 envH/p59 AJ289711.1 5204-6871 (+) 4. hENVK1 envK1 AC074261.3 93508-95604 (+) 5. hENVK2 envK2/HML-2.HOM AC072054.10 30365-32464 (-) 6. hENVK3 envK3/C19 Y17833.1 5581-7680 (+) 7. hENVK4 envK4/K109 AF164615.1 6412-8508 (+) 8. hENVK5 envK5/K113 AY037928.1 6451-8550 (+) 9. hENVK6 envK6/K115 AY037929.1 6442-8541 (+) 10. hENVT envT AC078899.1 154738-156618 (+) 11. hENVW Syncytin-1 AC000064.1 35879-37495 (+) 12. hENVFRD Syncytin-2 AL136139.6 21355-22972 (-) 13. hENVR erv-3 AC073210.8 54963-56978 (-) 14. hENVR(b) envRb AC093488.1 78681-80225 (+) 15. hENVF(c)2 envFc2 AC016222.4 85216-86963 (+) 16. hENVF(c)1 envFc1 AL354685.2 46744-48717 (-) *17. hENVK.sub.con N/A N/A N/A (a) `+` and `-` refer to the orientation within the sequence entry *hENVK.sub.con is a consensus sequence derived from ten proviral ENV sequences. The ENV sequences used to derive this consensus ENV sequence are from the following HERVs: HERV-K113, HERV-K101, HERV-K102, HERV-K104, HERV-K107, HERV-K108, HERV-K109, HERV-K115, HERV- K11p22, and HERV-K12q13.
TABLE-US-00002 TABLE 2 Exemplary Potential Cas9 and Cas12a orthologs DNA-binding Cas ortholog Enzyme class Nickase mutation CI mutations SpCas9 Type II-A D10A D10A, H840A SaCas9 Type II-A D10A D10A, CjCas9 Type II-C D8A D8A, NmeCas9 Type II-C D16A D16A, H588A asCas12a Type II-C D908A, E993A lbCas12a Type II-C D832A, E925A Nickase mutation residues represents a position of the enzyme either known to be required for catalytic activity of the conserved RuvC nuclease domain or predicted to be required for this catalytic activity based on sequence alignment to CjCas9 where structural information is lacking (* indicates which proteins lack sufficient structural information). All positional information refers to the wild-type protein sequences acquired from uniprot.org.
TABLE-US-00003 TABLE 3 Exemplary Deaminase domains and their substrate sequence preferences. Deaminase Nucleotide sequence preference hAID 5'-WRC rAPOBEC1* 5'-TC .gtoreq. CC .gtoreq. AC > GC mAPOBEC3 5'-TYC hAPOBEC3A 5'-TCG hAPOBEC3B 5'-TCR > TCT hAPOBEC3C 5'-WYC hAPOBEC3F 5'-TTC hAPOBEC3G 5'-CCC hAPOBEC3H 5'-TTCA ~ TTCT ~ TTCG > ACCCA > TGCA ecTadA hAdar1 hAdar2 Nucleotide positions that are poorly specified or are permissive of two or more nucleotides are annotated according to IUPACcodes, where W = A or T, R = A or G, and Y = C or T.
TABLE-US-00004 TABLE 4 Exemplary Epigenetic modulator domains. Epigenetic modulator Epigenetic modulation VP16 transcriptional activation VP64 transcriptional activation P65 transcriptional activation RTA transcriptional activation KRAB transcriptional repression MeCP2 transcriptional repression Tet1 Methylation Dnmt3a Methylation
TABLE-US-00005 TABLE 5 Exemplary CRISPR based RNA-guided RNA binding enzymes RNA-binding Cas ortholog Enzyme class LshCas13a Type-VI LwaCas13a Type-VI PspCas13b Type-VI RfxCas13d Type-VI
TABLE-US-00006 TABLE 6 Plasma membrane recruitment domains described in this work. # Plasma membrane recruitment domain Substitution(s) 1. Pleckstrin homology domain of human phospholipase C.delta.1 (hPLC.delta.1) 2. Pleckstrin homology domain of human Akt1 3. Mutant Pleckstrin homology domain of E17K human Akt1 4. hArc *5. hGAGK.sub.con 6. Pleckstrin homology domain of human 3- phosphoinositide-dependent protein kinase 1 (hPDPK1) 7. Human CD9 8. Human CD47 9. Human CD63 10. Human CD81 *hGAGK.sub.con is a consensus sequence derived from ten proviral GAG sequences. The GAG sequences used to derive this consensus GAG sequence are from the following HERVs: HERV-K113, HERV-K101, HERV-K102, HERV-K104, HERV-K107, HERV-K108, HERV-K109, HERV-K115, HERV- K11p22, and HERV-K12q13.
TABLE-US-00007 Relevant Protein Sequences: Homo sapiens: Arc (SEQ ID NO: 1) MELDHRTSGGLHAYPGPRGGQVAKPNVILQIGKCRAEMLEHVRRTHRHLLAEVSKQVERELKGLHRSVGKLE SNLDGYVPTSDSQRWKKSIKACLCRCQETIANLERWVKREMHVVVREVFYRLERWADRLESTGGKYPVGSES ARHTVSVGVGGPESYCHEADGYDYTVSPYAITPPPAAGELPGQEPAEAQQYQPWVPGEDGQPSPGVDTQIFE DPREFLSHLEEYLRQVGGSEEYWLSQIQNHMNGPAKKVWVEFKQGSVKVWVEFKKEFLQYSEGTLSREAIQR ELDLPQKQGEPLDQFLWRKRDLYQTLYVDADEEEIIQYVVGTLQPKLKRFLRHPLPKTLEQLIQRGMEVQDD LEQAAEPAGPHLPVEDEAETLTPAPNSESVASDRTQPE >AJ289709.1 Human endogenous retrovirus HHERV-H/env62 HERV_H/ENV_62 hENVH1: (SEQ ID NO: 2 MIFAGKAPSNTSTLMKFYSLLLYSLLFSFPFLCHPLPLPSYLHHTINLTHSLLAASNPSLVNNCWLCISLSS SAYTAVPAVQTDWATSPISLHLRTSFNSPHLYPPEELIYFLDRSSKTSPDISHQQAAALLRTYLKNLSPYIN STPPIFGPLTTQTTIPVAAPLCISWQRPTGIPLGNLSPSRCSFTLHLRSPTTNINETIGAFQLHITDKPSIN TDKLKNISSNYCLGRHLPCISLHPWLSSPCSSDSPPRPSSCLLIPSPENNSERLLVDTRRFLIHHENRTFPS TQLPHQSPLQPLTAAALAGSLGVVVVQDTPFSTPSHLFTLHLQFCLAQGLFFLCGSSTYMCLPANVVTGTCT LVFLTPKIQFANGTEELPVPLMTPTQQKRVIPLIPLMVGLGLSASTVALGTGIAGISTSVMTFRSLSNDFSA SITDISQTLSVLQAQVDSLAAVVLQNRRGLDLLTAEKGGLCIFLNEECCFYLNQSGLVYDNIKKLKDRAQKL ANQASNYAEPPWALSNWMSWVLPIVSPLIPIFLLLLFGPCIFRLVSQFIQNRIQAITNHSIRQMFLLTSPQY HPLPQDLPSA >AJ289710.2 Human endogenous retrovirus HHERV-H/env60- HERV_H_ENV_60-ENVH2: (SEQ ID NO: 3) MIFAGRASSNTSTLMKFYSLLLYSLLFSFPILCHPLPLPSYLHHTINLTHSLLAVSNPSLAKNCWLCISLPS SAYPAVPALQTDWGTSPVSPHLRTSFNSPHLYPPEKLIYFLDRSSKTSPDISHQQAAALLCTYLKNLSPYIN STPPTFGPLTTQTTIPVAAPLCISRQRPTGIPLGNLSPSRCSFTLHLRSPTTHITETNGAFQLHITDKPSIN TDKLKNVSSNYCLGRHLSCISLHPWLFSPCSSDSPPRPSSCLLIPSPKNNSESLLVDAQRFLIYHENRTSPS TQLPHQSPLQPLTAAPLGGSLRVVVVQDTPFSTPSHLFTLHLQFCLVQSLFFLCGSSTYMCLPANWTGTCTL VFLTSKIQFANGTEELPVPLMTPTRQKRVIPLIPLMVGLGLSASTVALGTGIAGISTSVTTFRILSNDFSAS ITDISQTLSGLQAQVDSSAAVVLQNRQGLDLLTAEKGGLCIFLNEESYFYLNQSGLVYDNIKKLKDKAQNLA NQASNYAEPPWPLSNWMSWVLPILSPLIPIFLLLFFRPCIFHLVSQFIQNHIQAITDHSI >AJ289711.1 Human endogenous retrovirus HHERV-H/env59- HERV_H_ENV_59-hENVH3: (SEQ ID NO: 4) MILAGRAPSNTSTLMKFYSLLLYSLLFSFPFLYHPLPLPSYLHHTINLTHSLPAASNPSLANNCWLCISLSS SAYIAVPTLQTDRATSPVSLHLRTSFNSPHLYPPEELIYFLDRSSKTSPDISHQPAAALLHIYLKNLSPYIN STPPIFGPLTTQTTIPVAAPLCISRQRPTGIPLGNISPSRCSFTLHLQSPTTHVTETIGVFQLHIIDKPSIN TDKLKNVSSNYCLGRHLPYISLHPWLPSPCSSDSPPRPSSCLLTPSPQNNSERLLVDTQRFLIHHENRTSSS MQLAHQSPLQPLTAAALAGSLGVWVQDTPFSTPSHPFSLHLQFCLTQGLFFLCGSSTYMCLPANWTGTCTLV FLTPKIQFANGTKELPVPLMTLTPQKRVIPLIPLMVGLGLSASTIALSTGIAGISTSVTTFRSPSNDFSASI TDISQTLSVLQAQVDSLAAVVLQNRRGLGLSILLNEECCFYLNQSGLVYENIKKLKDRAQKLANQASNYAES PWALSNWMSWVLPILSPLIPIFLLLLFGPCIFHLVSQFIQNRIQAITNHSI >AC074261.3 Homo sapiens chromosome 12 clone RP11-55F19 envK1-ENVK1: (SEQ ID NO: 5) MHPSEMQRKAPPRRRRHRNRAPLTHKMNKMVTSEQMKLPSTKKAEPPTWAQLKKLTQLATKYLENTKVTQTP ESMLLAALMIVSMVVSLPMPAGAAAANYTNWAYVPFPPLIRAVTWMDNPIEVYVNDSVWVHGPIDDRCPAKP EEEGMMINISIGYHYPPICLGRAPGCLMPAVQNWLVEVPTVSPISRFTYNMVSGMSLRPRVNYLQDFSYQRS LKFRPKGKPCPKEIPKESKNTEVLVWEECVANSVVILQNNEFGTIIDWAPRGQFYHNCSGQTQSCPSAQVSP AVDSDLTESLDKHKHKKLQSFYPWEWGEKGISTPRPKIISPVSGPEHPELWRLTVASHHIRIWSGNQTLETR DRKPFYTVDLNSSLTVPLQSCVKPPYMLVVGNIVIKPDSQTITCENCRLLTCIDSTFNWQHRILLVRAREGV WIPVSMDRPWEASPSIHILTEVLKGVLNRSKRFIFTLIAVIMGLIAVTAMAAVAGVALHSFVQSVNFVNDWQ KNSTRLWNSQSSIDQKLANQINDLRQTVIWMGDRLMSLEHRFQLQCDWNTSDFCITPQIYNESEHHWDMVRR HLQGREDNLTLDISKLKEQIFEASKAHLNLVPGTEAIAGVADGLANLNPVTWVKTIGSTTIINLILILVCLF CLLLVCRFTQQLRRDSYHRERAMMTMVVLSKRKGGNVGKSKRDQIVTVSV >AC072054.10 Homo sapiens BAC clone RP11-33P21-ENVK2: (SEQ ID NO: 6) MNPSEMQRKAPPRRRRHRNRAPLTHKMNKMVTSEEQMKLPSTKKAEPPTWAQLKKLTQLATKYLENTKVTQT PESMLLAALMIVSMVVSLPMPAGAAAANYTYWAYVPFPPLIRAVTWMDNPTEVYVNDSVWVPGPIDDRCPAK PEEEGMMINISIGYHYPPICLGRAPGCLMPAVQNWLVEVPTVSPICRFTYHMVSGMSLRPRVNYLQDFSYQR SLKFRPKGKPCPKEIPKESKNTEVLVWEECVANSAVILQNNEFGTIIDWAPRGQFYHNCSGQTQSCPSAQVS PAVDSDLTESLDKHKHKKLQSFYPWEWGEKGISTPRPKIVSPVSGPEHPELWRLTVASHHIRIWSGNQTLET RDRKPFYTIDLNSSLTVPLQSCVKPPYMLVVGNIVIKPDSQTITCENCRLLTCIDSTFNWQHRILLVRAREG VWIPVSMDRPWEASPSVHILTEVLKGVLNRSKRFIFTLIAVIMGLIAVTATAAVAGVALHSSVQSVNFVNDW QKNSTRLWNSQSSIDQKLANQINDLRQTVIWMGDRLMSLEHRFQLQCDWNTSDFCITPQIYNESEHHWDMVR RHLQGREDNLTLDISKLKEQIFEASKAHLNLVPGTEAIAGVADGLANLNPVTWVKTIGSTTIINLILILVC LFCLLLVCRCTQQLRRDSDHRERAMMTMAVLSKRKGGNVGKSKRDQIVTVSV >Y17833.1 Human endogenous retrovirus K (HERV-K) envK3-ENVK3: (SEQ ID NO: 7) MNPSEMQRKAPPRRRRHRNRAPLTHKMNKMVTSEEQMKLPSTKKAEPPTWAQLKKLTQLATKYLENTKVTQT PESMLLAALMIVSMVVSLPMPAGAAAANYTYWAYVPFPPLIRAVTWMDNPIEVYVNDSVWVPGPTDDHCPAK PEEEGMMINISIGYRYPPICLGRAPGCLMPAVQNWLVEVPTVSPISRFTYHMVSGMSLRPRVNYLQDFSYQR SFKFRPKGKPCPKEIPKESKNTEVLVWEECVANSAVILQNNEFGTIIDWAPRGQFYHNCSGQTQSCPSAQVS PAVDSDLTESLDKHKHKKLQSFYPWEWGEKGISTPRPKIISPVSGPEHPELWRLTVASHHIRIWSGNQTLET RDRKPFYTVDLNSSVTVPLQSCIKPPYMLVVGNIVIKPDSQTITCENCRLLTCIDSTFNWQHRILLVRAREG VWIPVSMDRPWETSPSIHTLTEVLKGVLNRSKRFIFTLIAVIMGLIAVTATAAVAGVALHSSVQSVNFVNDW QKNSTRLWNSQSSIDQKLANQINDLRQTVIWMGDRLMSLEHRFQLQCDWNTSDFSITPQIYNESEHHWDMVR RHLQGREDNLTLDISKLKEQIFEASKAHLNLVPGTEAIAGVADGLANLNPVTWVKTIGSTTIINLILILVCL FCLLLVCRCTQQLRRDSDHRERAMMTMAVLSKRKGGNVGKSKRDQIVTVSV >AF164615.1 Homo sapiens endogenous retrovirus HERV-K109 envK4-ENVK4: (SEQ ID NO: 8) MNPSEMQRKAPPRRRRHRNRAPLTHKMNKMVTSEEQMKLPSTKKAEPPTWAQLKKLTQLATKYLENTKVTQT PESMLLAALMIVSMVVSLPMPAGAAAANYTNWAYVPFPPLIRAVTWMDNPIEVYVNDSVWVPGPIDDRCPAK PEEEGMMINISIGYRYPICLGRAPGCLMPAVQNWLVEVPIVSPICRFTYHMVSGMSLRPRVNYLQDFSYQRS LKFRPKGKPCPKEIPKESKNTEVLVWEECVANSAVILQNNEFGTIIDVVTPQGQFYHNCSGQTQSCPSAQVS PAVDSDLTESLDKHKHKKLQSFYPWEWGEKGISTPRPKIISPVSGPEHPELWRLTVASHHIRIWSGNQTLET RDRKPFYTVDLNSSLTLPLQSCVKPPYMLVVGNIVIKPDSQTITCENCRLLTCIDSTFNWQHRILLVRAREG VWIPVSMDRPWEASPSIHILTEVLKGVLNRSKRFIFTLIAVIMGLIAVTATAAVAGVALHSSVQSVNFVNDG QKNSTRLWNSQSSIDQKLANQINDLRQTVIWMGDRLMSLEHRFQLQCDWNTSDFCITPQIYNESEHHWDMVR RHLQGREDNLTLDISKLKEQIFEASKAHLNLVPGTEAIAGVADGLANLNPVTWVKTIGSTTIINLILILVCL FCLLLVCRCTQQLRRDSDHRERAMMTMAVLSKRKGGNVGKSKRDQIVTVSV >AY037928.1 Human endogenous retrovirus K113 envK5-ENVK5: (SEQ ID NO: 9) MNPSEMQRKAPPRRRRHRNRAPLTHKMNKMVTSEEQMKLPSTKKAEPPTWAQLKKLTQLATKYLENTKVTQT PESMLLAALMIVSMVVSLPMPAGAAAANYTYWAYVPFPPLIRAVTWMDNPIEIYVNDSVWVPGPTDDCCPAK PEEEGMMINISIGYRYPPICLGRAPGCLMPAVQNWLVEVPTVSPISRFTYHMVSGMSLRPRVNYLQDFSYQR SLKFRPKGKPCPKEIPKESKNTEVLVWEECVANSAVILQNNEFGTLIDWAPRGQFYHNCSGQTQSCPSAQVS PAVDSDLTESLDKHKHKKLQSFYPWEWGEKGISTARPKIISPVSGPEHPELWRLTVASHHIRIWSGNQTLET RDRKPFYTIDLNSSLTVPLQSCVKPPYMLVVGNIVIKPDSQTITCENCRLLTCIDSTFNWQHRILLVRAREG VWIPVSMDRPWEASPSVHILTEVLKGVLNRSKRFIFTLIAVIMGLIAVTATAAVAGVALHSSVQSVNFVNDW QNNSTRLWNSQSSIDQKLANQINDLRQTVIWMGDRLMSLEHRFQLQCDWNTSDFCITPQIYNESEHHWDMVR CHLQGREDNLTLDISKLKEQIFEASKAHLNLVPGTEAIAGVADGLANLNTVTWVKTIGSTTIINLILILVCL FCLLLVYRCTQQLRRDSDHRERAMMTMVVLSKRKGGNVGKSKRDQIVTVSV >AY037929.1 Human endogenous retrovirus K115 envK6-ENVK6: (SEQ ID NO: 10) MNPSEMQRKAPPRRRRHRNRAPLTHKMNKMVTSEEQMKLPSTKKAEPPTWAQLKKLTQLATKYLENTKVTQT PESMLLAALMIVSMVVSLPMPAGAAVANYTNWAYVPFPPLIRAVTWMDNPIEVYVNDSVWVPGPIDDRCPAK PEEEGMMINISIGYRYPPICLGRAPGCLMPAVQNWLVEVPTVSPISRFTYHMVSGMSLRPRVNYLQDFSYQR SLKFRPKGKPCPKEIPKESKNTEVLVWEECVANSAVILQNNEFGTIIDWAPRGQFYHNCSGQTQSCPSAQVS PAVDSDLTESLDKHKHKKLQSFYPWEWGEKRISTPRPKIVSPVSGPEHPELWRLTVASHHIRIWSGNQTLET RDRKPFYTVDLNSSLTLPLQSCVKPPYMLVVGNIVIKPDSQTITCENCRLLTCIDSTFNWQHRILLVRAREG VWIPVSMDRPWEASPSVHILTEVLKGVLNRSKRFIFTLIAVIMGLIAVTATAAVAGVALHSSVQSVNFVNDG QKNSTRLWNSQSSIDQKLANQINDLRQTVIWMGDRLMSLEHRFQLQCDWNTSDFCITPQIYNDSEHHWDMVR RHLQGREDNLTLDISKLKEQIFEASKAHLNLVPGTEAIAGVADGLANLNPVTWVKTIGSTTIINLILILVCL FCLLLVCRCTQQLRRDSDHRERAMMTMAVLSKRKGGNVGKSKRDQIVTVSV >AC078899.1 Homo sapiens chromosome 19, BACBC371065 envT-ENVT: (SEQ ID NO: 11) MGPEAWVRPLKTAPKPGEAIRLILFIYLSCFFLPVMSSEPSYSFLLTSFTTGRVFANTTWRAGTSKEVSFAV DLCVLFPEPARTHEEQHNLPVIGAGSVDLAAGFGHSGSQTGCGSSKGAEKGLQNVDFYLCPGNHPDASCRDT YQFFCPDWTCVTLATYSGGSTRSSTLSISRVPHPKLCTRKNCNPLTITVHDPNAAQWYYGMSWGLRLYIPGF DVGTMFTIQKKILVSWSSPKPIGPLTDLGDPIFQKHPDKVDLTVPLPFLVPRPQLQQQHLQPSLMSILGGVH HLLNLTQPKLAQDCWLCLKAKPPYYVGLGVEATLKRGPLSCHTRPRALTIGDVSGNASCLISTGYNLSASPF QATCNQSLLTSISTSVSYQAPNNTWLACTSGLTRCINGTEPGPLLCVLVHVLPQVYVYSGPEGRQLIAPPEL HPRLHQAVPLLVPLLAGLSIAGSAAIGTAALVQGETGLISLSQQVDADFSNLQSAIDILHSQVESLAEVVLQ NCRCLDLLFLSQGGLCAALGESCCFYANQSGVIKGTVKKVRENLDRHQQERENNIPVVYQSMFNWNPWLTTL ITGLAGPLLILLLSLIFGPCILNSFLNFIKQRIASVKLTYLKTQYDTLVNN >AC000064.1 Human BAC clone RG083M05 from 7q21-7q22 envW (Syncytin-1)- ENVW (Syncytin-1): (SEQ ID NO: 12) MALPYH1FLFTVLLPSFTLTAPPPCRCMTSSSPYQEFLWRMQRPGNIDAPSYRSLSKGTPTFTAHTHMPRNC YHSATLCMHANTHYWTGKMINPSCPGGLGVTVCWTYFTQTGMSDGGGVQDQAREKHVKEVISQLTRVHGTSS PYKGLDLSKLHETLRTHTRLVSLFNTTLTGLHEVSAQNPTNCWICLPLNFRPYVSIPVPEQWNNFSTEINTT SVLVGPLVSNLEITHTSNLTCVKFSNTTYTTNSQCIRWTPPTQIVCLPSGIFFVCGTSAYRCLNGSSESMCF LSFLVPPMTIYTEQDLYSYVISKPRNKRVPILPFVIGAGVLGALGTGIGGITTSTQFYYKLSQELNGDMERV ADSLVTLQDQLNSLAAVVLQNRRALDLLTAERGGTCLFLGEECCYYVNQSGIVTEKVKEIRDRIQRRAEELR NTGPWGLLSQWMPWILPFLGPLAAIILLLLFGPCIFNLLVNFVSSRIEAVKLQMEPKMQSKTKIYRRPLDRP ASPRSDVNDIKGTPPEEISAAQPLLRPNSAGSS >AL136139.6 Human DNA sequence from clone RP4-761I2 envFRD-ENVFRD (Syncytin-2): (SEQ ID NO: 13) MGLLLLVLILTPSLAAYRHPDFPLLEKAQQLLQSTGSPYSTNCWLCTSSSTETPGTAYPASPREVVTSIEAE
LHISYRWDPNLKGLMRPANSLLSTVKQDFPDIRQKPPIFGPIFTNINLMGIAPICVMAKRKNGTNVGTLPST VCNVTFTVDSNQQTYQTYTHNQFRHQPRFPKPPNITFPQGTLLDKSSRFCQGRPSSCSTRNFWFRPADYNQC LQISNLSSTAEWVLLDQTRNSLFWENKTKGANQSQTPCVQVLAGMTIATSYLGISAVSEFFGTSLTPLFHFH ISTCLKTQGAFYICGQSIHQCLPSNVVTGTCTIGYVTPDIFIAPGNLSLPIPIYGNSPLPRVRRAIHFIPLL AGLGILAGTGTGIAGITKASLTYSQLSKEIANNIDTMAKALTTMQEQIDSLAAVVLQNRRGLDMLTAAQGGI CLALDEKCCFWVNQSGKVQDNIRQLLNQASSLRERATQGWLNWEGTWKWFSWVLPLTGPLVSLLLLLLFGPC LLNLITQFVSSRLQAIKLQTNLSAGRHPRNIQESPF >AC073210.8 Homo sapiens BAC clone RP11-460N20 envR-ENVR: (SEQ ID NO: 14) MLGMNMLLITLFLLLPLSMLKGEPWEGCLHCTHTTWSGNIMTKTLLYHTYYECAGTCLGTCTHNQTTYSVCD PGRGQPYVCYDPKSSPGTWFEIHVGSKEGDLLNQTKVFPSGKDVVSLYFDVCQIVSMGSLFPVIFSSMEYYS SCHKNRYAHPACSTDSPVTTCWDCTTWSTNQQSLGPIMLTKIPLEPDCKTSTCNSVNLTILEPDQPIWTTGL KAPLGARVSGEEIGPGAYVYLYIIKKTRTRSTQQFRVFESFYEHVNQKLPEPPPLASNLFAQLAENIASSLH VASCYVCGGMNMGDQWPWEARELMPQDNFTLTASSLEPAPSSQSIWFLKTSIIGKFCIARWGKAFTDPVGEL TCLGQQYYNETLGKTLWRGKSNNSESPHPSPFSRFPSLNHSWYQLEAPNTWQAPSGLYWICGPQAYRQLPAK WSGACVLGTIRPSFFLMPLKQGEALGYPIYDETKRKSKRGITIGDWKDNEWPPERIIQYYGPATWAEDGMWG YRTPVYMLNRIIRLQAVLEIITNETAGALNLLAQQATKMRNVIYQNRLALDYLLAQEEGVCGKFNLTNCCLE LDDEGKVIKEITAKIQKLAHIPVQTWKG >AC093488.1 Homo sapiens chromosome 3 clone RP11-10O8 envR(b)-ENVR(b): (SEQ ID NO: 15) MDPLHTIEKVPARRNIHDRGHQGHRMGDGTPGRPKISVQQMTRFSLIIFFLSAPFVVNASTSNVFLQWAHSY ADGLQQGDPCWVCGSLPVTNTMELPWWVSPLQGKDWVFFQSFIGDLKQVVTGAQMTGVTRKNISEWPINKTL NEPGHDKPFSVNETRDKVIAFAIPLLDTKVFVQTSRPQNTQYRNGFLQIWDGFIWLTATKGHLSQIAPLCWE QRNHSLDNWPNTTRVMGWIPPGQCRHTILLQQRDLFATDWSQQPGLNWYAPNGTQWLCSPNLWPWLPSGWLG CCTLGIPWAQGRWVKTMEVYPYLPHVVNQGTRAIVHRNDHLPTIFMPSVGLGTVIQHIEALANFTQRALNDS LQSISLMNAEVYYMHEDILQNRMALDILTAAEGGTCALIKTECCVYIPNNSRNISLALEDTCRQIQVISSSA LSLHDWIASQFSGRPSWWQKILIVLATLWSVGIALCCGLYFCRMFSQHIPQTHSIIFQQELPLSPPSQEHYQ SQRDIFHSNAP >AC016222.4 Homo sapiens clone RP11-26J6 envF(c)2-ENVF(c)2: (SEQ ID NO: 16) MNSPCDRLQQFIQVLLEESWSFPSFANTLHWPENLLSYIDELVWQGSLQNFHQHEVRFDKPPLRLPLTGFSS LTENWSSRQAVSSRLVATAASPPAGCQAPIAFLGLKFSSLGPARKNPALCFLYDQSNSKCNTSWVKENVGCP WHWCNIHEALIRTEKGSDPMFYVNTSTGGRDGFNGFNLQISDPWDPRWASGVDGGLYEHKTFMYPVAKIRIA RTLKTTVTGLSDLASSIQSAEKELTSQLQPAADQAKSSRFSWLTLISEGAQLLQSTGVQNLSHCFLCAALRR PPLVAVPLPTPFNYTINSSTPIPPVPKGQVPLFSDPIRHKFPFCYSTPNASWCNQTRMLTSTPAPPRGYFWC NSTLTKVLNSTGNHTLCLPISLIPGLTLYSQDELSHLLAWTEPRPQNKSKWAIFLPLVLGISLASSLVASGL GKGALTHSIQTSQDLSTHLQLAIEASAESLDSLQRQITTVAQVAAQNRQALDLLMAEKGRTCLFLQEECCYY LNESGVVENSLQTLKKKKSSKRS >AL354685.17 Human DNA sequence from clone RP13-75G22 envF(c)1-ENVF(c)1: (SEQ ID NO: 17) MARPSPLCLLLLLTLLTPIVPSNSLLTEPPFRWRFYLHETWTQGNRLSTVTLATVDCQPHGCQAQVTFNFTS FKSVLRGWSNPTICFVYDQTHSNCRDYWVDTNGGCPYAYCRMHVTQLHTAKKLQHTYRLTSDGRTTYFLTIP DPWDSRWVSGVTGRLYRWPTDSYPVGKLRIFLTYIRVIPQVLSNLKDQADNIKHQEEVINTLVQSHPKADMV TYDDKAEAGPFSWITLVRHGARLVNMAGLVNLSHCFLCTALSQPPLVAVPLPQAFNTSGNHTAHPSGVFSEQ VPLFRDPLQPQFPFCYTTPNSSWCNQTYSGSLSNLSAPAGGYFWCNFTLTKHLNISSNNTLSRNLCLPISLV PRLTLYSEAELSSLVNPPMRQKRAVFPPLVIGVSLTSSLVASGLGTGAIVHFISSSQDLSIKLQMAIEASAE SLASLQRQITSVAKVAMQNRRALDLLTADKGGTCMFLGEECCYYINESGLVETSLLTLDKIRDGLHRPSSTP NYGGGVWVQSPLTTWIIPFISPILIICLLLLIAPCVLKFIKNRISEVSRVTVNQMLLHPYSRLPTSEDHYDD ALTQQEAAR HERV-Kcon ENV-hENVKcon: (SEQ ID NO: 18) MNPSEMQRKAPPRRRRHRNRAPLTHKMNKMVTSEEQMKLPSTKKAEPPTWAQLKKLTQLATKYLENTKVTQT PESMLLAALMIVSMVVSLPMPAGAAAANYTYWAYVPFPPLIRAVTWMDNPIEVYVNDSVWVPGPIDDRCPAK PEEEGMMINISIGYRYPPICLGRAPGCLMPAVQNWLVEVPTVSPISRFTYHMVSGMSLRPRVNYLQDFSYQR SLKFRPKGKPCPKEIPKESKNTEVLVWEECVANSAVILQNNEFGTIIDWAPRGQFYHNCSGQTQSCPSAQVS PAVDSDLTESLDKHKHKKLQSFYPWEWGEKGISTPRPKIVSPVSGPEHPELWRLTVASHHIRIWSGNQTLET RDRKPFYTVDLNSSLTVPLQSCVKPPYMLVVGNIVIKPDSQTITCENCRLLTCIDSTFNWQHRILLVRAREG VWIPVSMDRPWEASPSVHILTEVLKGVLNRSKRFIFTLIAVIMGLIAVTATAAVAGVALHSSVQSVNFVNDW QKNSTRLWNSQSSIDQKLANQINDLRQTVIWMGDRLMSLEHRFQLQCDWNTSDFCITPQIYNESEHHWDMVR RHLQGREDNLTLDISKLKEQIFEASKAHLNLVPGTEAIAGVADGLANLNPVTWVKTIGSTTIINLILILVCL FCLLLVCRCTQQLRRDSDHRERAMMTMAVLSKRKGGNVGKSKRDQIVTVSV HERV-Kcon GAG-hGAGKcon: (SEQ ID NO: 19) MGQTKSKIKSKYASYLSFIKILLKRGGVKVSTKNLIKLFQIIEQFCPWFPEQGTLDLKDWKRIGKELKQAGR KGNIIPLTVVVNDWAIIKAALEPFQTEEDSVSVSDAPGSCIIDONENTRKKSQKETEGLHCEYVAEPVMAQS TQNVDYNQLQEVIYPETLKLEGKGPELVGPSESKPRGTSPLPAGQVPVTLQPQKQVKENKTQPPVAYQYWPP AELQYRPPPESQYGYPGMPPAPQGRAPYPQPPTRRLNPTAPPSRQGSELHEIIDKSRKEGDTEAWQFPVTLE PMPPGEGAQEGEPPTVEARYKSFSIKMLKDMKEGVKQYGPNSPYMRTLLDSIAHGHRLIPYDWEILAKSSLS PSQFLQFKTVWVIDGVQEQVRRNRAANPPVNIDADQLLGIGQNWSTISQQALMQNEAIEQVRAICLRAWEKI QDPGSTCPSFNTVRQGSKEPYPDFVARLQDVAQKSIADEKARKVIVELMAYENANPECQSAIKPLKGKVPAG SDVISEYVKACDGIGGAMHKAMLMAQAITGVVLGGQVRTFGGKCYNCGQIGHLKKNCPVLNKQNITIQATTT GREPPDLCPRCKKGKHWASQCRSKFDKNGQPLSGNEQRGQPQAPQQTGAFPIQPFVPQGFQGQQPPLSQVFQ GISQLPQYNNCPPPQAAVQQ Rattus norvegicus & synthetic: APOBEC1-XTENL8-nspCas9-UGI-SV40 NLS (SEQ ID NO: 20) MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTT ERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIM TEQESGYCWRNFVNYSPSNEAHWPRYPHLVVVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHY QRLPPHILWATGLKSGSETPGTSESATPESDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSI KKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHER HPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQ LVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLA EDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQD LTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQR TFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITP WNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKA IVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDI VLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANR NFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEM ARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSD YDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGG LSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREIN NYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEIT LANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARK KDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLI IKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLD EIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTST KEVLDATLIHQSITGLYETRIDLSQLGGDSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESD ILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKMLSGGSPKKKRKV Streptococcus pyogenes: spCas9 Bipartite NLS (SEQ ID NO: 21) MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRY TRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVD STDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARL SKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYAD LFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYA GYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPF LKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPN EKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVM KQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSL HEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELG SQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNR GKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILD SRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEF VYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDF ATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKG KSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGN ELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNK HRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD GSGGGGSGKRTADGSEFEPKKKRKVSSGGDYKDHDGDYKDHDIDYKDDDDK Staphylococcus aureus: saCas9 (SEQ ID NO: 22) MKRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEGRRSKRGARRLKRRRRHRIQRVKKLLF DYNLLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKA LEEKYVAELQLERLKKDGEVRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPG EGSPFGWKDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQIIENVF KQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEIIENAELLDQIAKILTIYQS SEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAINLILDELWHTNDNQIAIFNRLKLVPKKVDLSQ QKEIPTTLVDDFILSPVVKRSFIQSIKVINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNER IEEIIRTTGKENAKYLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVK QEENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFSVQKDFINRNLVD TRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERNKGYKHHAEDALIIANADFIFKEWKK LDKAKKVMENQMFEEKQAESMPEIETEQEYKEIFITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTR
KDDKGNTLIVNNLNGLYDKDNDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNY LTKYSKKDNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKK ENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLLNRIEVNMIDITYREYLENM NDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQIIKKG Acidaminococcus sp.: asCas12a (SEQ ID NO: 23) MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDHYKELKPIIDRIYKTYADQCLQLVQLD WENLSAAIDSYRKEKTEETRNALIEEQATYRNAIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLK QLGTVTTTEHENALLRSFDKFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVP SLREHFENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLNLAIQKN DETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLRNENVLETAEALFNELNSID LTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGKITKSAKEKVQRSLKHEDINLQEIISAAGKELS EAFKQKTSEILSHAHAALDQPLPTTLKKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIK LEMEPSLSFYNKARNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYK ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPLEITKEIYDLNNP EKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDLSSLRPSSQYKDLGEYYAELNPLLYH ISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGHHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPK SRMKRMAHRLGEKMLNKKLKDQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRF TSDKFFFHVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRSLNTIQ QFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVVVLENLNFGFKSKRTGIAE KAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQFTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFV DPFVWKTIKNHESRKHFLEGFDFLHYDVKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGT PFIAGKRIVPVIENHRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQ MRNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESKDLKLQNGISNQ DWLAYIQELRN Pleckstrin homology domain of Homo sapiens phospholipase C81 (hPLC81) (SEQ ID NO: 24) MDSGRDFLTLHGLQDDEDLQALLKGSQLLKVKSSSWRRERFYKLQEDCKTIWQESRKVMRTPESQLFSIEDI QEVRMGHRTEGLEKFARDVPEDRCFSIVFKDQRNTLDLIAPSPADAQHWVLGLHKIIHHSGSMDQRQKLQHW IHSCLRKADKNKDNKMSFKELQNFLKELNIQ Pleckstrin homology domain of Homo sapiens Akt1 (hAkt) (SEQ ID NO: 25) MSDVAIVKEGWLHKRGEYIKTWRPRYFLLKNDGTFIGYKERPQDVDQREAPLNNFSVAQCQLMKTERPRPNT FIIRCLQWTTVIERTFHVETPEEREEWTTAIQTVADGLKKQEEEEMDFRSGSPSDNSGAEEMEVSLAKPKHR VTMNEFEYLKLLGKGTFGKVDPPV Pleckstrin homology domain of Homo sapiens PDPK1 (hPDPK1) (SEQ ID NO: 26) KMGPVDKRKGLFARRRQLLLTEGPHLYYVDPVNKVLKGEIPWSQELRPEAKNFKTFFVHTPNRTYYLMDPSG NAHKWCRKIQEVWRQRYQSH Homo sapiens: CD9 Complete Protein (SEQ ID NO: 27) MSPVKGGTKCIKYLLFGFNFIFWLAGIAVLAIGLWLRFDSQTKSIFEQETNNNNSSFYTGVYILIGAGALMM LVGFLGCCGAVQESQCMLGLFFGFLLVIFAIEIAAAIWGYSHKDEVIKEVQEFYKDTYNKLKTKDEPQRETL KAIHYALNCCGLAGGVEQFISDICPKKDVLETFTVKSCPDAIKEVFDNKFHIIGAVGIGIAVVMIFGMIFSM ILCCAIRRNREMV Homo sapiens: C063 Complete Protein (SEQ ID NO: 28) MAVEGGMKCVKFLLYVLLLAFCACAVGLIAVGVGAQLVLSQTIIQGATPGSLLPVVIIAVGVFLFLVAFVGC CGACKENYCLMITFAIFLSLIMLVEVAAAIAGYVFRDKVMSEFNNNFRQQMENYPKNNHTASILDRMQADFK CCGAANYTDWEKIPSMSKNRVPDSCCINVTVGCGINFNEKAIHKEGCVEKIGGWLRKNVLVVAAAALGIAFV EVLGIVFACCLVKSIRSGYEVM Homo sapiens: CD81 Complete Protein (SEQ ID NO: 29) MGVEGCTKCIKYLLFVFNFVFWLAGGVILGVALWLRHDPQTTNLLYLELGDKPAPNTFYVGIYILIAVGAVM MFVGFLGCYGAIQESQCLLGTFFTCLVILFACEVAAGIWGFVNKDQIAKDVKQFYDQALQQAVVDDDANNAK AVVKTFHETLDCCGSSTLTALTTSVLKNNLCPSGSNIISNLFKEDCHQKIDDLFSGKLYLIGIAAIVVAVIM IFEMILSMVLCCGIRNSSVY Homo sapiens: CD47 "Self Hairpin" 10 Amino Acids (SEQ ID NO: 30) EVTELTREGE Homo sapiens: CD47 "Self Hairpin" 21 Amino Acids (SEQ ID NO: 31) GNYTCEVTELTREGETIIELK Homo sapiens: CD47 Complete Protein (SEQ ID NO: 32) MWPLVAALLLGSACCGSAQLLFNKTKSVEFTFCNDTVVIPCFVTNMEAQNTTEVYVKWKFKGRDIYTFDGAL NKSTVPTDFSSAKIEVSQLLKGDASLKMDKSDAVSHTGNYTCEVTELTREGETIIELKYRVVSWFSPNENIL IVIFPIFAILLFWGQFGIKTLKYRSGGMDEKTIALLVAGLVITVIVIVGAILFVPGEYSLKNATGLGLIVTS TGILILLHYYVFSTAIGLTSFVIAILVIQVIAYILAVVGLSLCIAACIPMHGPLLISGLSILALAQLLGLVY MKFVE Synthetic: dDZF1 (SEQ ID NO: 33) FKCEHCRILFLDHVMFTIHMGCHGFRDPFKCNMCGEKCDGPVGLFVHMARNAHGEKPFYCEHCEITFRDVVM YSLHKGYHGFRDPFECNICGYHSQDRYEFSSHIVRGEH Synthetic: dDZF2 (SEQ ID NO: 34) HHCQHCDMYFADNILYTIHMGCHSCDDVFKCNMCGEKCDGPVGLFVHMARNAHGEKPTKCVHCGIVFLDEVM YALHMSCHGFRDPFECNICGYHSQDRYEFSSHIVRGEH Synthetic: DmrA (SEQ ID NO: 35) MGRGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKPFKFMLGKQEVIRGWEEGVAQMSVG QRAKLTISPDYAYGATGHPGIIPPHATLVFDVELLKLE Synthetic: DmrB (SEQ ID NO: 36) MASRGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKPFKFMLGKQEVIRGWEEGVAQMSV GQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELLKLE Synthetic: DmrC (SEQ ID NO: 37) MGSRILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETSFNQAYGRDLMEAQEWCRKY MKSGNVKDLLQAWDLYYHVFRRISK Homo sapiens/Synthetic: FKBP (SEQ ID NO: 38) MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKPFKFMLGKQEVIRGWEEGVAQMSVGQR AKLTISPDYAYGATGHPGIIPPHATLVFDVELLKLE Homo sapiens/Synthetic: FRB (SEQ ID NO: 39) QGMLEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETSFNQAYGRDLMEAQEWCRKYMKSG NVKDLLQAWDLYYHVFRRISK Synthetic: Anti-GCN4 scFv (SEQ ID NO: 40) MGPDIVMTQSPSSLSASVGDRVTITCRSSTGAVTTSNYASWVQEKPGKLFKGLIGGTNNRAPGVPSRFSGSL IGDKATLTISSLQPEDFATYFCALWYSNHVVVFGQGTKVELKRGGGGSGGGGSGGGGSSGGGSEVKLLESGG GLVQPGGSLKLSCAVSGFSLTDYGVNVVVRQAPGRGLEWIGVIWGDGITDYNSALKDRFIISKDNGKNTVYL QMSKVRSDDTALYYCVTGLFDYWGQGTLVTVSSYPYDVPDYAGGGGGSGGGGSGGGGSGGGGS Synthetic: 10x-GCN4 Repeats (SEQ ID NO: 41) EELLSKNYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGSGSG EELLSKNYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGSGSG EELLSKNYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGSGSG EELLSKNYHLENEVARLKKGS Synthetic: 24x-GCN4 Repeats (SEQ ID NO: 42) EELLSKNYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGSGSG EELLSKNYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGSGSG EELLSKNYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGSGSG EELLSKNYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGSGSG EELLSKNYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGSGSG EELLSKNYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGSGSG EELLSKNYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGSGSG EELLSKNYHLENEVARLKKGSGSGEELLSKDYHLENEVARLKKGSGSGEELLSKNYHLENEVARLKKGS Synthetic: GFP-targeting Nanobody (SEQ ID NO: 43) VQLVESGGALVQPGGSLRLSCAASGFPVNRYSMRWYRQAPGKEREWVAGMSSAGDRSSYEDSVKGRFTISRD DARNTVYLQMNSLKPEDTAVYYSNVNVGFEYWGQGTQVTVSS Nostoc punctiforme: Npu DnaE N-terminal Split Intein (SEQ ID NO: 44) CLSYETEILTVEYGLLPIGKIVEKRIECTVYSVDNNGNIYTQPVAQWHDRGEQEVFEYCLEDGSLIRATKDH KFMTVDGQMLPIDEIFERELDLMRVDNLPN Nostoc punctiforme: Npu DnaE C-terminal Split Intein (SEQ ID NO: 45) MIKIATRKYLGKQNVYDIGVERDHNFALKNGFIASNCFN Synthetic: Cfa N-Terminal Split Intein (SEQ ID NO: 46 CLSYDTEILTVEYGFLPIGKIVEERIECTVYTVDKNGFVYTQPIAQWHNRGEQEVFEYCLEDGSIIRATKDH KFMTTDGQMLPIDEIFERGLDLKQVDGLP Synthetic: Cfa C-Terminal Split Intein (SEQ ID NO: 47) MVKIISRKSLGTQNVYDIGVEKDHNFLLKNGLVASN Saccharomyces cerevisiae: Vma N-terminal Split Intein (SEQ ID NO: 48) CFAKGTNVLMADGSIECIENIEVGNKVMGKDGRPREVIKLPRGRETMYSVVQKSQHRAHKSDSSREVPELLK FTCNATHELVVRTPRSVRRLSRTIKGVEYFEVITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELVE SYRKASNKAYFEWTIEARDLSLLGSHVRKATYQTYAPILY Saccharomyces cerevisiae: Vma C-terminal Split Intein (SEQ ID NO: 49) VLLNVLSKCAGSKKFRPAPAAAFARECRGFYFELQELKEDDYYGITLSDDSDHQFLLANQVVVHN Synechocystis sp. PCC 6803: Ssp DnaE N-terminal Split Intein (SEQ ID NO: 50) CLSFGTEILTVEYGPLPIGKIVSEEINCSVYSVDPEGRVYTQAIAQWHDRGEQEVLEYELEDGSVIRATSDH RFLTTDYQLLAIEEIFARQLDLLTLENIKQTEEALDNHRLPFPLLDAGTIK Synechocystis sp. PCC 6803: Ssp DnaE C-terminal Split Intein (SEQ ID NO: 51) MVKVIGRRSLGVQRIFDIGLPQDHNFLLANGAIAAN Synthetic: Spy Tag (SEQ ID NO: 52) VPTIVMVDAYKRYK
Synthetic: Spy Catcher (SEQ ID NO: 53) MVTTLSGLSGEQGPSGDMTTEEDSATHIKFSKRDEDGRELAGATMELRDSSGKTISTWISDGHVKDFYLYPG KYTFVETAAPDGYEVATAITFTVNEQGQVTVNGEATKGDAHTGSSGS Bacteriophage A452: MS2 RNA Binding Protein (SEQ ID NO: 54) MASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVTCSVRQSSAQNRKYTIKVEVPKVATQT VGGVELPVAAWRSYLNMELTIPIFATNSDCELIVKAMQGLLKDGNPIPSAIAANSGIY Bacteriophage A452: MS2 (N55K) RNA Binding Protein (SEQ ID NO: 55) MASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVTCSVRQSSAQKRKYTIKVEVPKVATQT VGGVELPVAAWRSYLNMELTIPIFATNSDCELIVKAMQGLLKDGNPIPSAIAANSGIY Bacteriophage A452: MS2 (N55K)(V29I) RNA Binding Protein (SEQ ID NO: 56) MASNFTQFVLVDNGGTGDVTVAPSNFANGIAEWISSNSRSQAYKVTCSVRQSSAQKRKYTIKVEVPKVATQT VGGVELPVAAWRSYLNMELTIPIFATNSDCELIVKAMQGLLKDGNPIPSAIAANSGIY Bacteriophage PP7: PP7 RNA Binding Protein (SEQ ID NO: 57) KTIVLSVGEATRTLTEIQSTADRQIFEEKVGPLVGRLRLTASLRQNGAKTAYRVNLKLDQADVVDSGLPKVR YTQVWSHDVTIVANSTEASRKSLYDLTKSLVATSQVEDLVVNLVPLGRS Bacteriophage Mu: COM RNA Binding Protein (SEQ ID NO: 58) MKSIRCKNCNKLLFKADSFDHIEIRCPRCKRHIIMLNACEHPTEKHCGKREKITHSDETVRY Synthetic: Zinc Finger ZF6/10 (SEQ ID NO: 59) STRPGERPFQCRICMRNFSIPNHLARHTRTHTGEKPFQCRICMRNFSQSAHLKRHLRTHTGEKPFQCRICMR NFSQDVSLVRHLKTHLRQKDGERPFQCRICMRNFSSAQALARHTRTHTGEKPFQCRICMRNFSQGGNLTRHL RTHTGEKPFQCRICMRNFSQHPNLTRHLKTHLRGS Synthetic: Zinc Finger ZF8/7 (SEQ ID NO: 60) SRPGERPFQCRICMRNFSTMAVLRRHTRTHTGEKPFQCRICMRNFSRREVLENHLRTHTGEKPFQCRICMRN FSQTVNLDRHLKTHLRQKDGERPFQCRICMRNFSKKDHLHRHTRTHTGEKPFQCRICMRNFSQRPHLTNHLR THTGEKPFQCRICMRNFSVGASLKRHLKTHLRGS Synthetic: Zinc Finger ZF9 (SEQ ID NO: 61) SRPGERPFQCRICMRNFSDKTKLRVHTRTHTGEKPFQCRICMRNFSVRHNLTRHLRTHTGEKPFQCRICMRN FSQSTSLQRHLKTHLRGF Synthetic: Zinc Finger MK10 (SEQ ID NO: 62) SRPGERPFQCRICMRNFSRRHGLDRHTRTHTGEKPFQCRICMRNFSDHSSLKRHLRTHTGSQKPFQCRICMR NFSVRHNLTRHLRTHTGEKPFQCRICMRNFSDHSNLSRHLKTHTGSQKPFQCRICMRNFSQRSSLVRHLRTH TGEKPFQCRICMRNFSESGHLKRHLRTHLRGS Synthetic: FokI Zinc Finger Nuclease 17-2 Targeting GFP (SEQ ID NO: 63) SRPGERPFQCRICMRNFSTRQNLDTHTRTHTGEKPFQCRICMRNFSRRDTLERHLRTHTGEKPFQCRICMRN FSRPDALPRHLKTHLRGSQLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKVMEFFMKVYG YRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQRYVEENQTRNKHINPNEWWKVYP SSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVLSVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF Synthetic: FokI Zinc Finger Nuclease 18-2 Targeting GFP (SEQ ID NO: 64) SRPGERPFQCRICMRNFSSPSKLIRHTRTHTGEKPFQCRICMRNFSDGSNLARHLRTHTGEKPFQCRICMRN FSRVDNLPRHLKTHLRGSQLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKVMEFFMKVYG YRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQRYVEENQTRNKHINPNEWWKVYP SSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVLSVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF Synthetic: FokI Nuclease Domain (SEQ ID NO: 65) QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKVMEFFMKVYGYRGKHLGGSRKPDGAIYT VGSPIDYGVIVDTKAYSGGYNLPIGQADEMQRYVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGN YKAQLTRLNHITNCNGAVLSVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF Synthetic: AcuI Nuclease Domain (SEQ ID NO: 66) VHDHKLELAKLIRNYETNRKECLNSRYNETLLRSDYLDPFFELLGWDIKNKAGKPTNEREVVLEEALKASAS EHSKKPDYTFRLFSERKFFLEAKKPSVHIESDNETAKQVRRYGFTAKLKISVLSNFEYLVIYDTSVKVDGDD TFNKARIKKYHYTEYETHFDEICDLLGRESVYSGNFDKEWLSIENKINHFSVDTL Synthetic: Truncated AcuI Nuclease Domain (SEQ ID NO: 67) YNETLLRSDYLDPFFELLGWDIKNKAGKPTNEREVVLEEALKASASEHSKKPDYTFRLFSERKFFLEAKKPS VHIESDNETAKQVRRYGFTAKLKISVLSNFEYLVIYDTSVKVDGDDT Ruminococcus flavefaciens: RfxCas13d (CasRx) (SEQ ID NO: 68) EASIEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDSIRSVNEGEAFSAEMADKNAGYKIGN AKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLEKRYFGESADGNDNICIQVIHNILDIEKILAEYITNAAY AVNNISGLDKDIIGFGKFSTVYTYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFS KEGRNYIINYGNECYDILALLSGLRHVVVVHNNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITNELTN SFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRKDMSEIRKNHKVFDSIRT KVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFVINLRGSFNDDQKDALYYDEANRIWRKLENI MHNIKEFRGNKTREYKKKDAPRLPRILPAGRDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKV MPLIGVNAKFVEEYAFFKDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFS LDENGNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQKKQGQNGKNQ IDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGRENAEREKFKKIISLYLTVIYHILK NIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLEEKGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKES IDSLESANPKLYANYIKYSDEKKAEEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFRNKA VHLEVARYVHAYINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCVPFG YCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNSGSG Ruminococcus flavefaciens & Synthetic: dead RfxCas13d (dCasRx) (SEQ ID NO: 69) EASIEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDSIRSVNEGEAFSAEMADKNAGYKIGN AKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLEKRYFGESADGNDNICIQVIHNILDIEKILAEYITNAAY AVNNISGLDKDIIGFGKFSTVYTYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFS KEGRNYIINYGNECYDILALLSGLAHWVVANNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITNELTNS FSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRKDMSEIKNHKVFDSIRTKV YTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFVINLRGSFNDDQKDALYYDEANRIWRKLENIMH NIKEFRGNKTREYKKKDAPRLPRILPAGRDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMP LIGVNAKFVEEYAFFKDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLD ENGNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQKKQGQNGKNQID RYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGRENAEREKFKKIISLYLTVIYHILKNI VNINARYVIGFHCVERDAQLYKEKGYDINLKKLEEKGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESID SLESANPKLYANYIKYSDEKKAEEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFANKAVA LEVARYVHAYINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCVPFGYC IPRFKNLSIEALFDRNEAAKFDKEKKKVSGNSGSGPKKKRKVAAAYPYDVPDYA Synthetic: L17E (SEQ ID NO: 70) IWLTALKFLGKHAAKHEAKQQLSKL Synthetic: L17E-Transmembrane (SEQ ID NO: 71) IWLTALKFLGKHAAKHEAKQQLSKLNAVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQK KPR Synthetic: KALA (SEQ ID NO: 72) WEAKLAKALAKALAKHLAKALAKALKACEA Synthetic: KALA-Transmembrane (SEQ ID NO: 73) WEAKLAKALAKALAKHLAKALAKALKACEANAVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILI MLWQKKPR Synthetic: Vectofusin (SEQ ID NO: 74) KKALLHAALAHLLALAHHLLALLKKA Synthetic: Vectofusin-Transmembrane (SEQ ID NO: 75) KKALLHAALAHLLALAHHLLALLKKANAVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ KKPR Synthetic: Transmembrane Domain (SEQ ID NO: 76) NAVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQKKPR Lactococcus lactis: Nisin A (SEQ ID NO: 77) ITSISLCTPGCKTGALMGCNMKTATCHCSIHVSK Lactococcus lactis NIZO 22186: Nisin Z (SEQ ID NO: 78) ITSISLCTPGCKTGALMGCNMKTATCNCSIHVSK Lactococcus lactis subsp. lactis F10: Nisin F (SEQ ID NO: 79) ITSISLCTPGCKTGALMGCNMKTATCNCSVHVSK Lactococcus lactis 61-14: Nisin Q (SEQ ID NO: 80) ITSISLCTPGCKTGVLMGCNLKTATCNCSVHVSK Streptococcus hyointestinalis: Nisin H (SEQ ID NO: 81) FTSISMCTPGCKTGALMTCNYKTATCHCSIKVSK Streptococcus uberis: Nisin U (SEQ ID NO: 82) ITSKSLCTPGCKTGILMTCPLKTATCGCHFG Streptococcus uberis: Nisin U2 (SEQ ID NO: 83) VTSKSLCTPGCKTGILMTCPLKTATCGCHFG Streptococcus galloyticus subsp. pasteurianus: Nisin P (SEQ ID NO: 84) VTSKSLCTPGCKTGILMTCAIKTATCGCHFG L. lactis NZ9800: Nisin A S29A (SEQ ID NO: 85) ITSISLCTPGCKTGALMGCNMKTATCHCAIHVSK L. lactis NZ9800: Nisin A S29D (SEQ ID NO: 86) ITSISLCTPGCKTGALMGCNMKTATCHCDIHVSK L. lactis NZ9800: Nisin A S29E (SEQ ID NO: 87) ITSISLCTPGCKTGALMGCNMKTATCHCEIHVSK L. lactis NZ9800: Nisin A S29G (SEQ ID NO: 88) ITSISLCTPGCKTGALMGCNMKTATCHCGIHVSK L. lactis NZ9800: Nisin A K22T (SEQ ID NO: 89)
ITSISLCTPGCKTGALMGCNMTTATCHCSIHVSK L. lactis NZ9800: Nisin A N20P (SEQ ID NO: 90) ITSISLCTPGCKTGALMGCPMKTATCHCSIHVSK L. lactis NZ9800: Nisin A M21V (SEQ ID NO: 91) ITSISLCTPGCKTGALMGCNVKTATCHCSIHVSK L. lactis NZ9800: Nisin A K22S (SEQ ID NO: 92) ITSISLCTPGCKTGALMGCNMSTATCHCSIHVSK L. lactis NZ9800: Nisin Z N20K (SEQ ID NO: 93) ITSISLCTPGCKTGALMGCKMKTATCNCSIHVSK L. lactis NZ9800: Nisin Z M21K (SEQ ID NO: 94) ITSISLCTPGCKTGALMGCNKKTATCNCSIHVSK AAV2: REP52 (SEQ ID NO: 95) MELVGWLVDKGITSEKQW1QEDQASYISFNAASNSRSQIKAALDNAGKIMSLTKTAPDYLVGQQPVEDISSN RIYKILELNGYDPQYAASVFLGWATKKFGKRNTIWLFGPATTGKTNIAEAIAHTVPFYGCVNVVTNENFPFN DCVDKMVIVWVEEGKMTAKVVESAKAILGGSKVRVDQKCKSSAQIDPTPVIVTSNTNMCAVIDGNSTTFEHQ QPLQDRMFKFELTRRLDHDFGKVTKQEVKDFFRWAKDHVVEVEHEFYVKKGGAKKRPAPSDADISEPKRVRE SVAQPSTSDAEASINYADRYQNKCSRHVGMNLMLFPCRQCERMNQNSNICFTHGQKDCLECFPVSESQPVSV VKKAYQKLCYIHHIMGKVPDACTACDLVNVDLDDCIFEQ AAV2: REP78 (SEQ ID NO: 96) MPGFYEIVIKVPSDLDEHLPGISDSFVNWVAEKEWELPPDSDMDLNLIEQAPLTVAEKLQRDFLTEWRRVSK APEALFFVQFEKGESYFHMHVLVETTGVKSMVLGRFLSQIREKLIQRIYRGIEPTLPNWFAVTKTRNGAGGG NKVVDECYIPNYLLPKTQPELQWAWTNMEQYLSACLNLTERKRLVAQHLTHVSQTQEQNKENQNPNSDAPVI RSKTSARYMELVGWLVDKGITSEKQWIQEDQASYISFNAASNSRSQIKAALDNAGKIMSLTKTAPDYLVGQQ PVEDISSNRIYKILELNGYDPQYAASVFLGWATKKFGKRNTIWLFGPATTGKTNIAEAIAHTVPFYGCVNVV TNENFPFNDCVDKMVIVWVEEGKMTAKVVESAKAILGGSKVRVDQKCKSSAQIDPTPVIVTSNTNMCAVIDG NSTTFEHQQPLQDRMFKFELTRRLDHDFGKVTKQEVKDFFRWAKDHVVEVEHEFYVKKGGAKKRPAPSDADI SEPKRVRESVAQPSTSDAEASINYADRYQNKCSRHVGMNLMLFPCRQCERMNQNSNICFTHGQKDCLECFPV SESQPVSVVKKAYQKLCYIHHIMGKVPDACTACDLVNVDLDDCIFEQ AAV2: VP1 (SEQ ID NO: 97) MAADGYLPDWLEDTLSEGIRQVWVKLKPGPPPPKPAERHKDDSRGLVLPGYKYLGPFNGLDKGEPVNEADAA ALEHDKAYDRQLDSGDNPYLKYNHADAEFQERLKEDTSFGGNLGRAVFQAKKRVLEPLGLVEEPVKTAPGKK RPVEHSPVEPDSSSGTGKAGQQPARKRLNFGQTGDADSVPDPQPLGQPPAAPSGLGTNTMATGSGAPMADNN EGADGVGNSSGNWHCDSTWMGDRVITTSTRTWALPTYNNHLYKQISSQSGASNDNHYFGYSTPWGYFDFNRF HCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTQNDGTTTIANNLTSTVQVFTDSEYQLPYVLGSAHQ GCLPPFPADVFMVPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGNNFTFSYTFEDVPFHSSYAHSQSLD RLMNPLIDQYLYYLSRTNTPSGTTTQSRLQFSQAGASDIRDQSRNWLPGPCYRQQRVSKTSADNNNSEYSVV TGATKYHLNGRDSLVNPGPAMASHKDDEEKFFPQSGVLIFGKQGSEKTNVDIEKVMITDEEEIRTTNPVATE QYGSVSTNLQRGNRQAATADVNTQGVLPGMVWQDRDVYLQGPIWAKIPHTDGHFHPSPLMGGFGLKHPPPQI LIKNTPVPANPSTTFSAAKFASFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYNKSVNVDFTVDTNG VYSEPRPIGTRYLTRNL AAV2: VP2 (SEQ ID NO: 98) APGKKRPVEHSPVEPDSSSGTGKAGQQPARKRLNFGQTGDADSVPDPQPLGQPPAAPSGLGTNTMATGSGAP MADNNEGADGVGNSSGNWHCDSTWMGDRVITTSTRTWALPTYNNHLYKQISSQSGASNDNHYFGYSTPWGYF DFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTQNDGTTTIANNLTSTVQVFTDSEYQLPYVL GSAHQGCLPPFPADVFMVPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGNNFTFSYTFEDVPFHSSYAH SQSLDRLMNPLIDQYLYYLSRTNTPSGTTTQSRLQFSQAGASDIRDQSRNWLPGPCYRQQRVSKTSADNNNS EYSVVTGATKYHLNGRDSLVNPGPAMASHKDDEEKFFPQSGVLIFGKQGSEKTNVDIEKVMITDEEEIRTTN PVATEQYGSVSTNLQRGNRQAATADVNTQGVLPGMVWQDRDVYLQGPIWAKIPHTDGHFHPSPLMGGFGLKH PPPQILIKNTPVPANPSTTFSAAKFASFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYNKSVNVDFT VDTNGVYSEPRPIGTRYLTRNL AAV2: VP3 (SEQ ID NO: 99) MATGSGAPMADNNEGADGVGNSSGNWHCDSTWMGDRVITTSTRTWALPTYNNHLYKQISSQSGASNDNHYFG YSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTQNDGTTTIANNLTSTVQVFTDS EYQLPYVLGSAHQGCLPPFPADVFMVPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGNNFTFSYTFEDV PFHSSYAHSQSLDRLMNPLIDQYLYYLSRTNTPSGTTTQSRLQFSQAGASDIRDQSRNWLPGPCYRQQRVSK TSADNNNSEYSVVTGATKYHLNGRDSLVNPGPAMASHKDDEEKFFPQSGVLIFGKQGSEKTNVDIEKVMITD EEEIRTTNPVATEQYGSVSTNLQRGNRQAATADVNTQGVLPGMVWQDRDVYLQGPIWAKIPHTDGHFHPSPL MGGFGLKHPPPQILIKNTPVPANPSTTFSAAKFASFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYN KSVNVDFTVDTNGVYSEPRPIGTRYLTRNL Relevant RNA Sequences (5'-3') Synthetic: MS2 Stem Loop spCas9 Scaffold RNA for sgRNA with Terminator Example 1 (SEQ ID NO: 100) GUUUUAGAGCUAGGCCAACAUGAGGAUCACCCAUGUCUGCAGGGCCUAGCAAGUUAAAAUAAGGCUAGUCCG UUAUCAACUUGGCCAACAUGAGGAUCACCCAUGUCUGCAGGGCCAAGUGGCACCGAGUCGGUGCUUUUUUU Synthetic: MS2 Stem Loop spCas9 Scaffold RNA for sgRNA with Terminator Example 2 (SEQ ID NO: 101) GUUUUAGAGCUAGGCCAACAUGAGGAUCACCCAUGUCUGCAGGGCCUAGCAAGUUAAAAUAAGGCUAGUCCG UUAUCAACUUGGCCAACAUGAGGAUCACCCAUGUCUGCAGGGCCAAGUGGCACCGAGUCGGUGCGGGAGCAC AUGAGGAUCACCCAUGUGCGACUCCCACAGUCACUGGGGAGUCUUCCCUUUUUUU Synthetic: MS2 Stem Loop spCas9 Scaffold RNA for sgRNA with Terminator Example 3 (SEQ ID NO: 102) GUUUAAGAGCUAUGCUGGAAACAGCAUAGCAAGUUUAAAUAAGGCUAGUCCGUUAUCAACUUGAAAAAGUGG CACCGAGUCGGUGCGGGAGCACAUGAGGAUCACCCAUGUGCGACUCCCACAGUCACUGGGGAGUCUUCCCUU UUUUU Synthetic: 4xMS2 Stem Loop RNA Scaffold Example (SEQ ID NO: 103) UUCUAGAUCAUCGAAACAUGAGGAUCACCCAUAUCUGCAGUCGACAUCGAAACAUGAGGAUCACCCAUGUCU GCAGUCGACAUCGAAACAUGAGGAUCACCCAUGUCUGCAGUCGACAUCGAAACAUGAGGAUCACCCAUGUCU GCAGUCGACAUCGAAAUCGAUAAGCUUCAGAUCAGAUCCUAG Synthetic: MS2 Stem Loop Example 1 (SEQ ID NO: 104) ACAUGAGGAUCACCCAUGU Synthetic: MS2 Stem Loop Example 2 (SEQ ID NO: 105) ACAUGAGGAUCACCCAUAU Synthetic: MS2 Stem Loop Example 3 (SEQ ID NO: 106) CCACAGUCACUGGG Synthetic: 2xMS2 Stem Loop Example (SEQ ID NO: 107) ACAUGAGGAUCACCCAUGUCUGCAGGGCCUAGCAAGUUAAAAUAAGGCUAGUCCGUUAUCAACUUGGCCAAC AUGAGGAUCACCCAUGU Synthetic: 2xPP7 Stem Loop spCas9 Scaffold RNA for sgRNA with Terminator Example (SEQ ID NO: 108) GUUUUAGAGCUAGGCCGGAGCAGACGAUAUGGCGUCGCUCCGGCCUAGCAAGUUAAAAUAAGGCUAGUCCGU UAUCAACUUGGCCGGAGCAGACGAUAUGGCGUCGCUCCGGCCAAGUGGCACCGAGUCGGUGCUUUUUUU Synthetic: PP7 Stem Loop Example (SEQ ID NO: 109) GCCGGAGCAGACGAUAUGGCGUCGCUCCGGCC Synthetic: COMStem Loop spCas9 Scaffold RNA for sgRNA with Terminator Example (SEQ ID NO: 110) GUUUAAGAGCUAUGCUGGAAACAGCAUAGCAAGUUUAAAUAAGGCUAGUCCGUUAUCAACUUGAAAAAGUGG CACCGAGUCGGUGCCUGAAUGCCUGCGAGCAUCUUUUUUU Synthetic: COM Stem Loop Example (SEQ ID NO: 111) CUGAAUGCCUGCGAGCAUC Relevant DNA Sequences (5'-3') Synthetic: Zinc Finger ZF6/10 Binding Site (SEQ ID NO: 112) GAAGAAGCTGCAGGAGGT Synthetic: Zinc Finger ZF8/7 Binding Site (SEQ ID NO: 113) GCTGGAGGGGAAGTGGTC Synthetic: Zinc Finger ZF6/10 & ZF8/7 Binding Site (SEQ ID NO: 114) GAAGAAGCTGCAGGAGGTGCTGGAGGGGAAGTGGTC Synthetic: Zinc Finger ZF6/10 & ZF8/7 Binding Site 8x Repeat Example (SEQ ID NO: 115) TGAAGAAGCTGCAGGAGGTGCTGGAGGGGAAGTGGTCCGGATCTTGAAGAAGCTGCAGGAGGTGCTGGAGGG GAAGTGGTCCGGATCTTGAAGAAGCTGCAGGAGGTGCTGGAGGGGAAGTGGTCCGGATCTTGAAGAAGCTGC AGGAGGTGCTGGAGGGGAAGTGGTCCGGATCTTGAAGAAGCTGCAGGAGGTGCTGGAGGGGAAGTGGTCCGG ATCTTGAAGAAGCTGCAGGAGGTGCTGGAGGGGAAGTGGTCCGGATCTTGAAGAAGCTGCAGGAGGTGCTGG AGGGGAAGTGGTCCGGATCTTGAAGAAGCTGCAGGAGGTGCTGGAGGGGAAGTGGTCC Synthetic: Zinc Finger ZF9 Binding Site (SEQ ID NO: 116) GTAGATGGA Synthetic: Zinc Finger MK10 Binding Site (SEQ ID NO: 117) CGGCGTAGCCGATGTCGCGC Synthetic: Zinc Finger 268 Binding Site (SEQ ID NO: 118) AAGGGTTCA Synthetic: Zinc Finger NRE Binding Site (SEQ ID NO: 119) GCGTGGGCG Synthetic: Zinc Finger 268/NRE or 268//NRE Binding Site Example 1 (SEQ ID NO: 120) AAGGGTTCAGCGTGGGCG Synthetic: Zinc Finger 268/NRE or 268//NRE Binding Site Example 2 (SEQ ID NO: 121) AAGGGTTCAGGCGTGGGCG Synthetic: Zinc Finger 268/NRE or 268//NRE Binding Site Example 3 (SEQ ID NO: 122) AAGGGTTCAGTGCGTGGGCG Synthetic: Fokl Zinc Finger Nuclease 17-2 & 18-2 Binding Site in GFP (SEQ ID NO: 123) GATCCGCCACAACATCGAGGACGGCA
LITERATURE CITED
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OTHER EMBODIMENTS
[0318] It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.
Sequence CWU
1
1
1231396PRTHomo sapiens 1Met Glu Leu Asp His Arg Thr Ser Gly Gly Leu His
Ala Tyr Pro Gly1 5 10
15Pro Arg Gly Gly Gln Val Ala Lys Pro Asn Val Ile Leu Gln Ile Gly
20 25 30Lys Cys Arg Ala Glu Met Leu
Glu His Val Arg Arg Thr His Arg His 35 40
45Leu Leu Ala Glu Val Ser Lys Gln Val Glu Arg Glu Leu Lys Gly
Leu 50 55 60His Arg Ser Val Gly Lys
Leu Glu Ser Asn Leu Asp Gly Tyr Val Pro65 70
75 80Thr Ser Asp Ser Gln Arg Trp Lys Lys Ser Ile
Lys Ala Cys Leu Cys 85 90
95Arg Cys Gln Glu Thr Ile Ala Asn Leu Glu Arg Trp Val Lys Arg Glu
100 105 110Met His Val Trp Arg Glu
Val Phe Tyr Arg Leu Glu Arg Trp Ala Asp 115 120
125Arg Leu Glu Ser Thr Gly Gly Lys Tyr Pro Val Gly Ser Glu
Ser Ala 130 135 140Arg His Thr Val Ser
Val Gly Val Gly Gly Pro Glu Ser Tyr Cys His145 150
155 160Glu Ala Asp Gly Tyr Asp Tyr Thr Val Ser
Pro Tyr Ala Ile Thr Pro 165 170
175Pro Pro Ala Ala Gly Glu Leu Pro Gly Gln Glu Pro Ala Glu Ala Gln
180 185 190Gln Tyr Gln Pro Trp
Val Pro Gly Glu Asp Gly Gln Pro Ser Pro Gly 195
200 205Val Asp Thr Gln Ile Phe Glu Asp Pro Arg Glu Phe
Leu Ser His Leu 210 215 220Glu Glu Tyr
Leu Arg Gln Val Gly Gly Ser Glu Glu Tyr Trp Leu Ser225
230 235 240Gln Ile Gln Asn His Met Asn
Gly Pro Ala Lys Lys Trp Trp Glu Phe 245
250 255Lys Gln Gly Ser Val Lys Asn Trp Val Glu Phe Lys
Lys Glu Phe Leu 260 265 270Gln
Tyr Ser Glu Gly Thr Leu Ser Arg Glu Ala Ile Gln Arg Glu Leu 275
280 285Asp Leu Pro Gln Lys Gln Gly Glu Pro
Leu Asp Gln Phe Leu Trp Arg 290 295
300Lys Arg Asp Leu Tyr Gln Thr Leu Tyr Val Asp Ala Asp Glu Glu Glu305
310 315 320Ile Ile Gln Tyr
Val Val Gly Thr Leu Gln Pro Lys Leu Lys Arg Phe 325
330 335Leu Arg His Pro Leu Pro Lys Thr Leu Glu
Gln Leu Ile Gln Arg Gly 340 345
350Met Glu Val Gln Asp Asp Leu Glu Gln Ala Ala Glu Pro Ala Gly Pro
355 360 365His Leu Pro Val Glu Asp Glu
Ala Glu Thr Leu Thr Pro Ala Pro Asn 370 375
380Ser Glu Ser Val Ala Ser Asp Arg Thr Gln Pro Glu385
390 3952584PRTHomo sapiens 2Met Ile Phe Ala Gly Lys Ala
Pro Ser Asn Thr Ser Thr Leu Met Lys1 5 10
15Phe Tyr Ser Leu Leu Leu Tyr Ser Leu Leu Phe Ser Phe
Pro Phe Leu 20 25 30Cys His
Pro Leu Pro Leu Pro Ser Tyr Leu His His Thr Ile Asn Leu 35
40 45Thr His Ser Leu Leu Ala Ala Ser Asn Pro
Ser Leu Val Asn Asn Cys 50 55 60Trp
Leu Cys Ile Ser Leu Ser Ser Ser Ala Tyr Thr Ala Val Pro Ala65
70 75 80Val Gln Thr Asp Trp Ala
Thr Ser Pro Ile Ser Leu His Leu Arg Thr 85
90 95Ser Phe Asn Ser Pro His Leu Tyr Pro Pro Glu Glu
Leu Ile Tyr Phe 100 105 110Leu
Asp Arg Ser Ser Lys Thr Ser Pro Asp Ile Ser His Gln Gln Ala 115
120 125Ala Ala Leu Leu Arg Thr Tyr Leu Lys
Asn Leu Ser Pro Tyr Ile Asn 130 135
140Ser Thr Pro Pro Ile Phe Gly Pro Leu Thr Thr Gln Thr Thr Ile Pro145
150 155 160Val Ala Ala Pro
Leu Cys Ile Ser Trp Gln Arg Pro Thr Gly Ile Pro 165
170 175Leu Gly Asn Leu Ser Pro Ser Arg Cys Ser
Phe Thr Leu His Leu Arg 180 185
190Ser Pro Thr Thr Asn Ile Asn Glu Thr Ile Gly Ala Phe Gln Leu His
195 200 205Ile Thr Asp Lys Pro Ser Ile
Asn Thr Asp Lys Leu Lys Asn Ile Ser 210 215
220Ser Asn Tyr Cys Leu Gly Arg His Leu Pro Cys Ile Ser Leu His
Pro225 230 235 240Trp Leu
Ser Ser Pro Cys Ser Ser Asp Ser Pro Pro Arg Pro Ser Ser
245 250 255Cys Leu Leu Ile Pro Ser Pro
Glu Asn Asn Ser Glu Arg Leu Leu Val 260 265
270Asp Thr Arg Arg Phe Leu Ile His His Glu Asn Arg Thr Phe
Pro Ser 275 280 285Thr Gln Leu Pro
His Gln Ser Pro Leu Gln Pro Leu Thr Ala Ala Ala 290
295 300Leu Ala Gly Ser Leu Gly Val Trp Val Gln Asp Thr
Pro Phe Ser Thr305 310 315
320Pro Ser His Leu Phe Thr Leu His Leu Gln Phe Cys Leu Ala Gln Gly
325 330 335Leu Phe Phe Leu Cys
Gly Ser Ser Thr Tyr Met Cys Leu Pro Ala Asn 340
345 350Trp Thr Gly Thr Cys Thr Leu Val Phe Leu Thr Pro
Lys Ile Gln Phe 355 360 365Ala Asn
Gly Thr Glu Glu Leu Pro Val Pro Leu Met Thr Pro Thr Gln 370
375 380Gln Lys Arg Val Ile Pro Leu Ile Pro Leu Met
Val Gly Leu Gly Leu385 390 395
400Ser Ala Ser Thr Val Ala Leu Gly Thr Gly Ile Ala Gly Ile Ser Thr
405 410 415Ser Val Met Thr
Phe Arg Ser Leu Ser Asn Asp Phe Ser Ala Ser Ile 420
425 430Thr Asp Ile Ser Gln Thr Leu Ser Val Leu Gln
Ala Gln Val Asp Ser 435 440 445Leu
Ala Ala Val Val Leu Gln Asn Arg Arg Gly Leu Asp Leu Leu Thr 450
455 460Ala Glu Lys Gly Gly Leu Cys Ile Phe Leu
Asn Glu Glu Cys Cys Phe465 470 475
480Tyr Leu Asn Gln Ser Gly Leu Val Tyr Asp Asn Ile Lys Lys Leu
Lys 485 490 495Asp Arg Ala
Gln Lys Leu Ala Asn Gln Ala Ser Asn Tyr Ala Glu Pro 500
505 510Pro Trp Ala Leu Ser Asn Trp Met Ser Trp
Val Leu Pro Ile Val Ser 515 520
525Pro Leu Ile Pro Ile Phe Leu Leu Leu Leu Phe Gly Pro Cys Ile Phe 530
535 540Arg Leu Val Ser Gln Phe Ile Gln
Asn Arg Ile Gln Ala Ile Thr Asn545 550
555 560His Ser Ile Arg Gln Met Phe Leu Leu Thr Ser Pro
Gln Tyr His Pro 565 570
575Leu Pro Gln Asp Leu Pro Ser Ala 5803563PRTHomo sapiens 3Met
Ile Phe Ala Gly Arg Ala Ser Ser Asn Thr Ser Thr Leu Met Lys1
5 10 15Phe Tyr Ser Leu Leu Leu Tyr
Ser Leu Leu Phe Ser Phe Pro Ile Leu 20 25
30Cys His Pro Leu Pro Leu Pro Ser Tyr Leu His His Thr Ile
Asn Leu 35 40 45Thr His Ser Leu
Leu Ala Val Ser Asn Pro Ser Leu Ala Lys Asn Cys 50 55
60Trp Leu Cys Ile Ser Leu Pro Ser Ser Ala Tyr Pro Ala
Val Pro Ala65 70 75
80Leu Gln Thr Asp Trp Gly Thr Ser Pro Val Ser Pro His Leu Arg Thr
85 90 95Ser Phe Asn Ser Pro His
Leu Tyr Pro Pro Glu Lys Leu Ile Tyr Phe 100
105 110Leu Asp Arg Ser Ser Lys Thr Ser Pro Asp Ile Ser
His Gln Gln Ala 115 120 125Ala Ala
Leu Leu Cys Thr Tyr Leu Lys Asn Leu Ser Pro Tyr Ile Asn 130
135 140Ser Thr Pro Pro Thr Phe Gly Pro Leu Thr Thr
Gln Thr Thr Ile Pro145 150 155
160Val Ala Ala Pro Leu Cys Ile Ser Arg Gln Arg Pro Thr Gly Ile Pro
165 170 175Leu Gly Asn Leu
Ser Pro Ser Arg Cys Ser Phe Thr Leu His Leu Arg 180
185 190Ser Pro Thr Thr His Ile Thr Glu Thr Asn Gly
Ala Phe Gln Leu His 195 200 205Ile
Thr Asp Lys Pro Ser Ile Asn Thr Asp Lys Leu Lys Asn Val Ser 210
215 220Ser Asn Tyr Cys Leu Gly Arg His Leu Ser
Cys Ile Ser Leu His Pro225 230 235
240Trp Leu Phe Ser Pro Cys Ser Ser Asp Ser Pro Pro Arg Pro Ser
Ser 245 250 255Cys Leu Leu
Ile Pro Ser Pro Lys Asn Asn Ser Glu Ser Leu Leu Val 260
265 270Asp Ala Gln Arg Phe Leu Ile Tyr His Glu
Asn Arg Thr Ser Pro Ser 275 280
285Thr Gln Leu Pro His Gln Ser Pro Leu Gln Pro Leu Thr Ala Ala Pro 290
295 300Leu Gly Gly Ser Leu Arg Val Trp
Val Gln Asp Thr Pro Phe Ser Thr305 310
315 320Pro Ser His Leu Phe Thr Leu His Leu Gln Phe Cys
Leu Val Gln Ser 325 330
335Leu Phe Phe Leu Cys Gly Ser Ser Thr Tyr Met Cys Leu Pro Ala Asn
340 345 350Trp Thr Gly Thr Cys Thr
Leu Val Phe Leu Thr Ser Lys Ile Gln Phe 355 360
365Ala Asn Gly Thr Glu Glu Leu Pro Val Pro Leu Met Thr Pro
Thr Arg 370 375 380Gln Lys Arg Val Ile
Pro Leu Ile Pro Leu Met Val Gly Leu Gly Leu385 390
395 400Ser Ala Ser Thr Val Ala Leu Gly Thr Gly
Ile Ala Gly Ile Ser Thr 405 410
415Ser Val Thr Thr Phe Arg Ile Leu Ser Asn Asp Phe Ser Ala Ser Ile
420 425 430Thr Asp Ile Ser Gln
Thr Leu Ser Gly Leu Gln Ala Gln Val Asp Ser 435
440 445Ser Ala Ala Val Val Leu Gln Asn Arg Gln Gly Leu
Asp Leu Leu Thr 450 455 460Ala Glu Lys
Gly Gly Leu Cys Ile Phe Leu Asn Glu Glu Ser Tyr Phe465
470 475 480Tyr Leu Asn Gln Ser Gly Leu
Val Tyr Asp Asn Ile Lys Lys Leu Lys 485
490 495Asp Lys Ala Gln Asn Leu Ala Asn Gln Ala Ser Asn
Tyr Ala Glu Pro 500 505 510Pro
Trp Pro Leu Ser Asn Trp Met Ser Trp Val Leu Pro Ile Leu Ser 515
520 525Pro Leu Ile Pro Ile Phe Leu Leu Leu
Phe Phe Arg Pro Cys Ile Phe 530 535
540His Leu Val Ser Gln Phe Ile Gln Asn His Ile Gln Ala Ile Thr Asp545
550 555 560His Ser
Ile4555PRTHomo sapiens 4Met Ile Leu Ala Gly Arg Ala Pro Ser Asn Thr Ser
Thr Leu Met Lys1 5 10
15Phe Tyr Ser Leu Leu Leu Tyr Ser Leu Leu Phe Ser Phe Pro Phe Leu
20 25 30Tyr His Pro Leu Pro Leu Pro
Ser Tyr Leu His His Thr Ile Asn Leu 35 40
45Thr His Ser Leu Pro Ala Ala Ser Asn Pro Ser Leu Ala Asn Asn
Cys 50 55 60Trp Leu Cys Ile Ser Leu
Ser Ser Ser Ala Tyr Ile Ala Val Pro Thr65 70
75 80Leu Gln Thr Asp Arg Ala Thr Ser Pro Val Ser
Leu His Leu Arg Thr 85 90
95Ser Phe Asn Ser Pro His Leu Tyr Pro Pro Glu Glu Leu Ile Tyr Phe
100 105 110Leu Asp Arg Ser Ser Lys
Thr Ser Pro Asp Ile Ser His Gln Pro Ala 115 120
125Ala Ala Leu Leu His Ile Tyr Leu Lys Asn Leu Ser Pro Tyr
Ile Asn 130 135 140Ser Thr Pro Pro Ile
Phe Gly Pro Leu Thr Thr Gln Thr Thr Ile Pro145 150
155 160Val Ala Ala Pro Leu Cys Ile Ser Arg Gln
Arg Pro Thr Gly Ile Pro 165 170
175Leu Gly Asn Ile Ser Pro Ser Arg Cys Ser Phe Thr Leu His Leu Gln
180 185 190Ser Pro Thr Thr His
Val Thr Glu Thr Ile Gly Val Phe Gln Leu His 195
200 205Ile Ile Asp Lys Pro Ser Ile Asn Thr Asp Lys Leu
Lys Asn Val Ser 210 215 220Ser Asn Tyr
Cys Leu Gly Arg His Leu Pro Tyr Ile Ser Leu His Pro225
230 235 240Trp Leu Pro Ser Pro Cys Ser
Ser Asp Ser Pro Pro Arg Pro Ser Ser 245
250 255Cys Leu Leu Thr Pro Ser Pro Gln Asn Asn Ser Glu
Arg Leu Leu Val 260 265 270Asp
Thr Gln Arg Phe Leu Ile His His Glu Asn Arg Thr Ser Ser Ser 275
280 285Met Gln Leu Ala His Gln Ser Pro Leu
Gln Pro Leu Thr Ala Ala Ala 290 295
300Leu Ala Gly Ser Leu Gly Val Trp Val Gln Asp Thr Pro Phe Ser Thr305
310 315 320Pro Ser His Pro
Phe Ser Leu His Leu Gln Phe Cys Leu Thr Gln Gly 325
330 335Leu Phe Phe Leu Cys Gly Ser Ser Thr Tyr
Met Cys Leu Pro Ala Asn 340 345
350Trp Thr Gly Thr Cys Thr Leu Val Phe Leu Thr Pro Lys Ile Gln Phe
355 360 365Ala Asn Gly Thr Lys Glu Leu
Pro Val Pro Leu Met Thr Leu Thr Pro 370 375
380Gln Lys Arg Val Ile Pro Leu Ile Pro Leu Met Val Gly Leu Gly
Leu385 390 395 400Ser Ala
Ser Thr Ile Ala Leu Ser Thr Gly Ile Ala Gly Ile Ser Thr
405 410 415Ser Val Thr Thr Phe Arg Ser
Pro Ser Asn Asp Phe Ser Ala Ser Ile 420 425
430Thr Asp Ile Ser Gln Thr Leu Ser Val Leu Gln Ala Gln Val
Asp Ser 435 440 445Leu Ala Ala Val
Val Leu Gln Asn Arg Arg Gly Leu Gly Leu Ser Ile 450
455 460Leu Leu Asn Glu Glu Cys Cys Phe Tyr Leu Asn Gln
Ser Gly Leu Val465 470 475
480Tyr Glu Asn Ile Lys Lys Leu Lys Asp Arg Ala Gln Lys Leu Ala Asn
485 490 495Gln Ala Ser Asn Tyr
Ala Glu Ser Pro Trp Ala Leu Ser Asn Trp Met 500
505 510Ser Trp Val Leu Pro Ile Leu Ser Pro Leu Ile Pro
Ile Phe Leu Leu 515 520 525Leu Leu
Phe Gly Pro Cys Ile Phe His Leu Val Ser Gln Phe Ile Gln 530
535 540Asn Arg Ile Gln Ala Ile Thr Asn His Ser
Ile545 550 5555698PRTHomo sapiens 5Met
His Pro Ser Glu Met Gln Arg Lys Ala Pro Pro Arg Arg Arg Arg1
5 10 15His Arg Asn Arg Ala Pro Leu
Thr His Lys Met Asn Lys Met Val Thr 20 25
30Ser Glu Gln Met Lys Leu Pro Ser Thr Lys Lys Ala Glu Pro
Pro Thr 35 40 45Trp Ala Gln Leu
Lys Lys Leu Thr Gln Leu Ala Thr Lys Tyr Leu Glu 50 55
60Asn Thr Lys Val Thr Gln Thr Pro Glu Ser Met Leu Leu
Ala Ala Leu65 70 75
80Met Ile Val Ser Met Val Val Ser Leu Pro Met Pro Ala Gly Ala Ala
85 90 95Ala Ala Asn Tyr Thr Asn
Trp Ala Tyr Val Pro Phe Pro Pro Leu Ile 100
105 110Arg Ala Val Thr Trp Met Asp Asn Pro Ile Glu Val
Tyr Val Asn Asp 115 120 125Ser Val
Trp Val His Gly Pro Ile Asp Asp Arg Cys Pro Ala Lys Pro 130
135 140Glu Glu Glu Gly Met Met Ile Asn Ile Ser Ile
Gly Tyr His Tyr Pro145 150 155
160Pro Ile Cys Leu Gly Arg Ala Pro Gly Cys Leu Met Pro Ala Val Gln
165 170 175Asn Trp Leu Val
Glu Val Pro Thr Val Ser Pro Ile Ser Arg Phe Thr 180
185 190Tyr Asn Met Val Ser Gly Met Ser Leu Arg Pro
Arg Val Asn Tyr Leu 195 200 205Gln
Asp Phe Ser Tyr Gln Arg Ser Leu Lys Phe Arg Pro Lys Gly Lys 210
215 220Pro Cys Pro Lys Glu Ile Pro Lys Glu Ser
Lys Asn Thr Glu Val Leu225 230 235
240Val Trp Glu Glu Cys Val Ala Asn Ser Val Val Ile Leu Gln Asn
Asn 245 250 255Glu Phe Gly
Thr Ile Ile Asp Trp Ala Pro Arg Gly Gln Phe Tyr His 260
265 270Asn Cys Ser Gly Gln Thr Gln Ser Cys Pro
Ser Ala Gln Val Ser Pro 275 280
285Ala Val Asp Ser Asp Leu Thr Glu Ser Leu Asp Lys His Lys His Lys 290
295 300Lys Leu Gln Ser Phe Tyr Pro Trp
Glu Trp Gly Glu Lys Gly Ile Ser305 310
315 320Thr Pro Arg Pro Lys Ile Ile Ser Pro Val Ser Gly
Pro Glu His Pro 325 330
335Glu Leu Trp Arg Leu Thr Val Ala Ser His His Ile Arg Ile Trp Ser
340 345 350Gly Asn Gln Thr Leu Glu
Thr Arg Asp Arg Lys Pro Phe Tyr Thr Val 355 360
365Asp Leu Asn Ser Ser Leu Thr Val Pro Leu Gln Ser Cys Val
Lys Pro 370 375 380Pro Tyr Met Leu Val
Val Gly Asn Ile Val Ile Lys Pro Asp Ser Gln385 390
395 400Thr Ile Thr Cys Glu Asn Cys Arg Leu Leu
Thr Cys Ile Asp Ser Thr 405 410
415Phe Asn Trp Gln His Arg Ile Leu Leu Val Arg Ala Arg Glu Gly Val
420 425 430Trp Ile Pro Val Ser
Met Asp Arg Pro Trp Glu Ala Ser Pro Ser Ile 435
440 445His Ile Leu Thr Glu Val Leu Lys Gly Val Leu Asn
Arg Ser Lys Arg 450 455 460Phe Ile Phe
Thr Leu Ile Ala Val Ile Met Gly Leu Ile Ala Val Thr465
470 475 480Ala Met Ala Ala Val Ala Gly
Val Ala Leu His Ser Phe Val Gln Ser 485
490 495Val Asn Phe Val Asn Asp Trp Gln Lys Asn Ser Thr
Arg Leu Trp Asn 500 505 510Ser
Gln Ser Ser Ile Asp Gln Lys Leu Ala Asn Gln Ile Asn Asp Leu 515
520 525Arg Gln Thr Val Ile Trp Met Gly Asp
Arg Leu Met Ser Leu Glu His 530 535
540Arg Phe Gln Leu Gln Cys Asp Trp Asn Thr Ser Asp Phe Cys Ile Thr545
550 555 560Pro Gln Ile Tyr
Asn Glu Ser Glu His His Trp Asp Met Val Arg Arg 565
570 575His Leu Gln Gly Arg Glu Asp Asn Leu Thr
Leu Asp Ile Ser Lys Leu 580 585
590Lys Glu Gln Ile Phe Glu Ala Ser Lys Ala His Leu Asn Leu Val Pro
595 600 605Gly Thr Glu Ala Ile Ala Gly
Val Ala Asp Gly Leu Ala Asn Leu Asn 610 615
620Pro Val Thr Trp Val Lys Thr Ile Gly Ser Thr Thr Ile Ile Asn
Leu625 630 635 640Ile Leu
Ile Leu Val Cys Leu Phe Cys Leu Leu Leu Val Cys Arg Phe
645 650 655Thr Gln Gln Leu Arg Arg Asp
Ser Tyr His Arg Glu Arg Ala Met Met 660 665
670Thr Met Val Val Leu Ser Lys Arg Lys Gly Gly Asn Val Gly
Lys Ser 675 680 685Lys Arg Asp Gln
Ile Val Thr Val Ser Val 690 6956699PRTHomo sapiens
6Met Asn Pro Ser Glu Met Gln Arg Lys Ala Pro Pro Arg Arg Arg Arg1
5 10 15His Arg Asn Arg Ala Pro
Leu Thr His Lys Met Asn Lys Met Val Thr 20 25
30Ser Glu Glu Gln Met Lys Leu Pro Ser Thr Lys Lys Ala
Glu Pro Pro 35 40 45Thr Trp Ala
Gln Leu Lys Lys Leu Thr Gln Leu Ala Thr Lys Tyr Leu 50
55 60Glu Asn Thr Lys Val Thr Gln Thr Pro Glu Ser Met
Leu Leu Ala Ala65 70 75
80Leu Met Ile Val Ser Met Val Val Ser Leu Pro Met Pro Ala Gly Ala
85 90 95Ala Ala Ala Asn Tyr Thr
Tyr Trp Ala Tyr Val Pro Phe Pro Pro Leu 100
105 110Ile Arg Ala Val Thr Trp Met Asp Asn Pro Thr Glu
Val Tyr Val Asn 115 120 125Asp Ser
Val Trp Val Pro Gly Pro Ile Asp Asp Arg Cys Pro Ala Lys 130
135 140Pro Glu Glu Glu Gly Met Met Ile Asn Ile Ser
Ile Gly Tyr His Tyr145 150 155
160Pro Pro Ile Cys Leu Gly Arg Ala Pro Gly Cys Leu Met Pro Ala Val
165 170 175Gln Asn Trp Leu
Val Glu Val Pro Thr Val Ser Pro Ile Cys Arg Phe 180
185 190Thr Tyr His Met Val Ser Gly Met Ser Leu Arg
Pro Arg Val Asn Tyr 195 200 205Leu
Gln Asp Phe Ser Tyr Gln Arg Ser Leu Lys Phe Arg Pro Lys Gly 210
215 220Lys Pro Cys Pro Lys Glu Ile Pro Lys Glu
Ser Lys Asn Thr Glu Val225 230 235
240Leu Val Trp Glu Glu Cys Val Ala Asn Ser Ala Val Ile Leu Gln
Asn 245 250 255Asn Glu Phe
Gly Thr Ile Ile Asp Trp Ala Pro Arg Gly Gln Phe Tyr 260
265 270His Asn Cys Ser Gly Gln Thr Gln Ser Cys
Pro Ser Ala Gln Val Ser 275 280
285Pro Ala Val Asp Ser Asp Leu Thr Glu Ser Leu Asp Lys His Lys His 290
295 300Lys Lys Leu Gln Ser Phe Tyr Pro
Trp Glu Trp Gly Glu Lys Gly Ile305 310
315 320Ser Thr Pro Arg Pro Lys Ile Val Ser Pro Val Ser
Gly Pro Glu His 325 330
335Pro Glu Leu Trp Arg Leu Thr Val Ala Ser His His Ile Arg Ile Trp
340 345 350Ser Gly Asn Gln Thr Leu
Glu Thr Arg Asp Arg Lys Pro Phe Tyr Thr 355 360
365Ile Asp Leu Asn Ser Ser Leu Thr Val Pro Leu Gln Ser Cys
Val Lys 370 375 380Pro Pro Tyr Met Leu
Val Val Gly Asn Ile Val Ile Lys Pro Asp Ser385 390
395 400Gln Thr Ile Thr Cys Glu Asn Cys Arg Leu
Leu Thr Cys Ile Asp Ser 405 410
415Thr Phe Asn Trp Gln His Arg Ile Leu Leu Val Arg Ala Arg Glu Gly
420 425 430Val Trp Ile Pro Val
Ser Met Asp Arg Pro Trp Glu Ala Ser Pro Ser 435
440 445Val His Ile Leu Thr Glu Val Leu Lys Gly Val Leu
Asn Arg Ser Lys 450 455 460Arg Phe Ile
Phe Thr Leu Ile Ala Val Ile Met Gly Leu Ile Ala Val465
470 475 480Thr Ala Thr Ala Ala Val Ala
Gly Val Ala Leu His Ser Ser Val Gln 485
490 495Ser Val Asn Phe Val Asn Asp Trp Gln Lys Asn Ser
Thr Arg Leu Trp 500 505 510Asn
Ser Gln Ser Ser Ile Asp Gln Lys Leu Ala Asn Gln Ile Asn Asp 515
520 525Leu Arg Gln Thr Val Ile Trp Met Gly
Asp Arg Leu Met Ser Leu Glu 530 535
540His Arg Phe Gln Leu Gln Cys Asp Trp Asn Thr Ser Asp Phe Cys Ile545
550 555 560Thr Pro Gln Ile
Tyr Asn Glu Ser Glu His His Trp Asp Met Val Arg 565
570 575Arg His Leu Gln Gly Arg Glu Asp Asn Leu
Thr Leu Asp Ile Ser Lys 580 585
590Leu Lys Glu Gln Ile Phe Glu Ala Ser Lys Ala His Leu Asn Leu Val
595 600 605Pro Gly Thr Glu Ala Ile Ala
Gly Val Ala Asp Gly Leu Ala Asn Leu 610 615
620Asn Pro Val Thr Trp Val Lys Thr Ile Gly Ser Thr Thr Ile Ile
Asn625 630 635 640Leu Ile
Leu Ile Leu Val Cys Leu Phe Cys Leu Leu Leu Val Cys Arg
645 650 655Cys Thr Gln Gln Leu Arg Arg
Asp Ser Asp His Arg Glu Arg Ala Met 660 665
670Met Thr Met Ala Val Leu Ser Lys Arg Lys Gly Gly Asn Val
Gly Lys 675 680 685Ser Lys Arg Asp
Gln Ile Val Thr Val Ser Val 690 6957699PRTHomo sapiens
7Met Asn Pro Ser Glu Met Gln Arg Lys Ala Pro Pro Arg Arg Arg Arg1
5 10 15His Arg Asn Arg Ala Pro
Leu Thr His Lys Met Asn Lys Met Val Thr 20 25
30Ser Glu Glu Gln Met Lys Leu Pro Ser Thr Lys Lys Ala
Glu Pro Pro 35 40 45Thr Trp Ala
Gln Leu Lys Lys Leu Thr Gln Leu Ala Thr Lys Tyr Leu 50
55 60Glu Asn Thr Lys Val Thr Gln Thr Pro Glu Ser Met
Leu Leu Ala Ala65 70 75
80Leu Met Ile Val Ser Met Val Val Ser Leu Pro Met Pro Ala Gly Ala
85 90 95Ala Ala Ala Asn Tyr Thr
Tyr Trp Ala Tyr Val Pro Phe Pro Pro Leu 100
105 110Ile Arg Ala Val Thr Trp Met Asp Asn Pro Ile Glu
Val Tyr Val Asn 115 120 125Asp Ser
Val Trp Val Pro Gly Pro Thr Asp Asp His Cys Pro Ala Lys 130
135 140Pro Glu Glu Glu Gly Met Met Ile Asn Ile Ser
Ile Gly Tyr Arg Tyr145 150 155
160Pro Pro Ile Cys Leu Gly Arg Ala Pro Gly Cys Leu Met Pro Ala Val
165 170 175Gln Asn Trp Leu
Val Glu Val Pro Thr Val Ser Pro Ile Ser Arg Phe 180
185 190Thr Tyr His Met Val Ser Gly Met Ser Leu Arg
Pro Arg Val Asn Tyr 195 200 205Leu
Gln Asp Phe Ser Tyr Gln Arg Ser Phe Lys Phe Arg Pro Lys Gly 210
215 220Lys Pro Cys Pro Lys Glu Ile Pro Lys Glu
Ser Lys Asn Thr Glu Val225 230 235
240Leu Val Trp Glu Glu Cys Val Ala Asn Ser Ala Val Ile Leu Gln
Asn 245 250 255Asn Glu Phe
Gly Thr Ile Ile Asp Trp Ala Pro Arg Gly Gln Phe Tyr 260
265 270His Asn Cys Ser Gly Gln Thr Gln Ser Cys
Pro Ser Ala Gln Val Ser 275 280
285Pro Ala Val Asp Ser Asp Leu Thr Glu Ser Leu Asp Lys His Lys His 290
295 300Lys Lys Leu Gln Ser Phe Tyr Pro
Trp Glu Trp Gly Glu Lys Gly Ile305 310
315 320Ser Thr Pro Arg Pro Lys Ile Ile Ser Pro Val Ser
Gly Pro Glu His 325 330
335Pro Glu Leu Trp Arg Leu Thr Val Ala Ser His His Ile Arg Ile Trp
340 345 350Ser Gly Asn Gln Thr Leu
Glu Thr Arg Asp Arg Lys Pro Phe Tyr Thr 355 360
365Val Asp Leu Asn Ser Ser Val Thr Val Pro Leu Gln Ser Cys
Ile Lys 370 375 380Pro Pro Tyr Met Leu
Val Val Gly Asn Ile Val Ile Lys Pro Asp Ser385 390
395 400Gln Thr Ile Thr Cys Glu Asn Cys Arg Leu
Leu Thr Cys Ile Asp Ser 405 410
415Thr Phe Asn Trp Gln His Arg Ile Leu Leu Val Arg Ala Arg Glu Gly
420 425 430Val Trp Ile Pro Val
Ser Met Asp Arg Pro Trp Glu Thr Ser Pro Ser 435
440 445Ile His Thr Leu Thr Glu Val Leu Lys Gly Val Leu
Asn Arg Ser Lys 450 455 460Arg Phe Ile
Phe Thr Leu Ile Ala Val Ile Met Gly Leu Ile Ala Val465
470 475 480Thr Ala Thr Ala Ala Val Ala
Gly Val Ala Leu His Ser Ser Val Gln 485
490 495Ser Val Asn Phe Val Asn Asp Trp Gln Lys Asn Ser
Thr Arg Leu Trp 500 505 510Asn
Ser Gln Ser Ser Ile Asp Gln Lys Leu Ala Asn Gln Ile Asn Asp 515
520 525Leu Arg Gln Thr Val Ile Trp Met Gly
Asp Arg Leu Met Ser Leu Glu 530 535
540His Arg Phe Gln Leu Gln Cys Asp Trp Asn Thr Ser Asp Phe Ser Ile545
550 555 560Thr Pro Gln Ile
Tyr Asn Glu Ser Glu His His Trp Asp Met Val Arg 565
570 575Arg His Leu Gln Gly Arg Glu Asp Asn Leu
Thr Leu Asp Ile Ser Lys 580 585
590Leu Lys Glu Gln Ile Phe Glu Ala Ser Lys Ala His Leu Asn Leu Val
595 600 605Pro Gly Thr Glu Ala Ile Ala
Gly Val Ala Asp Gly Leu Ala Asn Leu 610 615
620Asn Pro Val Thr Trp Val Lys Thr Ile Gly Ser Thr Thr Ile Ile
Asn625 630 635 640Leu Ile
Leu Ile Leu Val Cys Leu Phe Cys Leu Leu Leu Val Cys Arg
645 650 655Cys Thr Gln Gln Leu Arg Arg
Asp Ser Asp His Arg Glu Arg Ala Met 660 665
670Met Thr Met Ala Val Leu Ser Lys Arg Lys Gly Gly Asn Val
Gly Lys 675 680 685Ser Lys Arg Asp
Gln Ile Val Thr Val Ser Val 690 6958698PRTHomo sapiens
8Met Asn Pro Ser Glu Met Gln Arg Lys Ala Pro Pro Arg Arg Arg Arg1
5 10 15His Arg Asn Arg Ala Pro
Leu Thr His Lys Met Asn Lys Met Val Thr 20 25
30Ser Glu Glu Gln Met Lys Leu Pro Ser Thr Lys Lys Ala
Glu Pro Pro 35 40 45Thr Trp Ala
Gln Leu Lys Lys Leu Thr Gln Leu Ala Thr Lys Tyr Leu 50
55 60Glu Asn Thr Lys Val Thr Gln Thr Pro Glu Ser Met
Leu Leu Ala Ala65 70 75
80Leu Met Ile Val Ser Met Val Val Ser Leu Pro Met Pro Ala Gly Ala
85 90 95Ala Ala Ala Asn Tyr Thr
Asn Trp Ala Tyr Val Pro Phe Pro Pro Leu 100
105 110Ile Arg Ala Val Thr Trp Met Asp Asn Pro Ile Glu
Val Tyr Val Asn 115 120 125Asp Ser
Val Trp Val Pro Gly Pro Ile Asp Asp Arg Cys Pro Ala Lys 130
135 140Pro Glu Glu Glu Gly Met Met Ile Asn Ile Ser
Ile Gly Tyr Arg Tyr145 150 155
160Pro Ile Cys Leu Gly Arg Ala Pro Gly Cys Leu Met Pro Ala Val Gln
165 170 175Asn Trp Leu Val
Glu Val Pro Ile Val Ser Pro Ile Cys Arg Phe Thr 180
185 190Tyr His Met Val Ser Gly Met Ser Leu Arg Pro
Arg Val Asn Tyr Leu 195 200 205Gln
Asp Phe Ser Tyr Gln Arg Ser Leu Lys Phe Arg Pro Lys Gly Lys 210
215 220Pro Cys Pro Lys Glu Ile Pro Lys Glu Ser
Lys Asn Thr Glu Val Leu225 230 235
240Val Trp Glu Glu Cys Val Ala Asn Ser Ala Val Ile Leu Gln Asn
Asn 245 250 255Glu Phe Gly
Thr Ile Ile Asp Trp Thr Pro Gln Gly Gln Phe Tyr His 260
265 270Asn Cys Ser Gly Gln Thr Gln Ser Cys Pro
Ser Ala Gln Val Ser Pro 275 280
285Ala Val Asp Ser Asp Leu Thr Glu Ser Leu Asp Lys His Lys His Lys 290
295 300Lys Leu Gln Ser Phe Tyr Pro Trp
Glu Trp Gly Glu Lys Gly Ile Ser305 310
315 320Thr Pro Arg Pro Lys Ile Ile Ser Pro Val Ser Gly
Pro Glu His Pro 325 330
335Glu Leu Trp Arg Leu Thr Val Ala Ser His His Ile Arg Ile Trp Ser
340 345 350Gly Asn Gln Thr Leu Glu
Thr Arg Asp Arg Lys Pro Phe Tyr Thr Val 355 360
365Asp Leu Asn Ser Ser Leu Thr Leu Pro Leu Gln Ser Cys Val
Lys Pro 370 375 380Pro Tyr Met Leu Val
Val Gly Asn Ile Val Ile Lys Pro Asp Ser Gln385 390
395 400Thr Ile Thr Cys Glu Asn Cys Arg Leu Leu
Thr Cys Ile Asp Ser Thr 405 410
415Phe Asn Trp Gln His Arg Ile Leu Leu Val Arg Ala Arg Glu Gly Val
420 425 430Trp Ile Pro Val Ser
Met Asp Arg Pro Trp Glu Ala Ser Pro Ser Ile 435
440 445His Ile Leu Thr Glu Val Leu Lys Gly Val Leu Asn
Arg Ser Lys Arg 450 455 460Phe Ile Phe
Thr Leu Ile Ala Val Ile Met Gly Leu Ile Ala Val Thr465
470 475 480Ala Thr Ala Ala Val Ala Gly
Val Ala Leu His Ser Ser Val Gln Ser 485
490 495Val Asn Phe Val Asn Asp Gly Gln Lys Asn Ser Thr
Arg Leu Trp Asn 500 505 510Ser
Gln Ser Ser Ile Asp Gln Lys Leu Ala Asn Gln Ile Asn Asp Leu 515
520 525Arg Gln Thr Val Ile Trp Met Gly Asp
Arg Leu Met Ser Leu Glu His 530 535
540Arg Phe Gln Leu Gln Cys Asp Trp Asn Thr Ser Asp Phe Cys Ile Thr545
550 555 560Pro Gln Ile Tyr
Asn Glu Ser Glu His His Trp Asp Met Val Arg Arg 565
570 575His Leu Gln Gly Arg Glu Asp Asn Leu Thr
Leu Asp Ile Ser Lys Leu 580 585
590Lys Glu Gln Ile Phe Glu Ala Ser Lys Ala His Leu Asn Leu Val Pro
595 600 605Gly Thr Glu Ala Ile Ala Gly
Val Ala Asp Gly Leu Ala Asn Leu Asn 610 615
620Pro Val Thr Trp Val Lys Thr Ile Gly Ser Thr Thr Ile Ile Asn
Leu625 630 635 640Ile Leu
Ile Leu Val Cys Leu Phe Cys Leu Leu Leu Val Cys Arg Cys
645 650 655Thr Gln Gln Leu Arg Arg Asp
Ser Asp His Arg Glu Arg Ala Met Met 660 665
670Thr Met Ala Val Leu Ser Lys Arg Lys Gly Gly Asn Val Gly
Lys Ser 675 680 685Lys Arg Asp Gln
Ile Val Thr Val Ser Val 690 6959699PRTHomo sapiens
9Met Asn Pro Ser Glu Met Gln Arg Lys Ala Pro Pro Arg Arg Arg Arg1
5 10 15His Arg Asn Arg Ala Pro
Leu Thr His Lys Met Asn Lys Met Val Thr 20 25
30Ser Glu Glu Gln Met Lys Leu Pro Ser Thr Lys Lys Ala
Glu Pro Pro 35 40 45Thr Trp Ala
Gln Leu Lys Lys Leu Thr Gln Leu Ala Thr Lys Tyr Leu 50
55 60Glu Asn Thr Lys Val Thr Gln Thr Pro Glu Ser Met
Leu Leu Ala Ala65 70 75
80Leu Met Ile Val Ser Met Val Val Ser Leu Pro Met Pro Ala Gly Ala
85 90 95Ala Ala Ala Asn Tyr Thr
Tyr Trp Ala Tyr Val Pro Phe Pro Pro Leu 100
105 110Ile Arg Ala Val Thr Trp Met Asp Asn Pro Ile Glu
Ile Tyr Val Asn 115 120 125Asp Ser
Val Trp Val Pro Gly Pro Thr Asp Asp Cys Cys Pro Ala Lys 130
135 140Pro Glu Glu Glu Gly Met Met Ile Asn Ile Ser
Ile Gly Tyr Arg Tyr145 150 155
160Pro Pro Ile Cys Leu Gly Arg Ala Pro Gly Cys Leu Met Pro Ala Val
165 170 175Gln Asn Trp Leu
Val Glu Val Pro Thr Val Ser Pro Ile Ser Arg Phe 180
185 190Thr Tyr His Met Val Ser Gly Met Ser Leu Arg
Pro Arg Val Asn Tyr 195 200 205Leu
Gln Asp Phe Ser Tyr Gln Arg Ser Leu Lys Phe Arg Pro Lys Gly 210
215 220Lys Pro Cys Pro Lys Glu Ile Pro Lys Glu
Ser Lys Asn Thr Glu Val225 230 235
240Leu Val Trp Glu Glu Cys Val Ala Asn Ser Ala Val Ile Leu Gln
Asn 245 250 255Asn Glu Phe
Gly Thr Leu Ile Asp Trp Ala Pro Arg Gly Gln Phe Tyr 260
265 270His Asn Cys Ser Gly Gln Thr Gln Ser Cys
Pro Ser Ala Gln Val Ser 275 280
285Pro Ala Val Asp Ser Asp Leu Thr Glu Ser Leu Asp Lys His Lys His 290
295 300Lys Lys Leu Gln Ser Phe Tyr Pro
Trp Glu Trp Gly Glu Lys Gly Ile305 310
315 320Ser Thr Ala Arg Pro Lys Ile Ile Ser Pro Val Ser
Gly Pro Glu His 325 330
335Pro Glu Leu Trp Arg Leu Thr Val Ala Ser His His Ile Arg Ile Trp
340 345 350Ser Gly Asn Gln Thr Leu
Glu Thr Arg Asp Arg Lys Pro Phe Tyr Thr 355 360
365Ile Asp Leu Asn Ser Ser Leu Thr Val Pro Leu Gln Ser Cys
Val Lys 370 375 380Pro Pro Tyr Met Leu
Val Val Gly Asn Ile Val Ile Lys Pro Asp Ser385 390
395 400Gln Thr Ile Thr Cys Glu Asn Cys Arg Leu
Leu Thr Cys Ile Asp Ser 405 410
415Thr Phe Asn Trp Gln His Arg Ile Leu Leu Val Arg Ala Arg Glu Gly
420 425 430Val Trp Ile Pro Val
Ser Met Asp Arg Pro Trp Glu Ala Ser Pro Ser 435
440 445Val His Ile Leu Thr Glu Val Leu Lys Gly Val Leu
Asn Arg Ser Lys 450 455 460Arg Phe Ile
Phe Thr Leu Ile Ala Val Ile Met Gly Leu Ile Ala Val465
470 475 480Thr Ala Thr Ala Ala Val Ala
Gly Val Ala Leu His Ser Ser Val Gln 485
490 495Ser Val Asn Phe Val Asn Asp Trp Gln Asn Asn Ser
Thr Arg Leu Trp 500 505 510Asn
Ser Gln Ser Ser Ile Asp Gln Lys Leu Ala Asn Gln Ile Asn Asp 515
520 525Leu Arg Gln Thr Val Ile Trp Met Gly
Asp Arg Leu Met Ser Leu Glu 530 535
540His Arg Phe Gln Leu Gln Cys Asp Trp Asn Thr Ser Asp Phe Cys Ile545
550 555 560Thr Pro Gln Ile
Tyr Asn Glu Ser Glu His His Trp Asp Met Val Arg 565
570 575Cys His Leu Gln Gly Arg Glu Asp Asn Leu
Thr Leu Asp Ile Ser Lys 580 585
590Leu Lys Glu Gln Ile Phe Glu Ala Ser Lys Ala His Leu Asn Leu Val
595 600 605Pro Gly Thr Glu Ala Ile Ala
Gly Val Ala Asp Gly Leu Ala Asn Leu 610 615
620Asn Thr Val Thr Trp Val Lys Thr Ile Gly Ser Thr Thr Ile Ile
Asn625 630 635 640Leu Ile
Leu Ile Leu Val Cys Leu Phe Cys Leu Leu Leu Val Tyr Arg
645 650 655Cys Thr Gln Gln Leu Arg Arg
Asp Ser Asp His Arg Glu Arg Ala Met 660 665
670Met Thr Met Val Val Leu Ser Lys Arg Lys Gly Gly Asn Val
Gly Lys 675 680 685Ser Lys Arg Asp
Gln Ile Val Thr Val Ser Val 690 69510699PRTHomo
sapiens 10Met Asn Pro Ser Glu Met Gln Arg Lys Ala Pro Pro Arg Arg Arg
Arg1 5 10 15His Arg Asn
Arg Ala Pro Leu Thr His Lys Met Asn Lys Met Val Thr 20
25 30Ser Glu Glu Gln Met Lys Leu Pro Ser Thr
Lys Lys Ala Glu Pro Pro 35 40
45Thr Trp Ala Gln Leu Lys Lys Leu Thr Gln Leu Ala Thr Lys Tyr Leu 50
55 60Glu Asn Thr Lys Val Thr Gln Thr Pro
Glu Ser Met Leu Leu Ala Ala65 70 75
80Leu Met Ile Val Ser Met Val Val Ser Leu Pro Met Pro Ala
Gly Ala 85 90 95Ala Val
Ala Asn Tyr Thr Asn Trp Ala Tyr Val Pro Phe Pro Pro Leu 100
105 110Ile Arg Ala Val Thr Trp Met Asp Asn
Pro Ile Glu Val Tyr Val Asn 115 120
125Asp Ser Val Trp Val Pro Gly Pro Ile Asp Asp Arg Cys Pro Ala Lys
130 135 140Pro Glu Glu Glu Gly Met Met
Ile Asn Ile Ser Ile Gly Tyr Arg Tyr145 150
155 160Pro Pro Ile Cys Leu Gly Arg Ala Pro Gly Cys Leu
Met Pro Ala Val 165 170
175Gln Asn Trp Leu Val Glu Val Pro Thr Val Ser Pro Ile Ser Arg Phe
180 185 190Thr Tyr His Met Val Ser
Gly Met Ser Leu Arg Pro Arg Val Asn Tyr 195 200
205Leu Gln Asp Phe Ser Tyr Gln Arg Ser Leu Lys Phe Arg Pro
Lys Gly 210 215 220Lys Pro Cys Pro Lys
Glu Ile Pro Lys Glu Ser Lys Asn Thr Glu Val225 230
235 240Leu Val Trp Glu Glu Cys Val Ala Asn Ser
Ala Val Ile Leu Gln Asn 245 250
255Asn Glu Phe Gly Thr Ile Ile Asp Trp Ala Pro Arg Gly Gln Phe Tyr
260 265 270His Asn Cys Ser Gly
Gln Thr Gln Ser Cys Pro Ser Ala Gln Val Ser 275
280 285Pro Ala Val Asp Ser Asp Leu Thr Glu Ser Leu Asp
Lys His Lys His 290 295 300Lys Lys Leu
Gln Ser Phe Tyr Pro Trp Glu Trp Gly Glu Lys Arg Ile305
310 315 320Ser Thr Pro Arg Pro Lys Ile
Val Ser Pro Val Ser Gly Pro Glu His 325
330 335Pro Glu Leu Trp Arg Leu Thr Val Ala Ser His His
Ile Arg Ile Trp 340 345 350Ser
Gly Asn Gln Thr Leu Glu Thr Arg Asp Arg Lys Pro Phe Tyr Thr 355
360 365Val Asp Leu Asn Ser Ser Leu Thr Leu
Pro Leu Gln Ser Cys Val Lys 370 375
380Pro Pro Tyr Met Leu Val Val Gly Asn Ile Val Ile Lys Pro Asp Ser385
390 395 400Gln Thr Ile Thr
Cys Glu Asn Cys Arg Leu Leu Thr Cys Ile Asp Ser 405
410 415Thr Phe Asn Trp Gln His Arg Ile Leu Leu
Val Arg Ala Arg Glu Gly 420 425
430Val Trp Ile Pro Val Ser Met Asp Arg Pro Trp Glu Ala Ser Pro Ser
435 440 445Val His Ile Leu Thr Glu Val
Leu Lys Gly Val Leu Asn Arg Ser Lys 450 455
460Arg Phe Ile Phe Thr Leu Ile Ala Val Ile Met Gly Leu Ile Ala
Val465 470 475 480Thr Ala
Thr Ala Ala Val Ala Gly Val Ala Leu His Ser Ser Val Gln
485 490 495Ser Val Asn Phe Val Asn Asp
Gly Gln Lys Asn Ser Thr Arg Leu Trp 500 505
510Asn Ser Gln Ser Ser Ile Asp Gln Lys Leu Ala Asn Gln Ile
Asn Asp 515 520 525Leu Arg Gln Thr
Val Ile Trp Met Gly Asp Arg Leu Met Ser Leu Glu 530
535 540His Arg Phe Gln Leu Gln Cys Asp Trp Asn Thr Ser
Asp Phe Cys Ile545 550 555
560Thr Pro Gln Ile Tyr Asn Asp Ser Glu His His Trp Asp Met Val Arg
565 570 575Arg His Leu Gln Gly
Arg Glu Asp Asn Leu Thr Leu Asp Ile Ser Lys 580
585 590Leu Lys Glu Gln Ile Phe Glu Ala Ser Lys Ala His
Leu Asn Leu Val 595 600 605Pro Gly
Thr Glu Ala Ile Ala Gly Val Ala Asp Gly Leu Ala Asn Leu 610
615 620Asn Pro Val Thr Trp Val Lys Thr Ile Gly Ser
Thr Thr Ile Ile Asn625 630 635
640Leu Ile Leu Ile Leu Val Cys Leu Phe Cys Leu Leu Leu Val Cys Arg
645 650 655Cys Thr Gln Gln
Leu Arg Arg Asp Ser Asp His Arg Glu Arg Ala Met 660
665 670Met Thr Met Ala Val Leu Ser Lys Arg Lys Gly
Gly Asn Val Gly Lys 675 680 685Ser
Lys Arg Asp Gln Ile Val Thr Val Ser Val 690
69511626PRTHomo sapiens 11Met Gly Pro Glu Ala Trp Val Arg Pro Leu Lys Thr
Ala Pro Lys Pro1 5 10
15Gly Glu Ala Ile Arg Leu Ile Leu Phe Ile Tyr Leu Ser Cys Phe Phe
20 25 30Leu Pro Val Met Ser Ser Glu
Pro Ser Tyr Ser Phe Leu Leu Thr Ser 35 40
45Phe Thr Thr Gly Arg Val Phe Ala Asn Thr Thr Trp Arg Ala Gly
Thr 50 55 60Ser Lys Glu Val Ser Phe
Ala Val Asp Leu Cys Val Leu Phe Pro Glu65 70
75 80Pro Ala Arg Thr His Glu Glu Gln His Asn Leu
Pro Val Ile Gly Ala 85 90
95Gly Ser Val Asp Leu Ala Ala Gly Phe Gly His Ser Gly Ser Gln Thr
100 105 110Gly Cys Gly Ser Ser Lys
Gly Ala Glu Lys Gly Leu Gln Asn Val Asp 115 120
125Phe Tyr Leu Cys Pro Gly Asn His Pro Asp Ala Ser Cys Arg
Asp Thr 130 135 140Tyr Gln Phe Phe Cys
Pro Asp Trp Thr Cys Val Thr Leu Ala Thr Tyr145 150
155 160Ser Gly Gly Ser Thr Arg Ser Ser Thr Leu
Ser Ile Ser Arg Val Pro 165 170
175His Pro Lys Leu Cys Thr Arg Lys Asn Cys Asn Pro Leu Thr Ile Thr
180 185 190Val His Asp Pro Asn
Ala Ala Gln Trp Tyr Tyr Gly Met Ser Trp Gly 195
200 205Leu Arg Leu Tyr Ile Pro Gly Phe Asp Val Gly Thr
Met Phe Thr Ile 210 215 220Gln Lys Lys
Ile Leu Val Ser Trp Ser Ser Pro Lys Pro Ile Gly Pro225
230 235 240Leu Thr Asp Leu Gly Asp Pro
Ile Phe Gln Lys His Pro Asp Lys Val 245
250 255Asp Leu Thr Val Pro Leu Pro Phe Leu Val Pro Arg
Pro Gln Leu Gln 260 265 270Gln
Gln His Leu Gln Pro Ser Leu Met Ser Ile Leu Gly Gly Val His 275
280 285His Leu Leu Asn Leu Thr Gln Pro Lys
Leu Ala Gln Asp Cys Trp Leu 290 295
300Cys Leu Lys Ala Lys Pro Pro Tyr Tyr Val Gly Leu Gly Val Glu Ala305
310 315 320Thr Leu Lys Arg
Gly Pro Leu Ser Cys His Thr Arg Pro Arg Ala Leu 325
330 335Thr Ile Gly Asp Val Ser Gly Asn Ala Ser
Cys Leu Ile Ser Thr Gly 340 345
350Tyr Asn Leu Ser Ala Ser Pro Phe Gln Ala Thr Cys Asn Gln Ser Leu
355 360 365Leu Thr Ser Ile Ser Thr Ser
Val Ser Tyr Gln Ala Pro Asn Asn Thr 370 375
380Trp Leu Ala Cys Thr Ser Gly Leu Thr Arg Cys Ile Asn Gly Thr
Glu385 390 395 400Pro Gly
Pro Leu Leu Cys Val Leu Val His Val Leu Pro Gln Val Tyr
405 410 415Val Tyr Ser Gly Pro Glu Gly
Arg Gln Leu Ile Ala Pro Pro Glu Leu 420 425
430His Pro Arg Leu His Gln Ala Val Pro Leu Leu Val Pro Leu
Leu Ala 435 440 445Gly Leu Ser Ile
Ala Gly Ser Ala Ala Ile Gly Thr Ala Ala Leu Val 450
455 460Gln Gly Glu Thr Gly Leu Ile Ser Leu Ser Gln Gln
Val Asp Ala Asp465 470 475
480Phe Ser Asn Leu Gln Ser Ala Ile Asp Ile Leu His Ser Gln Val Glu
485 490 495Ser Leu Ala Glu Val
Val Leu Gln Asn Cys Arg Cys Leu Asp Leu Leu 500
505 510Phe Leu Ser Gln Gly Gly Leu Cys Ala Ala Leu Gly
Glu Ser Cys Cys 515 520 525Phe Tyr
Ala Asn Gln Ser Gly Val Ile Lys Gly Thr Val Lys Lys Val 530
535 540Arg Glu Asn Leu Asp Arg His Gln Gln Glu Arg
Glu Asn Asn Ile Pro545 550 555
560Trp Tyr Gln Ser Met Phe Asn Trp Asn Pro Trp Leu Thr Thr Leu Ile
565 570 575Thr Gly Leu Ala
Gly Pro Leu Leu Ile Leu Leu Leu Ser Leu Ile Phe 580
585 590Gly Pro Cys Ile Leu Asn Ser Phe Leu Asn Phe
Ile Lys Gln Arg Ile 595 600 605Ala
Ser Val Lys Leu Thr Tyr Leu Lys Thr Gln Tyr Asp Thr Leu Val 610
615 620Asn Asn62512538PRTHomo sapiens 12Met Ala
Leu Pro Tyr His Ile Phe Leu Phe Thr Val Leu Leu Pro Ser1 5
10 15Phe Thr Leu Thr Ala Pro Pro Pro
Cys Arg Cys Met Thr Ser Ser Ser 20 25
30Pro Tyr Gln Glu Phe Leu Trp Arg Met Gln Arg Pro Gly Asn Ile
Asp 35 40 45Ala Pro Ser Tyr Arg
Ser Leu Ser Lys Gly Thr Pro Thr Phe Thr Ala 50 55
60His Thr His Met Pro Arg Asn Cys Tyr His Ser Ala Thr Leu
Cys Met65 70 75 80His
Ala Asn Thr His Tyr Trp Thr Gly Lys Met Ile Asn Pro Ser Cys
85 90 95Pro Gly Gly Leu Gly Val Thr
Val Cys Trp Thr Tyr Phe Thr Gln Thr 100 105
110Gly Met Ser Asp Gly Gly Gly Val Gln Asp Gln Ala Arg Glu
Lys His 115 120 125Val Lys Glu Val
Ile Ser Gln Leu Thr Arg Val His Gly Thr Ser Ser 130
135 140Pro Tyr Lys Gly Leu Asp Leu Ser Lys Leu His Glu
Thr Leu Arg Thr145 150 155
160His Thr Arg Leu Val Ser Leu Phe Asn Thr Thr Leu Thr Gly Leu His
165 170 175Glu Val Ser Ala Gln
Asn Pro Thr Asn Cys Trp Ile Cys Leu Pro Leu 180
185 190Asn Phe Arg Pro Tyr Val Ser Ile Pro Val Pro Glu
Gln Trp Asn Asn 195 200 205Phe Ser
Thr Glu Ile Asn Thr Thr Ser Val Leu Val Gly Pro Leu Val 210
215 220Ser Asn Leu Glu Ile Thr His Thr Ser Asn Leu
Thr Cys Val Lys Phe225 230 235
240Ser Asn Thr Thr Tyr Thr Thr Asn Ser Gln Cys Ile Arg Trp Val Thr
245 250 255Pro Pro Thr Gln
Ile Val Cys Leu Pro Ser Gly Ile Phe Phe Val Cys 260
265 270Gly Thr Ser Ala Tyr Arg Cys Leu Asn Gly Ser
Ser Glu Ser Met Cys 275 280 285Phe
Leu Ser Phe Leu Val Pro Pro Met Thr Ile Tyr Thr Glu Gln Asp 290
295 300Leu Tyr Ser Tyr Val Ile Ser Lys Pro Arg
Asn Lys Arg Val Pro Ile305 310 315
320Leu Pro Phe Val Ile Gly Ala Gly Val Leu Gly Ala Leu Gly Thr
Gly 325 330 335Ile Gly Gly
Ile Thr Thr Ser Thr Gln Phe Tyr Tyr Lys Leu Ser Gln 340
345 350Glu Leu Asn Gly Asp Met Glu Arg Val Ala
Asp Ser Leu Val Thr Leu 355 360
365Gln Asp Gln Leu Asn Ser Leu Ala Ala Val Val Leu Gln Asn Arg Arg 370
375 380Ala Leu Asp Leu Leu Thr Ala Glu
Arg Gly Gly Thr Cys Leu Phe Leu385 390
395 400Gly Glu Glu Cys Cys Tyr Tyr Val Asn Gln Ser Gly
Ile Val Thr Glu 405 410
415Lys Val Lys Glu Ile Arg Asp Arg Ile Gln Arg Arg Ala Glu Glu Leu
420 425 430Arg Asn Thr Gly Pro Trp
Gly Leu Leu Ser Gln Trp Met Pro Trp Ile 435 440
445Leu Pro Phe Leu Gly Pro Leu Ala Ala Ile Ile Leu Leu Leu
Leu Phe 450 455 460Gly Pro Cys Ile Phe
Asn Leu Leu Val Asn Phe Val Ser Ser Arg Ile465 470
475 480Glu Ala Val Lys Leu Gln Met Glu Pro Lys
Met Gln Ser Lys Thr Lys 485 490
495Ile Tyr Arg Arg Pro Leu Asp Arg Pro Ala Ser Pro Arg Ser Asp Val
500 505 510Asn Asp Ile Lys Gly
Thr Pro Pro Glu Glu Ile Ser Ala Ala Gln Pro 515
520 525Leu Leu Arg Pro Asn Ser Ala Gly Ser Ser 530
53513538PRTHomo sapiens 13Met Gly Leu Leu Leu Leu Val Leu Ile
Leu Thr Pro Ser Leu Ala Ala1 5 10
15Tyr Arg His Pro Asp Phe Pro Leu Leu Glu Lys Ala Gln Gln Leu
Leu 20 25 30Gln Ser Thr Gly
Ser Pro Tyr Ser Thr Asn Cys Trp Leu Cys Thr Ser 35
40 45Ser Ser Thr Glu Thr Pro Gly Thr Ala Tyr Pro Ala
Ser Pro Arg Glu 50 55 60Trp Thr Ser
Ile Glu Ala Glu Leu His Ile Ser Tyr Arg Trp Asp Pro65 70
75 80Asn Leu Lys Gly Leu Met Arg Pro
Ala Asn Ser Leu Leu Ser Thr Val 85 90
95Lys Gln Asp Phe Pro Asp Ile Arg Gln Lys Pro Pro Ile Phe
Gly Pro 100 105 110Ile Phe Thr
Asn Ile Asn Leu Met Gly Ile Ala Pro Ile Cys Val Met 115
120 125Ala Lys Arg Lys Asn Gly Thr Asn Val Gly Thr
Leu Pro Ser Thr Val 130 135 140Cys Asn
Val Thr Phe Thr Val Asp Ser Asn Gln Gln Thr Tyr Gln Thr145
150 155 160Tyr Thr His Asn Gln Phe Arg
His Gln Pro Arg Phe Pro Lys Pro Pro 165
170 175Asn Ile Thr Phe Pro Gln Gly Thr Leu Leu Asp Lys
Ser Ser Arg Phe 180 185 190Cys
Gln Gly Arg Pro Ser Ser Cys Ser Thr Arg Asn Phe Trp Phe Arg 195
200 205Pro Ala Asp Tyr Asn Gln Cys Leu Gln
Ile Ser Asn Leu Ser Ser Thr 210 215
220Ala Glu Trp Val Leu Leu Asp Gln Thr Arg Asn Ser Leu Phe Trp Glu225
230 235 240Asn Lys Thr Lys
Gly Ala Asn Gln Ser Gln Thr Pro Cys Val Gln Val 245
250 255Leu Ala Gly Met Thr Ile Ala Thr Ser Tyr
Leu Gly Ile Ser Ala Val 260 265
270Ser Glu Phe Phe Gly Thr Ser Leu Thr Pro Leu Phe His Phe His Ile
275 280 285Ser Thr Cys Leu Lys Thr Gln
Gly Ala Phe Tyr Ile Cys Gly Gln Ser 290 295
300Ile His Gln Cys Leu Pro Ser Asn Trp Thr Gly Thr Cys Thr Ile
Gly305 310 315 320Tyr Val
Thr Pro Asp Ile Phe Ile Ala Pro Gly Asn Leu Ser Leu Pro
325 330 335Ile Pro Ile Tyr Gly Asn Ser
Pro Leu Pro Arg Val Arg Arg Ala Ile 340 345
350His Phe Ile Pro Leu Leu Ala Gly Leu Gly Ile Leu Ala Gly
Thr Gly 355 360 365Thr Gly Ile Ala
Gly Ile Thr Lys Ala Ser Leu Thr Tyr Ser Gln Leu 370
375 380Ser Lys Glu Ile Ala Asn Asn Ile Asp Thr Met Ala
Lys Ala Leu Thr385 390 395
400Thr Met Gln Glu Gln Ile Asp Ser Leu Ala Ala Val Val Leu Gln Asn
405 410 415Arg Arg Gly Leu Asp
Met Leu Thr Ala Ala Gln Gly Gly Ile Cys Leu 420
425 430Ala Leu Asp Glu Lys Cys Cys Phe Trp Val Asn Gln
Ser Gly Lys Val 435 440 445Gln Asp
Asn Ile Arg Gln Leu Leu Asn Gln Ala Ser Ser Leu Arg Glu 450
455 460Arg Ala Thr Gln Gly Trp Leu Asn Trp Glu Gly
Thr Trp Lys Trp Phe465 470 475
480Ser Trp Val Leu Pro Leu Thr Gly Pro Leu Val Ser Leu Leu Leu Leu
485 490 495Leu Leu Phe Gly
Pro Cys Leu Leu Asn Leu Ile Thr Gln Phe Val Ser 500
505 510Ser Arg Leu Gln Ala Ile Lys Leu Gln Thr Asn
Leu Ser Ala Gly Arg 515 520 525His
Pro Arg Asn Ile Gln Glu Ser Pro Phe 530
53514604PRTHomo sapiens 14Met Leu Gly Met Asn Met Leu Leu Ile Thr Leu Phe
Leu Leu Leu Pro1 5 10
15Leu Ser Met Leu Lys Gly Glu Pro Trp Glu Gly Cys Leu His Cys Thr
20 25 30His Thr Thr Trp Ser Gly Asn
Ile Met Thr Lys Thr Leu Leu Tyr His 35 40
45Thr Tyr Tyr Glu Cys Ala Gly Thr Cys Leu Gly Thr Cys Thr His
Asn 50 55 60Gln Thr Thr Tyr Ser Val
Cys Asp Pro Gly Arg Gly Gln Pro Tyr Val65 70
75 80Cys Tyr Asp Pro Lys Ser Ser Pro Gly Thr Trp
Phe Glu Ile His Val 85 90
95Gly Ser Lys Glu Gly Asp Leu Leu Asn Gln Thr Lys Val Phe Pro Ser
100 105 110Gly Lys Asp Val Val Ser
Leu Tyr Phe Asp Val Cys Gln Ile Val Ser 115 120
125Met Gly Ser Leu Phe Pro Val Ile Phe Ser Ser Met Glu Tyr
Tyr Ser 130 135 140Ser Cys His Lys Asn
Arg Tyr Ala His Pro Ala Cys Ser Thr Asp Ser145 150
155 160Pro Val Thr Thr Cys Trp Asp Cys Thr Thr
Trp Ser Thr Asn Gln Gln 165 170
175Ser Leu Gly Pro Ile Met Leu Thr Lys Ile Pro Leu Glu Pro Asp Cys
180 185 190Lys Thr Ser Thr Cys
Asn Ser Val Asn Leu Thr Ile Leu Glu Pro Asp 195
200 205Gln Pro Ile Trp Thr Thr Gly Leu Lys Ala Pro Leu
Gly Ala Arg Val 210 215 220Ser Gly Glu
Glu Ile Gly Pro Gly Ala Tyr Val Tyr Leu Tyr Ile Ile225
230 235 240Lys Lys Thr Arg Thr Arg Ser
Thr Gln Gln Phe Arg Val Phe Glu Ser 245
250 255Phe Tyr Glu His Val Asn Gln Lys Leu Pro Glu Pro
Pro Pro Leu Ala 260 265 270Ser
Asn Leu Phe Ala Gln Leu Ala Glu Asn Ile Ala Ser Ser Leu His 275
280 285Val Ala Ser Cys Tyr Val Cys Gly Gly
Met Asn Met Gly Asp Gln Trp 290 295
300Pro Trp Glu Ala Arg Glu Leu Met Pro Gln Asp Asn Phe Thr Leu Thr305
310 315 320Ala Ser Ser Leu
Glu Pro Ala Pro Ser Ser Gln Ser Ile Trp Phe Leu 325
330 335Lys Thr Ser Ile Ile Gly Lys Phe Cys Ile
Ala Arg Trp Gly Lys Ala 340 345
350Phe Thr Asp Pro Val Gly Glu Leu Thr Cys Leu Gly Gln Gln Tyr Tyr
355 360 365Asn Glu Thr Leu Gly Lys Thr
Leu Trp Arg Gly Lys Ser Asn Asn Ser 370 375
380Glu Ser Pro His Pro Ser Pro Phe Ser Arg Phe Pro Ser Leu Asn
His385 390 395 400Ser Trp
Tyr Gln Leu Glu Ala Pro Asn Thr Trp Gln Ala Pro Ser Gly
405 410 415Leu Tyr Trp Ile Cys Gly Pro
Gln Ala Tyr Arg Gln Leu Pro Ala Lys 420 425
430Trp Ser Gly Ala Cys Val Leu Gly Thr Ile Arg Pro Ser Phe
Phe Leu 435 440 445Met Pro Leu Lys
Gln Gly Glu Ala Leu Gly Tyr Pro Ile Tyr Asp Glu 450
455 460Thr Lys Arg Lys Ser Lys Arg Gly Ile Thr Ile Gly
Asp Trp Lys Asp465 470 475
480Asn Glu Trp Pro Pro Glu Arg Ile Ile Gln Tyr Tyr Gly Pro Ala Thr
485 490 495Trp Ala Glu Asp Gly
Met Trp Gly Tyr Arg Thr Pro Val Tyr Met Leu 500
505 510Asn Arg Ile Ile Arg Leu Gln Ala Val Leu Glu Ile
Ile Thr Asn Glu 515 520 525Thr Ala
Gly Ala Leu Asn Leu Leu Ala Gln Gln Ala Thr Lys Met Arg 530
535 540Asn Val Ile Tyr Gln Asn Arg Leu Ala Leu Asp
Tyr Leu Leu Ala Gln545 550 555
560Glu Glu Gly Val Cys Gly Lys Phe Asn Leu Thr Asn Cys Cys Leu Glu
565 570 575Leu Asp Asp Glu
Gly Lys Val Ile Lys Glu Ile Thr Ala Lys Ile Gln 580
585 590Lys Leu Ala His Ile Pro Val Gln Thr Trp Lys
Gly 595 60015514PRTHomo sapiens 15Met Asp Pro Leu
His Thr Ile Glu Lys Val Pro Ala Arg Arg Asn Ile1 5
10 15His Asp Arg Gly His Gln Gly His Arg Met
Gly Asp Gly Thr Pro Gly 20 25
30Arg Pro Lys Ile Ser Val Gln Gln Met Thr Arg Phe Ser Leu Ile Ile
35 40 45Phe Phe Leu Ser Ala Pro Phe Val
Val Asn Ala Ser Thr Ser Asn Val 50 55
60Phe Leu Gln Trp Ala His Ser Tyr Ala Asp Gly Leu Gln Gln Gly Asp65
70 75 80Pro Cys Trp Val Cys
Gly Ser Leu Pro Val Thr Asn Thr Met Glu Leu 85
90 95Pro Trp Trp Val Ser Pro Leu Gln Gly Lys Asp
Trp Val Phe Phe Gln 100 105
110Ser Phe Ile Gly Asp Leu Lys Gln Trp Thr Gly Ala Gln Met Thr Gly
115 120 125Val Thr Arg Lys Asn Ile Ser
Glu Trp Pro Ile Asn Lys Thr Leu Asn 130 135
140Glu Pro Gly His Asp Lys Pro Phe Ser Val Asn Glu Thr Arg Asp
Lys145 150 155 160Val Ile
Ala Phe Ala Ile Pro Leu Leu Asp Thr Lys Val Phe Val Gln
165 170 175Thr Ser Arg Pro Gln Asn Thr
Gln Tyr Arg Asn Gly Phe Leu Gln Ile 180 185
190Trp Asp Gly Phe Ile Trp Leu Thr Ala Thr Lys Gly His Leu
Ser Gln 195 200 205Ile Ala Pro Leu
Cys Trp Glu Gln Arg Asn His Ser Leu Asp Asn Trp 210
215 220Pro Asn Thr Thr Arg Val Met Gly Trp Ile Pro Pro
Gly Gln Cys Arg225 230 235
240His Thr Ile Leu Leu Gln Gln Arg Asp Leu Phe Ala Thr Asp Trp Ser
245 250 255Gln Gln Pro Gly Leu
Asn Trp Tyr Ala Pro Asn Gly Thr Gln Trp Leu 260
265 270Cys Ser Pro Asn Leu Trp Pro Trp Leu Pro Ser Gly
Trp Leu Gly Cys 275 280 285Cys Thr
Leu Gly Ile Pro Trp Ala Gln Gly Arg Trp Val Lys Thr Met 290
295 300Glu Val Tyr Pro Tyr Leu Pro His Val Val Asn
Gln Gly Thr Arg Ala305 310 315
320Ile Val His Arg Asn Asp His Leu Pro Thr Ile Phe Met Pro Ser Val
325 330 335Gly Leu Gly Thr
Val Ile Gln His Ile Glu Ala Leu Ala Asn Phe Thr 340
345 350Gln Arg Ala Leu Asn Asp Ser Leu Gln Ser Ile
Ser Leu Met Asn Ala 355 360 365Glu
Val Tyr Tyr Met His Glu Asp Ile Leu Gln Asn Arg Met Ala Leu 370
375 380Asp Ile Leu Thr Ala Ala Glu Gly Gly Thr
Cys Ala Leu Ile Lys Thr385 390 395
400Glu Cys Cys Val Tyr Ile Pro Asn Asn Ser Arg Asn Ile Ser Leu
Ala 405 410 415Leu Glu Asp
Thr Cys Arg Gln Ile Gln Val Ile Ser Ser Ser Ala Leu 420
425 430Ser Leu His Asp Trp Ile Ala Ser Gln Phe
Ser Gly Arg Pro Ser Trp 435 440
445Trp Gln Lys Ile Leu Ile Val Leu Ala Thr Leu Trp Ser Val Gly Ile 450
455 460Ala Leu Cys Cys Gly Leu Tyr Phe
Cys Arg Met Phe Ser Gln His Ile465 470
475 480Pro Gln Thr His Ser Ile Ile Phe Gln Gln Glu Leu
Pro Leu Ser Pro 485 490
495Pro Ser Gln Glu His Tyr Gln Ser Gln Arg Asp Ile Phe His Ser Asn
500 505 510Ala Pro16527PRTHomo
sapiens 16Met Asn Ser Pro Cys Asp Arg Leu Gln Gln Phe Ile Gln Val Leu
Leu1 5 10 15Glu Glu Ser
Trp Ser Phe Pro Ser Phe Ala Asn Thr Leu His Trp Pro 20
25 30Glu Asn Leu Leu Ser Tyr Ile Asp Glu Leu
Val Trp Gln Gly Ser Leu 35 40
45Gln Asn Phe His Gln His Glu Val Arg Phe Asp Lys Pro Pro Leu Arg 50
55 60Leu Pro Leu Thr Gly Phe Ser Ser Leu
Thr Glu Asn Trp Ser Ser Arg65 70 75
80Gln Ala Val Ser Ser Arg Leu Val Ala Thr Ala Ala Ser Pro
Pro Ala 85 90 95Gly Cys
Gln Ala Pro Ile Ala Phe Leu Gly Leu Lys Phe Ser Ser Leu 100
105 110Gly Pro Ala Arg Lys Asn Pro Ala Leu
Cys Phe Leu Tyr Asp Gln Ser 115 120
125Asn Ser Lys Cys Asn Thr Ser Trp Val Lys Glu Asn Val Gly Cys Pro
130 135 140Trp His Trp Cys Asn Ile His
Glu Ala Leu Ile Arg Thr Glu Lys Gly145 150
155 160Ser Asp Pro Met Phe Tyr Val Asn Thr Ser Thr Gly
Gly Arg Asp Gly 165 170
175Phe Asn Gly Phe Asn Leu Gln Ile Ser Asp Pro Trp Asp Pro Arg Trp
180 185 190Ala Ser Gly Val Asp Gly
Gly Leu Tyr Glu His Lys Thr Phe Met Tyr 195 200
205Pro Val Ala Lys Ile Arg Ile Ala Arg Thr Leu Lys Thr Thr
Val Thr 210 215 220Gly Leu Ser Asp Leu
Ala Ser Ser Ile Gln Ser Ala Glu Lys Glu Leu225 230
235 240Thr Ser Gln Leu Gln Pro Ala Ala Asp Gln
Ala Lys Ser Ser Arg Phe 245 250
255Ser Trp Leu Thr Leu Ile Ser Glu Gly Ala Gln Leu Leu Gln Ser Thr
260 265 270Gly Val Gln Asn Leu
Ser His Cys Phe Leu Cys Ala Ala Leu Arg Arg 275
280 285Pro Pro Leu Val Ala Val Pro Leu Pro Thr Pro Phe
Asn Tyr Thr Ile 290 295 300Asn Ser Ser
Thr Pro Ile Pro Pro Val Pro Lys Gly Gln Val Pro Leu305
310 315 320Phe Ser Asp Pro Ile Arg His
Lys Phe Pro Phe Cys Tyr Ser Thr Pro 325
330 335Asn Ala Ser Trp Cys Asn Gln Thr Arg Met Leu Thr
Ser Thr Pro Ala 340 345 350Pro
Pro Arg Gly Tyr Phe Trp Cys Asn Ser Thr Leu Thr Lys Val Leu 355
360 365Asn Ser Thr Gly Asn His Thr Leu Cys
Leu Pro Ile Ser Leu Ile Pro 370 375
380Gly Leu Thr Leu Tyr Ser Gln Asp Glu Leu Ser His Leu Leu Ala Trp385
390 395 400Thr Glu Pro Arg
Pro Gln Asn Lys Ser Lys Trp Ala Ile Phe Leu Pro 405
410 415Leu Val Leu Gly Ile Ser Leu Ala Ser Ser
Leu Val Ala Ser Gly Leu 420 425
430Gly Lys Gly Ala Leu Thr His Ser Ile Gln Thr Ser Gln Asp Leu Ser
435 440 445Thr His Leu Gln Leu Ala Ile
Glu Ala Ser Ala Glu Ser Leu Asp Ser 450 455
460Leu Gln Arg Gln Ile Thr Thr Val Ala Gln Val Ala Ala Gln Asn
Arg465 470 475 480Gln Ala
Leu Asp Leu Leu Met Ala Glu Lys Gly Arg Thr Cys Leu Phe
485 490 495Leu Gln Glu Glu Cys Cys Tyr
Tyr Leu Asn Glu Ser Gly Val Val Glu 500 505
510Asn Ser Leu Gln Thr Leu Lys Lys Lys Lys Ser Ser Lys Arg
Ser 515 520 52517584PRTHomo
sapiens 17Met Ala Arg Pro Ser Pro Leu Cys Leu Leu Leu Leu Leu Thr Leu
Leu1 5 10 15Thr Pro Ile
Val Pro Ser Asn Ser Leu Leu Thr Glu Pro Pro Phe Arg 20
25 30Trp Arg Phe Tyr Leu His Glu Thr Trp Thr
Gln Gly Asn Arg Leu Ser 35 40
45Thr Val Thr Leu Ala Thr Val Asp Cys Gln Pro His Gly Cys Gln Ala 50
55 60Gln Val Thr Phe Asn Phe Thr Ser Phe
Lys Ser Val Leu Arg Gly Trp65 70 75
80Ser Asn Pro Thr Ile Cys Phe Val Tyr Asp Gln Thr His Ser
Asn Cys 85 90 95Arg Asp
Tyr Trp Val Asp Thr Asn Gly Gly Cys Pro Tyr Ala Tyr Cys 100
105 110Arg Met His Val Thr Gln Leu His Thr
Ala Lys Lys Leu Gln His Thr 115 120
125Tyr Arg Leu Thr Ser Asp Gly Arg Thr Thr Tyr Phe Leu Thr Ile Pro
130 135 140Asp Pro Trp Asp Ser Arg Trp
Val Ser Gly Val Thr Gly Arg Leu Tyr145 150
155 160Arg Trp Pro Thr Asp Ser Tyr Pro Val Gly Lys Leu
Arg Ile Phe Leu 165 170
175Thr Tyr Ile Arg Val Ile Pro Gln Val Leu Ser Asn Leu Lys Asp Gln
180 185 190Ala Asp Asn Ile Lys His
Gln Glu Glu Val Ile Asn Thr Leu Val Gln 195 200
205Ser His Pro Lys Ala Asp Met Val Thr Tyr Asp Asp Lys Ala
Glu Ala 210 215 220Gly Pro Phe Ser Trp
Ile Thr Leu Val Arg His Gly Ala Arg Leu Val225 230
235 240Asn Met Ala Gly Leu Val Asn Leu Ser His
Cys Phe Leu Cys Thr Ala 245 250
255Leu Ser Gln Pro Pro Leu Val Ala Val Pro Leu Pro Gln Ala Phe Asn
260 265 270Thr Ser Gly Asn His
Thr Ala His Pro Ser Gly Val Phe Ser Glu Gln 275
280 285Val Pro Leu Phe Arg Asp Pro Leu Gln Pro Gln Phe
Pro Phe Cys Tyr 290 295 300Thr Thr Pro
Asn Ser Ser Trp Cys Asn Gln Thr Tyr Ser Gly Ser Leu305
310 315 320Ser Asn Leu Ser Ala Pro Ala
Gly Gly Tyr Phe Trp Cys Asn Phe Thr 325
330 335Leu Thr Lys His Leu Asn Ile Ser Ser Asn Asn Thr
Leu Ser Arg Asn 340 345 350Leu
Cys Leu Pro Ile Ser Leu Val Pro Arg Leu Thr Leu Tyr Ser Glu 355
360 365Ala Glu Leu Ser Ser Leu Val Asn Pro
Pro Met Arg Gln Lys Arg Ala 370 375
380Val Phe Pro Pro Leu Val Ile Gly Val Ser Leu Thr Ser Ser Leu Val385
390 395 400Ala Ser Gly Leu
Gly Thr Gly Ala Ile Val His Phe Ile Ser Ser Ser 405
410 415Gln Asp Leu Ser Ile Lys Leu Gln Met Ala
Ile Glu Ala Ser Ala Glu 420 425
430Ser Leu Ala Ser Leu Gln Arg Gln Ile Thr Ser Val Ala Lys Val Ala
435 440 445Met Gln Asn Arg Arg Ala Leu
Asp Leu Leu Thr Ala Asp Lys Gly Gly 450 455
460Thr Cys Met Phe Leu Gly Glu Glu Cys Cys Tyr Tyr Ile Asn Glu
Ser465 470 475 480Gly Leu
Val Glu Thr Ser Leu Leu Thr Leu Asp Lys Ile Arg Asp Gly
485 490 495Leu His Arg Pro Ser Ser Thr
Pro Asn Tyr Gly Gly Gly Trp Trp Gln 500 505
510Ser Pro Leu Thr Thr Trp Ile Ile Pro Phe Ile Ser Pro Ile
Leu Ile 515 520 525Ile Cys Leu Leu
Leu Leu Ile Ala Pro Cys Val Leu Lys Phe Ile Lys 530
535 540Asn Arg Ile Ser Glu Val Ser Arg Val Thr Val Asn
Gln Met Leu Leu545 550 555
560His Pro Tyr Ser Arg Leu Pro Thr Ser Glu Asp His Tyr Asp Asp Ala
565 570 575Leu Thr Gln Gln Glu
Ala Ala Arg 58018699PRTArtificial SequenceDescription of
Artificial Sequence Synthetic polypeptide 18Met Asn Pro Ser Glu Met
Gln Arg Lys Ala Pro Pro Arg Arg Arg Arg1 5
10 15His Arg Asn Arg Ala Pro Leu Thr His Lys Met Asn
Lys Met Val Thr 20 25 30Ser
Glu Glu Gln Met Lys Leu Pro Ser Thr Lys Lys Ala Glu Pro Pro 35
40 45Thr Trp Ala Gln Leu Lys Lys Leu Thr
Gln Leu Ala Thr Lys Tyr Leu 50 55
60Glu Asn Thr Lys Val Thr Gln Thr Pro Glu Ser Met Leu Leu Ala Ala65
70 75 80Leu Met Ile Val Ser
Met Val Val Ser Leu Pro Met Pro Ala Gly Ala 85
90 95Ala Ala Ala Asn Tyr Thr Tyr Trp Ala Tyr Val
Pro Phe Pro Pro Leu 100 105
110Ile Arg Ala Val Thr Trp Met Asp Asn Pro Ile Glu Val Tyr Val Asn
115 120 125Asp Ser Val Trp Val Pro Gly
Pro Ile Asp Asp Arg Cys Pro Ala Lys 130 135
140Pro Glu Glu Glu Gly Met Met Ile Asn Ile Ser Ile Gly Tyr Arg
Tyr145 150 155 160Pro Pro
Ile Cys Leu Gly Arg Ala Pro Gly Cys Leu Met Pro Ala Val
165 170 175Gln Asn Trp Leu Val Glu Val
Pro Thr Val Ser Pro Ile Ser Arg Phe 180 185
190Thr Tyr His Met Val Ser Gly Met Ser Leu Arg Pro Arg Val
Asn Tyr 195 200 205Leu Gln Asp Phe
Ser Tyr Gln Arg Ser Leu Lys Phe Arg Pro Lys Gly 210
215 220Lys Pro Cys Pro Lys Glu Ile Pro Lys Glu Ser Lys
Asn Thr Glu Val225 230 235
240Leu Val Trp Glu Glu Cys Val Ala Asn Ser Ala Val Ile Leu Gln Asn
245 250 255Asn Glu Phe Gly Thr
Ile Ile Asp Trp Ala Pro Arg Gly Gln Phe Tyr 260
265 270His Asn Cys Ser Gly Gln Thr Gln Ser Cys Pro Ser
Ala Gln Val Ser 275 280 285Pro Ala
Val Asp Ser Asp Leu Thr Glu Ser Leu Asp Lys His Lys His 290
295 300Lys Lys Leu Gln Ser Phe Tyr Pro Trp Glu Trp
Gly Glu Lys Gly Ile305 310 315
320Ser Thr Pro Arg Pro Lys Ile Val Ser Pro Val Ser Gly Pro Glu His
325 330 335Pro Glu Leu Trp
Arg Leu Thr Val Ala Ser His His Ile Arg Ile Trp 340
345 350Ser Gly Asn Gln Thr Leu Glu Thr Arg Asp Arg
Lys Pro Phe Tyr Thr 355 360 365Val
Asp Leu Asn Ser Ser Leu Thr Val Pro Leu Gln Ser Cys Val Lys 370
375 380Pro Pro Tyr Met Leu Val Val Gly Asn Ile
Val Ile Lys Pro Asp Ser385 390 395
400Gln Thr Ile Thr Cys Glu Asn Cys Arg Leu Leu Thr Cys Ile Asp
Ser 405 410 415Thr Phe Asn
Trp Gln His Arg Ile Leu Leu Val Arg Ala Arg Glu Gly 420
425 430Val Trp Ile Pro Val Ser Met Asp Arg Pro
Trp Glu Ala Ser Pro Ser 435 440
445Val His Ile Leu Thr Glu Val Leu Lys Gly Val Leu Asn Arg Ser Lys 450
455 460Arg Phe Ile Phe Thr Leu Ile Ala
Val Ile Met Gly Leu Ile Ala Val465 470
475 480Thr Ala Thr Ala Ala Val Ala Gly Val Ala Leu His
Ser Ser Val Gln 485 490
495Ser Val Asn Phe Val Asn Asp Trp Gln Lys Asn Ser Thr Arg Leu Trp
500 505 510Asn Ser Gln Ser Ser Ile
Asp Gln Lys Leu Ala Asn Gln Ile Asn Asp 515 520
525Leu Arg Gln Thr Val Ile Trp Met Gly Asp Arg Leu Met Ser
Leu Glu 530 535 540His Arg Phe Gln Leu
Gln Cys Asp Trp Asn Thr Ser Asp Phe Cys Ile545 550
555 560Thr Pro Gln Ile Tyr Asn Glu Ser Glu His
His Trp Asp Met Val Arg 565 570
575Arg His Leu Gln Gly Arg Glu Asp Asn Leu Thr Leu Asp Ile Ser Lys
580 585 590Leu Lys Glu Gln Ile
Phe Glu Ala Ser Lys Ala His Leu Asn Leu Val 595
600 605Pro Gly Thr Glu Ala Ile Ala Gly Val Ala Asp Gly
Leu Ala Asn Leu 610 615 620Asn Pro Val
Thr Trp Val Lys Thr Ile Gly Ser Thr Thr Ile Ile Asn625
630 635 640Leu Ile Leu Ile Leu Val Cys
Leu Phe Cys Leu Leu Leu Val Cys Arg 645
650 655Cys Thr Gln Gln Leu Arg Arg Asp Ser Asp His Arg
Glu Arg Ala Met 660 665 670Met
Thr Met Ala Val Leu Ser Lys Arg Lys Gly Gly Asn Val Gly Lys 675
680 685Ser Lys Arg Asp Gln Ile Val Thr Val
Ser Val 690 69519666PRTArtificial SequenceDescription
of Artificial Sequence Synthetic polypeptide 19Met Gly Gln Thr Lys
Ser Lys Ile Lys Ser Lys Tyr Ala Ser Tyr Leu1 5
10 15Ser Phe Ile Lys Ile Leu Leu Lys Arg Gly Gly
Val Lys Val Ser Thr 20 25
30Lys Asn Leu Ile Lys Leu Phe Gln Ile Ile Glu Gln Phe Cys Pro Trp
35 40 45Phe Pro Glu Gln Gly Thr Leu Asp
Leu Lys Asp Trp Lys Arg Ile Gly 50 55
60Lys Glu Leu Lys Gln Ala Gly Arg Lys Gly Asn Ile Ile Pro Leu Thr65
70 75 80Val Trp Asn Asp Trp
Ala Ile Ile Lys Ala Ala Leu Glu Pro Phe Gln 85
90 95Thr Glu Glu Asp Ser Val Ser Val Ser Asp Ala
Pro Gly Ser Cys Ile 100 105
110Ile Asp Cys Asn Glu Asn Thr Arg Lys Lys Ser Gln Lys Glu Thr Glu
115 120 125Gly Leu His Cys Glu Tyr Val
Ala Glu Pro Val Met Ala Gln Ser Thr 130 135
140Gln Asn Val Asp Tyr Asn Gln Leu Gln Glu Val Ile Tyr Pro Glu
Thr145 150 155 160Leu Lys
Leu Glu Gly Lys Gly Pro Glu Leu Val Gly Pro Ser Glu Ser
165 170 175Lys Pro Arg Gly Thr Ser Pro
Leu Pro Ala Gly Gln Val Pro Val Thr 180 185
190Leu Gln Pro Gln Lys Gln Val Lys Glu Asn Lys Thr Gln Pro
Pro Val 195 200 205Ala Tyr Gln Tyr
Trp Pro Pro Ala Glu Leu Gln Tyr Arg Pro Pro Pro 210
215 220Glu Ser Gln Tyr Gly Tyr Pro Gly Met Pro Pro Ala
Pro Gln Gly Arg225 230 235
240Ala Pro Tyr Pro Gln Pro Pro Thr Arg Arg Leu Asn Pro Thr Ala Pro
245 250 255Pro Ser Arg Gln Gly
Ser Glu Leu His Glu Ile Ile Asp Lys Ser Arg 260
265 270Lys Glu Gly Asp Thr Glu Ala Trp Gln Phe Pro Val
Thr Leu Glu Pro 275 280 285Met Pro
Pro Gly Glu Gly Ala Gln Glu Gly Glu Pro Pro Thr Val Glu 290
295 300Ala Arg Tyr Lys Ser Phe Ser Ile Lys Met Leu
Lys Asp Met Lys Glu305 310 315
320Gly Val Lys Gln Tyr Gly Pro Asn Ser Pro Tyr Met Arg Thr Leu Leu
325 330 335Asp Ser Ile Ala
His Gly His Arg Leu Ile Pro Tyr Asp Trp Glu Ile 340
345 350Leu Ala Lys Ser Ser Leu Ser Pro Ser Gln Phe
Leu Gln Phe Lys Thr 355 360 365Trp
Trp Ile Asp Gly Val Gln Glu Gln Val Arg Arg Asn Arg Ala Ala 370
375 380Asn Pro Pro Val Asn Ile Asp Ala Asp Gln
Leu Leu Gly Ile Gly Gln385 390 395
400Asn Trp Ser Thr Ile Ser Gln Gln Ala Leu Met Gln Asn Glu Ala
Ile 405 410 415Glu Gln Val
Arg Ala Ile Cys Leu Arg Ala Trp Glu Lys Ile Gln Asp 420
425 430Pro Gly Ser Thr Cys Pro Ser Phe Asn Thr
Val Arg Gln Gly Ser Lys 435 440
445Glu Pro Tyr Pro Asp Phe Val Ala Arg Leu Gln Asp Val Ala Gln Lys 450
455 460Ser Ile Ala Asp Glu Lys Ala Arg
Lys Val Ile Val Glu Leu Met Ala465 470
475 480Tyr Glu Asn Ala Asn Pro Glu Cys Gln Ser Ala Ile
Lys Pro Leu Lys 485 490
495Gly Lys Val Pro Ala Gly Ser Asp Val Ile Ser Glu Tyr Val Lys Ala
500 505 510Cys Asp Gly Ile Gly Gly
Ala Met His Lys Ala Met Leu Met Ala Gln 515 520
525Ala Ile Thr Gly Val Val Leu Gly Gly Gln Val Arg Thr Phe
Gly Gly 530 535 540Lys Cys Tyr Asn Cys
Gly Gln Ile Gly His Leu Lys Lys Asn Cys Pro545 550
555 560Val Leu Asn Lys Gln Asn Ile Thr Ile Gln
Ala Thr Thr Thr Gly Arg 565 570
575Glu Pro Pro Asp Leu Cys Pro Arg Cys Lys Lys Gly Lys His Trp Ala
580 585 590Ser Gln Cys Arg Ser
Lys Phe Asp Lys Asn Gly Gln Pro Leu Ser Gly 595
600 605Asn Glu Gln Arg Gly Gln Pro Gln Ala Pro Gln Gln
Thr Gly Ala Phe 610 615 620Pro Ile Gln
Pro Phe Val Pro Gln Gly Phe Gln Gly Gln Gln Pro Pro625
630 635 640Leu Ser Gln Val Phe Gln Gly
Ile Ser Gln Leu Pro Gln Tyr Asn Asn 645
650 655Cys Pro Pro Pro Gln Ala Ala Val Gln Gln
660 665201710PRTArtificial SequenceDescription of
Artificial Sequence Synthetic polypeptide 20Met Ser Ser Glu Thr Gly
Pro Val Ala Val Asp Pro Thr Leu Arg Arg1 5
10 15Arg Ile Glu Pro His Glu Phe Glu Val Phe Phe Asp
Pro Arg Glu Leu 20 25 30Arg
Lys Glu Thr Cys Leu Leu Tyr Glu Ile Asn Trp Gly Gly Arg His 35
40 45Ser Ile Trp Arg His Thr Ser Gln Asn
Thr Asn Lys His Val Glu Val 50 55
60Asn Phe Ile Glu Lys Phe Thr Thr Glu Arg Tyr Phe Cys Pro Asn Thr65
70 75 80Arg Cys Ser Ile Thr
Trp Phe Leu Ser Trp Ser Pro Cys Gly Glu Cys 85
90 95Ser Arg Ala Ile Thr Glu Phe Leu Ser Arg Tyr
Pro His Val Thr Leu 100 105
110Phe Ile Tyr Ile Ala Arg Leu Tyr His His Ala Asp Pro Arg Asn Arg
115 120 125Gln Gly Leu Arg Asp Leu Ile
Ser Ser Gly Val Thr Ile Gln Ile Met 130 135
140Thr Glu Gln Glu Ser Gly Tyr Cys Trp Arg Asn Phe Val Asn Tyr
Ser145 150 155 160Pro Ser
Asn Glu Ala His Trp Pro Arg Tyr Pro His Leu Trp Val Arg
165 170 175Leu Tyr Val Leu Glu Leu Tyr
Cys Ile Ile Leu Gly Leu Pro Pro Cys 180 185
190Leu Asn Ile Leu Arg Arg Lys Gln Pro Gln Leu Thr Phe Phe
Thr Ile 195 200 205Ala Leu Gln Ser
Cys His Tyr Gln Arg Leu Pro Pro His Ile Leu Trp 210
215 220Ala Thr Gly Leu Lys Ser Gly Ser Glu Thr Pro Gly
Thr Ser Glu Ser225 230 235
240Ala Thr Pro Glu Ser Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly
245 250 255Thr Asn Ser Val Gly
Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro 260
265 270Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg
His Ser Ile Lys 275 280 285Lys Asn
Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 290
295 300Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg
Tyr Thr Arg Arg Lys305 310 315
320Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys
325 330 335Val Asp Asp Ser
Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 340
345 350Glu Asp Lys Lys His Glu Arg His Pro Ile Phe
Gly Asn Ile Val Asp 355 360 365Glu
Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys 370
375 380Lys Leu Val Asp Ser Thr Asp Lys Ala Asp
Leu Arg Leu Ile Tyr Leu385 390 395
400Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu
Gly 405 410 415Asp Leu Asn
Pro Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 420
425 430Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu
Asn Pro Ile Asn Ala Ser 435 440
445Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 450
455 460Arg Leu Glu Asn Leu Ile Ala Gln
Leu Pro Gly Glu Lys Lys Asn Gly465 470
475 480Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu
Thr Pro Asn Phe 485 490
495Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys
500 505 510Asp Thr Tyr Asp Asp Asp
Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp 515 520
525Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp
Ala Ile 530 535 540Leu Leu Ser Asp Ile
Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro545 550
555 560Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp
Glu His His Gln Asp Leu 565 570
575Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys
580 585 590Glu Ile Phe Phe Asp
Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 595
600 605Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile
Lys Pro Ile Leu 610 615 620Glu Lys Met
Asp Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu625
630 635 640Asp Leu Leu Arg Lys Gln Arg
Thr Phe Asp Asn Gly Ser Ile Pro His 645
650 655Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg
Arg Gln Glu Asp 660 665 670Phe
Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu 675
680 685Thr Phe Arg Ile Pro Tyr Tyr Val Gly
Pro Leu Ala Arg Gly Asn Ser 690 695
700Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp705
710 715 720Asn Phe Glu Glu
Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 725
730 735Glu Arg Met Thr Asn Phe Asp Lys Asn Leu
Pro Asn Glu Lys Val Leu 740 745
750Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu
755 760 765Thr Lys Val Lys Tyr Val Thr
Glu Gly Met Arg Lys Pro Ala Phe Leu 770 775
780Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr
Asn785 790 795 800Arg Lys
Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile
805 810 815Glu Cys Phe Asp Ser Val Glu
Ile Ser Gly Val Glu Asp Arg Phe Asn 820 825
830Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys
Asp Lys 835 840 845Asp Phe Leu Asp
Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 850
855 860Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile
Glu Glu Arg Leu865 870 875
880Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys
885 890 895Arg Arg Arg Tyr Thr
Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn 900
905 910Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu
Asp Phe Leu Lys 915 920 925Ser Asp
Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 930
935 940Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala
Gln Val Ser Gly Gln945 950 955
960Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala
965 970 975Ile Lys Lys Gly
Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val 980
985 990Lys Val Met Gly Arg His Lys Pro Glu Asn Ile
Val Ile Glu Met Ala 995 1000
1005Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu
1010 1015 1020Arg Met Lys Arg Ile Glu
Glu Gly Ile Lys Glu Leu Gly Ser Gln 1025 1030
1035Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn
Glu 1040 1045 1050Lys Leu Tyr Leu Tyr
Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val 1055 1060
1065Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp
Val Asp 1070 1075 1080His Ile Val Pro
Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn 1085
1090 1095Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly
Lys Ser Asp Asn 1100 1105 1110Val Pro
Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg 1115
1120 1125Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln
Arg Lys Phe Asp Asn 1130 1135 1140Leu
Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala 1145
1150 1155Gly Phe Ile Lys Arg Gln Leu Val Glu
Thr Arg Gln Ile Thr Lys 1160 1165
1170His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp
1175 1180 1185Glu Asn Asp Lys Leu Ile
Arg Glu Val Lys Val Ile Thr Leu Lys 1190 1195
1200Ser Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr
Lys 1205 1210 1215Val Arg Glu Ile Asn
Asn Tyr His His Ala His Asp Ala Tyr Leu 1220 1225
1230Asn Ala Val Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro
Lys Leu 1235 1240 1245Glu Ser Glu Phe
Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg 1250
1255 1260Lys Met Ile Ala Lys Ser Glu Gln Glu Ile Gly
Lys Ala Thr Ala 1265 1270 1275Lys Tyr
Phe Phe Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu 1280
1285 1290Ile Thr Leu Ala Asn Gly Glu Ile Arg Lys
Arg Pro Leu Ile Glu 1295 1300 1305Thr
Asn Gly Glu Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp 1310
1315 1320Phe Ala Thr Val Arg Lys Val Leu Ser
Met Pro Gln Val Asn Ile 1325 1330
1335Val Lys Lys Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser
1340 1345 1350Ile Leu Pro Lys Arg Asn
Ser Asp Lys Leu Ile Ala Arg Lys Lys 1355 1360
1365Asp Trp Asp Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr
Val 1370 1375 1380Ala Tyr Ser Val Leu
Val Val Ala Lys Val Glu Lys Gly Lys Ser 1385 1390
1395Lys Lys Leu Lys Ser Val Lys Glu Leu Leu Gly Ile Thr
Ile Met 1400 1405 1410Glu Arg Ser Ser
Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala 1415
1420 1425Lys Gly Tyr Lys Glu Val Lys Lys Asp Leu Ile
Ile Lys Leu Pro 1430 1435 1440Lys Tyr
Ser Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu 1445
1450 1455Ala Ser Ala Gly Glu Leu Gln Lys Gly Asn
Glu Leu Ala Leu Pro 1460 1465 1470Ser
Lys Tyr Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys 1475
1480 1485Leu Lys Gly Ser Pro Glu Asp Asn Glu
Gln Lys Gln Leu Phe Val 1490 1495
1500Glu Gln His Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser
1505 1510 1515Glu Phe Ser Lys Arg Val
Ile Leu Ala Asp Ala Asn Leu Asp Lys 1520 1525
1530Val Leu Ser Ala Tyr Asn Lys His Arg Asp Lys Pro Ile Arg
Glu 1535 1540 1545Gln Ala Glu Asn Ile
Ile His Leu Phe Thr Leu Thr Asn Leu Gly 1550 1555
1560Ala Pro Ala Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp
Arg Lys 1565 1570 1575Arg Tyr Thr Ser
Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His 1580
1585 1590Gln Ser Ile Thr Gly Leu Tyr Glu Thr Arg Ile
Asp Leu Ser Gln 1595 1600 1605Leu Gly
Gly Asp Ser Gly Gly Ser Thr Asn Leu Ser Asp Ile Ile 1610
1615 1620Glu Lys Glu Thr Gly Lys Gln Leu Val Ile
Gln Glu Ser Ile Leu 1625 1630 1635Met
Leu Pro Glu Glu Val Glu Glu Val Ile Gly Asn Lys Pro Glu 1640
1645 1650Ser Asp Ile Leu Val His Thr Ala Tyr
Asp Glu Ser Thr Asp Glu 1655 1660
1665Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr Lys Pro Trp
1670 1675 1680Ala Leu Val Ile Gln Asp
Ser Asn Gly Glu Asn Lys Ile Lys Met 1685 1690
1695Leu Ser Gly Gly Ser Pro Lys Lys Lys Arg Lys Val 1700
1705 1710211419PRTStreptococcus pyogenes
21Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val1
5 10 15Gly Trp Ala Val Ile Thr
Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 20 25
30Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys
Asn Leu Ile 35 40 45Gly Ala Leu
Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 50
55 60Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys
Asn Arg Ile Cys65 70 75
80Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser
85 90 95Phe Phe His Arg Leu Glu
Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 100
105 110His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp
Glu Val Ala Tyr 115 120 125His Glu
Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 130
135 140Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr
Leu Ala Leu Ala His145 150 155
160Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro
165 170 175Asp Asn Ser Asp
Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 180
185 190Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala
Ser Gly Val Asp Ala 195 200 205Lys
Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 210
215 220Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys
Asn Gly Leu Phe Gly Asn225 230 235
240Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn
Phe 245 250 255Asp Leu Ala
Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 260
265 270Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile
Gly Asp Gln Tyr Ala Asp 275 280
285Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 290
295 300Ile Leu Arg Val Asn Thr Glu Ile
Thr Lys Ala Pro Leu Ser Ala Ser305 310
315 320Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu
Thr Leu Leu Lys 325 330
335Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe
340 345 350Asp Gln Ser Lys Asn Gly
Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 355 360
365Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys
Met Asp 370 375 380Gly Thr Glu Glu Leu
Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg385 390
395 400Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile
Pro His Gln Ile His Leu 405 410
415Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe
420 425 430Leu Lys Asp Asn Arg
Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 435
440 445Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser
Arg Phe Ala Trp 450 455 460Met Thr Arg
Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu465
470 475 480Val Val Asp Lys Gly Ala Ser
Ala Gln Ser Phe Ile Glu Arg Met Thr 485
490 495Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu
Pro Lys His Ser 500 505 510Leu
Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 515
520 525Tyr Val Thr Glu Gly Met Arg Lys Pro
Ala Phe Leu Ser Gly Glu Gln 530 535
540Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr545
550 555 560Val Lys Gln Leu
Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 565
570 575Ser Val Glu Ile Ser Gly Val Glu Asp Arg
Phe Asn Ala Ser Leu Gly 580 585
590Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp
595 600 605Asn Glu Glu Asn Glu Asp Ile
Leu Glu Asp Ile Val Leu Thr Leu Thr 610 615
620Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr
Ala625 630 635 640His Leu
Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr
645 650 655Thr Gly Trp Gly Arg Leu Ser
Arg Lys Leu Ile Asn Gly Ile Arg Asp 660 665
670Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp
Gly Phe 675 680 685Ala Asn Arg Asn
Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 690
695 700Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln
Gly Asp Ser Leu705 710 715
720His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly
725 730 735Ile Leu Gln Thr Val
Lys Val Val Asp Glu Leu Val Lys Val Met Gly 740
745 750Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala
Arg Glu Asn Gln 755 760 765Thr Thr
Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 770
775 780Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile
Leu Lys Glu His Pro785 790 795
800Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu
805 810 815Gln Asn Gly Arg
Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 820
825 830Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro
Gln Ser Phe Leu Lys 835 840 845Asp
Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 850
855 860Gly Lys Ser Asp Asn Val Pro Ser Glu Glu
Val Val Lys Lys Met Lys865 870 875
880Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg
Lys 885 890 895Phe Asp Asn
Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 900
905 910Lys Ala Gly Phe Ile Lys Arg Gln Leu Val
Glu Thr Arg Gln Ile Thr 915 920
925Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 930
935 940Glu Asn Asp Lys Leu Ile Arg Glu
Val Lys Val Ile Thr Leu Lys Ser945 950
955 960Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe
Tyr Lys Val Arg 965 970
975Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val
980 985 990Val Gly Thr Ala Leu Ile
Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe 995 1000
1005Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys
Met Ile Ala 1010 1015 1020Lys Ser Glu
Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe Phe 1025
1030 1035Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu
Ile Thr Leu Ala 1040 1045 1050Asn Gly
Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly Glu 1055
1060 1065Thr Gly Glu Ile Val Trp Asp Lys Gly Arg
Asp Phe Ala Thr Val 1070 1075 1080Arg
Lys Val Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys Thr 1085
1090 1095Glu Val Gln Thr Gly Gly Phe Ser Lys
Glu Ser Ile Leu Pro Lys 1100 1105
1110Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp Pro
1115 1120 1125Lys Lys Tyr Gly Gly Phe
Asp Ser Pro Thr Val Ala Tyr Ser Val 1130 1135
1140Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu
Lys 1145 1150 1155Ser Val Lys Glu Leu
Leu Gly Ile Thr Ile Met Glu Arg Ser Ser 1160 1165
1170Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly
Tyr Lys 1175 1180 1185Glu Val Lys Lys
Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser Leu 1190
1195 1200Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu
Ala Ser Ala Gly 1205 1210 1215Glu Leu
Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr Val 1220
1225 1230Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu
Lys Leu Lys Gly Ser 1235 1240 1245Pro
Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His Lys 1250
1255 1260His Tyr Leu Asp Glu Ile Ile Glu Gln
Ile Ser Glu Phe Ser Lys 1265 1270
1275Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala
1280 1285 1290Tyr Asn Lys His Arg Asp
Lys Pro Ile Arg Glu Gln Ala Glu Asn 1295 1300
1305Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala
Ala 1310 1315 1320Phe Lys Tyr Phe Asp
Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser 1325 1330
1335Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser
Ile Thr 1340 1345 1350Gly Leu Tyr Glu
Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp 1355
1360 1365Gly Ser Gly Gly Gly Gly Ser Gly Lys Arg Thr
Ala Asp Gly Ser 1370 1375 1380Glu Phe
Glu Pro Lys Lys Lys Arg Lys Val Ser Ser Gly Gly Asp 1385
1390 1395Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp
His Asp Ile Asp Tyr 1400 1405 1410Lys
Asp Asp Asp Asp Lys 1415221053PRTStaphylococcus aureus 22Met Lys Arg
Asn Tyr Ile Leu Gly Leu Asp Ile Gly Ile Thr Ser Val1 5
10 15Gly Tyr Gly Ile Ile Asp Tyr Glu Thr
Arg Asp Val Ile Asp Ala Gly 20 25
30Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg
35 40 45Ser Lys Arg Gly Ala Arg Arg
Leu Lys Arg Arg Arg Arg His Arg Ile 50 55
60Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His65
70 75 80Ser Glu Leu Ser
Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 85
90 95Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser
Ala Ala Leu Leu His Leu 100 105
110Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr
115 120 125Gly Asn Glu Leu Ser Thr Lys
Glu Gln Ile Ser Arg Asn Ser Lys Ala 130 135
140Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys
Lys145 150 155 160Asp Gly
Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr
165 170 175Val Lys Glu Ala Lys Gln Leu
Leu Lys Val Gln Lys Ala Tyr His Gln 180 185
190Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu
Thr Arg 195 200 205Arg Thr Tyr Tyr
Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 210
215 220Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His
Cys Thr Tyr Phe225 230 235
240Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr
245 250 255Asn Ala Leu Asn Asp
Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 260
265 270Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile
Glu Asn Val Phe 275 280 285Lys Gln
Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 290
295 300Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val
Thr Ser Thr Gly Lys305 310 315
320Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr
325 330 335Ala Arg Lys Glu
Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 340
345 350Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp
Ile Gln Glu Glu Leu 355 360 365Thr
Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 370
375 380Asn Leu Lys Gly Tyr Thr Gly Thr His Asn
Leu Ser Leu Lys Ala Ile385 390 395
400Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile
Ala 405 410 415Ile Phe Asn
Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 420
425 430Gln Lys Glu Ile Pro Thr Thr Leu Val Asp
Asp Phe Ile Leu Ser Pro 435 440
445Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 450
455 460Ile Lys Lys Tyr Gly Leu Pro Asn
Asp Ile Ile Ile Glu Leu Ala Arg465 470
475 480Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn
Glu Met Gln Lys 485 490
495Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr
500 505 510Gly Lys Glu Asn Ala Lys
Tyr Leu Ile Glu Lys Ile Lys Leu His Asp 515 520
525Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro
Leu Glu 530 535 540Asp Leu Leu Asn Asn
Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro545 550
555 560Arg Ser Val Ser Phe Asp Asn Ser Phe Asn
Asn Lys Val Leu Val Lys 565 570
575Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu
580 585 590Ser Ser Ser Asp Ser
Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 595
600 605Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys
Thr Lys Lys Glu 610 615 620Tyr Leu Leu
Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp625
630 635 640Phe Ile Asn Arg Asn Leu Val
Asp Thr Arg Tyr Ala Thr Arg Gly Leu 645
650 655Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn
Leu Asp Val Lys 660 665 670Val
Lys Ser Ile Asn Gly Gly Phe Thr Ser Phe Leu Arg Arg Lys Trp 675
680 685Lys Phe Lys Lys Glu Arg Asn Lys Gly
Tyr Lys His His Ala Glu Asp 690 695
700Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys705
710 715 720Leu Asp Lys Ala
Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 725
730 735Gln Ala Glu Ser Met Pro Glu Ile Glu Thr
Glu Gln Glu Tyr Lys Glu 740 745
750Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp
755 760 765Tyr Lys Tyr Ser His Arg Val
Asp Lys Lys Pro Asn Arg Glu Leu Ile 770 775
780Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr
Leu785 790 795 800Ile Val
Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu
805 810 815Lys Lys Leu Ile Asn Lys Ser
Pro Glu Lys Leu Leu Met Tyr His His 820 825
830Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln
Tyr Gly 835 840 845Asp Glu Lys Asn
Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 850
855 860Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val
Ile Lys Lys Ile865 870 875
880Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp
885 890 895Tyr Pro Asn Ser Arg
Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 900
905 910Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys
Phe Val Thr Val 915 920 925Lys Asn
Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 930
935 940Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys
Ile Ser Asn Gln Ala945 950 955
960Glu Phe Ile Ala Ser Phe Tyr Asn Asn Asp Leu Ile Lys Ile Asn Gly
965 970 975Glu Leu Tyr Arg
Val Ile Gly Val Asn Asn Asp Leu Leu Asn Arg Ile 980
985 990Glu Val Asn Met Ile Asp Ile Thr Tyr Arg Glu
Tyr Leu Glu Asn Met 995 1000
1005Asn Asp Lys Arg Pro Pro Arg Ile Ile Lys Thr Ile Ala Ser Lys
1010 1015 1020Thr Gln Ser Ile Lys Lys
Tyr Ser Thr Asp Ile Leu Gly Asn Leu 1025 1030
1035Tyr Glu Val Lys Ser Lys Lys His Pro Gln Ile Ile Lys Lys
Gly 1040 1045
1050231307PRTAcidaminococcus sp. 23Met Thr Gln Phe Glu Gly Phe Thr Asn
Leu Tyr Gln Val Ser Lys Thr1 5 10
15Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu Lys His Ile
Gln 20 25 30Glu Gln Gly Phe
Ile Glu Glu Asp Lys Ala Arg Asn Asp His Tyr Lys 35
40 45Glu Leu Lys Pro Ile Ile Asp Arg Ile Tyr Lys Thr
Tyr Ala Asp Gln 50 55 60Cys Leu Gln
Leu Val Gln Leu Asp Trp Glu Asn Leu Ser Ala Ala Ile65 70
75 80Asp Ser Tyr Arg Lys Glu Lys Thr
Glu Glu Thr Arg Asn Ala Leu Ile 85 90
95Glu Glu Gln Ala Thr Tyr Arg Asn Ala Ile His Asp Tyr Phe
Ile Gly 100 105 110Arg Thr Asp
Asn Leu Thr Asp Ala Ile Asn Lys Arg His Ala Glu Ile 115
120 125Tyr Lys Gly Leu Phe Lys Ala Glu Leu Phe Asn
Gly Lys Val Leu Lys 130 135 140Gln Leu
Gly Thr Val Thr Thr Thr Glu His Glu Asn Ala Leu Leu Arg145
150 155 160Ser Phe Asp Lys Phe Thr Thr
Tyr Phe Ser Gly Phe Tyr Glu Asn Arg 165
170 175Lys Asn Val Phe Ser Ala Glu Asp Ile Ser Thr Ala
Ile Pro His Arg 180 185 190Ile
Val Gln Asp Asn Phe Pro Lys Phe Lys Glu Asn Cys His Ile Phe 195
200 205Thr Arg Leu Ile Thr Ala Val Pro Ser
Leu Arg Glu His Phe Glu Asn 210 215
220Val Lys Lys Ala Ile Gly Ile Phe Val Ser Thr Ser Ile Glu Glu Val225
230 235 240Phe Ser Phe Pro
Phe Tyr Asn Gln Leu Leu Thr Gln Thr Gln Ile Asp 245
250 255Leu Tyr Asn Gln Leu Leu Gly Gly Ile Ser
Arg Glu Ala Gly Thr Glu 260 265
270Lys Ile Lys Gly Leu Asn Glu Val Leu Asn Leu Ala Ile Gln Lys Asn
275 280 285Asp Glu Thr Ala His Ile Ile
Ala Ser Leu Pro His Arg Phe Ile Pro 290 295
300Leu Phe Lys Gln Ile Leu Ser Asp Arg Asn Thr Leu Ser Phe Ile
Leu305 310 315 320Glu Glu
Phe Lys Ser Asp Glu Glu Val Ile Gln Ser Phe Cys Lys Tyr
325 330 335Lys Thr Leu Leu Arg Asn Glu
Asn Val Leu Glu Thr Ala Glu Ala Leu 340 345
350Phe Asn Glu Leu Asn Ser Ile Asp Leu Thr His Ile Phe Ile
Ser His 355 360 365Lys Lys Leu Glu
Thr Ile Ser Ser Ala Leu Cys Asp His Trp Asp Thr 370
375 380Leu Arg Asn Ala Leu Tyr Glu Arg Arg Ile Ser Glu
Leu Thr Gly Lys385 390 395
400Ile Thr Lys Ser Ala Lys Glu Lys Val Gln Arg Ser Leu Lys His Glu
405 410 415Asp Ile Asn Leu Gln
Glu Ile Ile Ser Ala Ala Gly Lys Glu Leu Ser 420
425 430Glu Ala Phe Lys Gln Lys Thr Ser Glu Ile Leu Ser
His Ala His Ala 435 440 445Ala Leu
Asp Gln Pro Leu Pro Thr Thr Leu Lys Lys Gln Glu Glu Lys 450
455 460Glu Ile Leu Lys Ser Gln Leu Asp Ser Leu Leu
Gly Leu Tyr His Leu465 470 475
480Leu Asp Trp Phe Ala Val Asp Glu Ser Asn Glu Val Asp Pro Glu Phe
485 490 495Ser Ala Arg Leu
Thr Gly Ile Lys Leu Glu Met Glu Pro Ser Leu Ser 500
505 510Phe Tyr Asn Lys Ala Arg Asn Tyr Ala Thr Lys
Lys Pro Tyr Ser Val 515 520 525Glu
Lys Phe Lys Leu Asn Phe Gln Met Pro Thr Leu Ala Ser Gly Trp 530
535 540Asp Val Asn Lys Glu Lys Asn Asn Gly Ala
Ile Leu Phe Val Lys Asn545 550 555
560Gly Leu Tyr Tyr Leu Gly Ile Met Pro Lys Gln Lys Gly Arg Tyr
Lys 565 570 575Ala Leu Ser
Phe Glu Pro Thr Glu Lys Thr Ser Glu Gly Phe Asp Lys 580
585 590Met Tyr Tyr Asp Tyr Phe Pro Asp Ala Ala
Lys Met Ile Pro Lys Cys 595 600
605Ser Thr Gln Leu Lys Ala Val Thr Ala His Phe Gln Thr His Thr Thr 610
615 620Pro Ile Leu Leu Ser Asn Asn Phe
Ile Glu Pro Leu Glu Ile Thr Lys625 630
635 640Glu Ile Tyr Asp Leu Asn Asn Pro Glu Lys Glu Pro
Lys Lys Phe Gln 645 650
655Thr Ala Tyr Ala Lys Lys Thr Gly Asp Gln Lys Gly Tyr Arg Glu Ala
660 665 670Leu Cys Lys Trp Ile Asp
Phe Thr Arg Asp Phe Leu Ser Lys Tyr Thr 675 680
685Lys Thr Thr Ser Ile Asp Leu Ser Ser Leu Arg Pro Ser Ser
Gln Tyr 690 695 700Lys Asp Leu Gly Glu
Tyr Tyr Ala Glu Leu Asn Pro Leu Leu Tyr His705 710
715 720Ile Ser Phe Gln Arg Ile Ala Glu Lys Glu
Ile Met Asp Ala Val Glu 725 730
735Thr Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ala Lys
740 745 750Gly His His Gly Lys
Pro Asn Leu His Thr Leu Tyr Trp Thr Gly Leu 755
760 765Phe Ser Pro Glu Asn Leu Ala Lys Thr Ser Ile Lys
Leu Asn Gly Gln 770 775 780Ala Glu Leu
Phe Tyr Arg Pro Lys Ser Arg Met Lys Arg Met Ala His785
790 795 800Arg Leu Gly Glu Lys Met Leu
Asn Lys Lys Leu Lys Asp Gln Lys Thr 805
810 815Pro Ile Pro Asp Thr Leu Tyr Gln Glu Leu Tyr Asp
Tyr Val Asn His 820 825 830Arg
Leu Ser His Asp Leu Ser Asp Glu Ala Arg Ala Leu Leu Pro Asn 835
840 845Val Ile Thr Lys Glu Val Ser His Glu
Ile Ile Lys Asp Arg Arg Phe 850 855
860Thr Ser Asp Lys Phe Phe Phe His Val Pro Ile Thr Leu Asn Tyr Gln865
870 875 880Ala Ala Asn Ser
Pro Ser Lys Phe Asn Gln Arg Val Asn Ala Tyr Leu 885
890 895Lys Glu His Pro Glu Thr Pro Ile Ile Gly
Ile Asp Arg Gly Glu Arg 900 905
910Asn Leu Ile Tyr Ile Thr Val Ile Asp Ser Thr Gly Lys Ile Leu Glu
915 920 925Gln Arg Ser Leu Asn Thr Ile
Gln Gln Phe Asp Tyr Gln Lys Lys Leu 930 935
940Asp Asn Arg Glu Lys Glu Arg Val Ala Ala Arg Gln Ala Trp Ser
Val945 950 955 960Val Gly
Thr Ile Lys Asp Leu Lys Gln Gly Tyr Leu Ser Gln Val Ile
965 970 975His Glu Ile Val Asp Leu Met
Ile His Tyr Gln Ala Val Val Val Leu 980 985
990Glu Asn Leu Asn Phe Gly Phe Lys Ser Lys Arg Thr Gly Ile
Ala Glu 995 1000 1005Lys Ala Val
Tyr Gln Gln Phe Glu Lys Met Leu Ile Asp Lys Leu 1010
1015 1020Asn Cys Leu Val Leu Lys Asp Tyr Pro Ala Glu
Lys Val Gly Gly 1025 1030 1035Val Leu
Asn Pro Tyr Gln Leu Thr Asp Gln Phe Thr Ser Phe Ala 1040
1045 1050Lys Met Gly Thr Gln Ser Gly Phe Leu Phe
Tyr Val Pro Ala Pro 1055 1060 1065Tyr
Thr Ser Lys Ile Asp Pro Leu Thr Gly Phe Val Asp Pro Phe 1070
1075 1080Val Trp Lys Thr Ile Lys Asn His Glu
Ser Arg Lys His Phe Leu 1085 1090
1095Glu Gly Phe Asp Phe Leu His Tyr Asp Val Lys Thr Gly Asp Phe
1100 1105 1110Ile Leu His Phe Lys Met
Asn Arg Asn Leu Ser Phe Gln Arg Gly 1115 1120
1125Leu Pro Gly Phe Met Pro Ala Trp Asp Ile Val Phe Glu Lys
Asn 1130 1135 1140Glu Thr Gln Phe Asp
Ala Lys Gly Thr Pro Phe Ile Ala Gly Lys 1145 1150
1155Arg Ile Val Pro Val Ile Glu Asn His Arg Phe Thr Gly
Arg Tyr 1160 1165 1170Arg Asp Leu Tyr
Pro Ala Asn Glu Leu Ile Ala Leu Leu Glu Glu 1175
1180 1185Lys Gly Ile Val Phe Arg Asp Gly Ser Asn Ile
Leu Pro Lys Leu 1190 1195 1200Leu Glu
Asn Asp Asp Ser His Ala Ile Asp Thr Met Val Ala Leu 1205
1210 1215Ile Arg Ser Val Leu Gln Met Arg Asn Ser
Asn Ala Ala Thr Gly 1220 1225 1230Glu
Asp Tyr Ile Asn Ser Pro Val Arg Asp Leu Asn Gly Val Cys 1235
1240 1245Phe Asp Ser Arg Phe Gln Asn Pro Glu
Trp Pro Met Asp Ala Asp 1250 1255
1260Ala Asn Gly Ala Tyr His Ile Ala Leu Lys Gly Gln Leu Leu Leu
1265 1270 1275Asn His Leu Lys Glu Ser
Lys Asp Leu Lys Leu Gln Asn Gly Ile 1280 1285
1290Ser Asn Gln Asp Trp Leu Ala Tyr Ile Gln Glu Leu Arg Asn
1295 1300 130524175PRTHomo sapiens 24Met
Asp Ser Gly Arg Asp Phe Leu Thr Leu His Gly Leu Gln Asp Asp1
5 10 15Glu Asp Leu Gln Ala Leu Leu
Lys Gly Ser Gln Leu Leu Lys Val Lys 20 25
30Ser Ser Ser Trp Arg Arg Glu Arg Phe Tyr Lys Leu Gln Glu
Asp Cys 35 40 45Lys Thr Ile Trp
Gln Glu Ser Arg Lys Val Met Arg Thr Pro Glu Ser 50 55
60Gln Leu Phe Ser Ile Glu Asp Ile Gln Glu Val Arg Met
Gly His Arg65 70 75
80Thr Glu Gly Leu Glu Lys Phe Ala Arg Asp Val Pro Glu Asp Arg Cys
85 90 95Phe Ser Ile Val Phe Lys
Asp Gln Arg Asn Thr Leu Asp Leu Ile Ala 100
105 110Pro Ser Pro Ala Asp Ala Gln His Trp Val Leu Gly
Leu His Lys Ile 115 120 125Ile His
His Ser Gly Ser Met Asp Gln Arg Gln Lys Leu Gln His Trp 130
135 140Ile His Ser Cys Leu Arg Lys Ala Asp Lys Asn
Lys Asp Asn Lys Met145 150 155
160Ser Phe Lys Glu Leu Gln Asn Phe Leu Lys Glu Leu Asn Ile Gln
165 170 17525168PRTHomo sapiens
25Met Ser Asp Val Ala Ile Val Lys Glu Gly Trp Leu His Lys Arg Gly1
5 10 15Glu Tyr Ile Lys Thr Trp
Arg Pro Arg Tyr Phe Leu Leu Lys Asn Asp 20 25
30Gly Thr Phe Ile Gly Tyr Lys Glu Arg Pro Gln Asp Val
Asp Gln Arg 35 40 45Glu Ala Pro
Leu Asn Asn Phe Ser Val Ala Gln Cys Gln Leu Met Lys 50
55 60Thr Glu Arg Pro Arg Pro Asn Thr Phe Ile Ile Arg
Cys Leu Gln Trp65 70 75
80Thr Thr Val Ile Glu Arg Thr Phe His Val Glu Thr Pro Glu Glu Arg
85 90 95Glu Glu Trp Thr Thr Ala
Ile Gln Thr Val Ala Asp Gly Leu Lys Lys 100
105 110Gln Glu Glu Glu Glu Met Asp Phe Arg Ser Gly Ser
Pro Ser Asp Asn 115 120 125Ser Gly
Ala Glu Glu Met Glu Val Ser Leu Ala Lys Pro Lys His Arg 130
135 140Val Thr Met Asn Glu Phe Glu Tyr Leu Lys Leu
Leu Gly Lys Gly Thr145 150 155
160Phe Gly Lys Val Asp Pro Pro Val 1652692PRTHomo
sapiens 26Lys Met Gly Pro Val Asp Lys Arg Lys Gly Leu Phe Ala Arg Arg
Arg1 5 10 15Gln Leu Leu
Leu Thr Glu Gly Pro His Leu Tyr Tyr Val Asp Pro Val 20
25 30Asn Lys Val Leu Lys Gly Glu Ile Pro Trp
Ser Gln Glu Leu Arg Pro 35 40
45Glu Ala Lys Asn Phe Lys Thr Phe Phe Val His Thr Pro Asn Arg Thr 50
55 60Tyr Tyr Leu Met Asp Pro Ser Gly Asn
Ala His Lys Trp Cys Arg Lys65 70 75
80Ile Gln Glu Val Trp Arg Gln Arg Tyr Gln Ser His
85 9027229PRTHomo sapiens 27Met Ser Pro Val Lys Gly
Gly Thr Lys Cys Ile Lys Tyr Leu Leu Phe1 5
10 15Gly Phe Asn Phe Ile Phe Trp Leu Ala Gly Ile Ala
Val Leu Ala Ile 20 25 30Gly
Leu Trp Leu Arg Phe Asp Ser Gln Thr Lys Ser Ile Phe Glu Gln 35
40 45Glu Thr Asn Asn Asn Asn Ser Ser Phe
Tyr Thr Gly Val Tyr Ile Leu 50 55
60Ile Gly Ala Gly Ala Leu Met Met Leu Val Gly Phe Leu Gly Cys Cys65
70 75 80Gly Ala Val Gln Glu
Ser Gln Cys Met Leu Gly Leu Phe Phe Gly Phe 85
90 95Leu Leu Val Ile Phe Ala Ile Glu Ile Ala Ala
Ala Ile Trp Gly Tyr 100 105
110Ser His Lys Asp Glu Val Ile Lys Glu Val Gln Glu Phe Tyr Lys Asp
115 120 125Thr Tyr Asn Lys Leu Lys Thr
Lys Asp Glu Pro Gln Arg Glu Thr Leu 130 135
140Lys Ala Ile His Tyr Ala Leu Asn Cys Cys Gly Leu Ala Gly Gly
Val145 150 155 160Glu Gln
Phe Ile Ser Asp Ile Cys Pro Lys Lys Asp Val Leu Glu Thr
165 170 175Phe Thr Val Lys Ser Cys Pro
Asp Ala Ile Lys Glu Val Phe Asp Asn 180 185
190Lys Phe His Ile Ile Gly Ala Val Gly Ile Gly Ile Ala Val
Val Met 195 200 205Ile Phe Gly Met
Ile Phe Ser Met Ile Leu Cys Cys Ala Ile Arg Arg 210
215 220Asn Arg Glu Met Val22528238PRTHomo sapiens 28Met
Ala Val Glu Gly Gly Met Lys Cys Val Lys Phe Leu Leu Tyr Val1
5 10 15Leu Leu Leu Ala Phe Cys Ala
Cys Ala Val Gly Leu Ile Ala Val Gly 20 25
30Val Gly Ala Gln Leu Val Leu Ser Gln Thr Ile Ile Gln Gly
Ala Thr 35 40 45Pro Gly Ser Leu
Leu Pro Val Val Ile Ile Ala Val Gly Val Phe Leu 50 55
60Phe Leu Val Ala Phe Val Gly Cys Cys Gly Ala Cys Lys
Glu Asn Tyr65 70 75
80Cys Leu Met Ile Thr Phe Ala Ile Phe Leu Ser Leu Ile Met Leu Val
85 90 95Glu Val Ala Ala Ala Ile
Ala Gly Tyr Val Phe Arg Asp Lys Val Met 100
105 110Ser Glu Phe Asn Asn Asn Phe Arg Gln Gln Met Glu
Asn Tyr Pro Lys 115 120 125Asn Asn
His Thr Ala Ser Ile Leu Asp Arg Met Gln Ala Asp Phe Lys 130
135 140Cys Cys Gly Ala Ala Asn Tyr Thr Asp Trp Glu
Lys Ile Pro Ser Met145 150 155
160Ser Lys Asn Arg Val Pro Asp Ser Cys Cys Ile Asn Val Thr Val Gly
165 170 175Cys Gly Ile Asn
Phe Asn Glu Lys Ala Ile His Lys Glu Gly Cys Val 180
185 190Glu Lys Ile Gly Gly Trp Leu Arg Lys Asn Val
Leu Val Val Ala Ala 195 200 205Ala
Ala Leu Gly Ile Ala Phe Val Glu Val Leu Gly Ile Val Phe Ala 210
215 220Cys Cys Leu Val Lys Ser Ile Arg Ser Gly
Tyr Glu Val Met225 230 23529236PRTHomo
sapiens 29Met Gly Val Glu Gly Cys Thr Lys Cys Ile Lys Tyr Leu Leu Phe
Val1 5 10 15Phe Asn Phe
Val Phe Trp Leu Ala Gly Gly Val Ile Leu Gly Val Ala 20
25 30Leu Trp Leu Arg His Asp Pro Gln Thr Thr
Asn Leu Leu Tyr Leu Glu 35 40
45Leu Gly Asp Lys Pro Ala Pro Asn Thr Phe Tyr Val Gly Ile Tyr Ile 50
55 60Leu Ile Ala Val Gly Ala Val Met Met
Phe Val Gly Phe Leu Gly Cys65 70 75
80Tyr Gly Ala Ile Gln Glu Ser Gln Cys Leu Leu Gly Thr Phe
Phe Thr 85 90 95Cys Leu
Val Ile Leu Phe Ala Cys Glu Val Ala Ala Gly Ile Trp Gly 100
105 110Phe Val Asn Lys Asp Gln Ile Ala Lys
Asp Val Lys Gln Phe Tyr Asp 115 120
125Gln Ala Leu Gln Gln Ala Val Val Asp Asp Asp Ala Asn Asn Ala Lys
130 135 140Ala Val Val Lys Thr Phe His
Glu Thr Leu Asp Cys Cys Gly Ser Ser145 150
155 160Thr Leu Thr Ala Leu Thr Thr Ser Val Leu Lys Asn
Asn Leu Cys Pro 165 170
175Ser Gly Ser Asn Ile Ile Ser Asn Leu Phe Lys Glu Asp Cys His Gln
180 185 190Lys Ile Asp Asp Leu Phe
Ser Gly Lys Leu Tyr Leu Ile Gly Ile Ala 195 200
205Ala Ile Val Val Ala Val Ile Met Ile Phe Glu Met Ile Leu
Ser Met 210 215 220Val Leu Cys Cys Gly
Ile Arg Asn Ser Ser Val Tyr225 230
2353010PRTHomo sapiens 30Glu Val Thr Glu Leu Thr Arg Glu Gly Glu1
5 103121PRTHomo sapiens 31Gly Asn Tyr Thr Cys Glu
Val Thr Glu Leu Thr Arg Glu Gly Glu Thr1 5
10 15Ile Ile Glu Leu Lys 2032293PRTHomo
sapiens 32Met Trp Pro Leu Val Ala Ala Leu Leu Leu Gly Ser Ala Cys Cys
Gly1 5 10 15Ser Ala Gln
Leu Leu Phe Asn Lys Thr Lys Ser Val Glu Phe Thr Phe 20
25 30Cys Asn Asp Thr Val Val Ile Pro Cys Phe
Val Thr Asn Met Glu Ala 35 40
45Gln Asn Thr Thr Glu Val Tyr Val Lys Trp Lys Phe Lys Gly Arg Asp 50
55 60Ile Tyr Thr Phe Asp Gly Ala Leu Asn
Lys Ser Thr Val Pro Thr Asp65 70 75
80Phe Ser Ser Ala Lys Ile Glu Val Ser Gln Leu Leu Lys Gly
Asp Ala 85 90 95Ser Leu
Lys Met Asp Lys Ser Asp Ala Val Ser His Thr Gly Asn Tyr 100
105 110Thr Cys Glu Val Thr Glu Leu Thr Arg
Glu Gly Glu Thr Ile Ile Glu 115 120
125Leu Lys Tyr Arg Val Val Ser Trp Phe Ser Pro Asn Glu Asn Ile Leu
130 135 140Ile Val Ile Phe Pro Ile Phe
Ala Ile Leu Leu Phe Trp Gly Gln Phe145 150
155 160Gly Ile Lys Thr Leu Lys Tyr Arg Ser Gly Gly Met
Asp Glu Lys Thr 165 170
175Ile Ala Leu Leu Val Ala Gly Leu Val Ile Thr Val Ile Val Ile Val
180 185 190Gly Ala Ile Leu Phe Val
Pro Gly Glu Tyr Ser Leu Lys Asn Ala Thr 195 200
205Gly Leu Gly Leu Ile Val Thr Ser Thr Gly Ile Leu Ile Leu
Leu His 210 215 220Tyr Tyr Val Phe Ser
Thr Ala Ile Gly Leu Thr Ser Phe Val Ile Ala225 230
235 240Ile Leu Val Ile Gln Val Ile Ala Tyr Ile
Leu Ala Val Val Gly Leu 245 250
255Ser Leu Cys Ile Ala Ala Cys Ile Pro Met His Gly Pro Leu Leu Ile
260 265 270Ser Gly Leu Ser Ile
Leu Ala Leu Ala Gln Leu Leu Gly Leu Val Tyr 275
280 285Met Lys Phe Val Glu 29033110PRTArtificial
SequenceDescription of Artificial Sequence Synthetic polypeptide
33Phe Lys Cys Glu His Cys Arg Ile Leu Phe Leu Asp His Val Met Phe1
5 10 15Thr Ile His Met Gly Cys
His Gly Phe Arg Asp Pro Phe Lys Cys Asn 20 25
30Met Cys Gly Glu Lys Cys Asp Gly Pro Val Gly Leu Phe
Val His Met 35 40 45Ala Arg Asn
Ala His Gly Glu Lys Pro Phe Tyr Cys Glu His Cys Glu 50
55 60Ile Thr Phe Arg Asp Val Val Met Tyr Ser Leu His
Lys Gly Tyr His65 70 75
80Gly Phe Arg Asp Pro Phe Glu Cys Asn Ile Cys Gly Tyr His Ser Gln
85 90 95Asp Arg Tyr Glu Phe Ser
Ser His Ile Val Arg Gly Glu His 100 105
11034110PRTArtificial SequenceDescription of Artificial Sequence
Synthetic polypeptide 34His His Cys Gln His Cys Asp Met Tyr Phe Ala
Asp Asn Ile Leu Tyr1 5 10
15Thr Ile His Met Gly Cys His Ser Cys Asp Asp Val Phe Lys Cys Asn
20 25 30Met Cys Gly Glu Lys Cys Asp
Gly Pro Val Gly Leu Phe Val His Met 35 40
45Ala Arg Asn Ala His Gly Glu Lys Pro Thr Lys Cys Val His Cys
Gly 50 55 60Ile Val Phe Leu Asp Glu
Val Met Tyr Ala Leu His Met Ser Cys His65 70
75 80Gly Phe Arg Asp Pro Phe Glu Cys Asn Ile Cys
Gly Tyr His Ser Gln 85 90
95Asp Arg Tyr Glu Phe Ser Ser His Ile Val Arg Gly Glu His 100
105 11035110PRTArtificial
SequenceDescription of Artificial Sequence Synthetic polypeptide
35Met Gly Arg Gly Val Gln Val Glu Thr Ile Ser Pro Gly Asp Gly Arg1
5 10 15Thr Phe Pro Lys Arg Gly
Gln Thr Cys Val Val His Tyr Thr Gly Met 20 25
30Leu Glu Asp Gly Lys Lys Phe Asp Ser Ser Arg Asp Arg
Asn Lys Pro 35 40 45Phe Lys Phe
Met Leu Gly Lys Gln Glu Val Ile Arg Gly Trp Glu Glu 50
55 60Gly Val Ala Gln Met Ser Val Gly Gln Arg Ala Lys
Leu Thr Ile Ser65 70 75
80Pro Asp Tyr Ala Tyr Gly Ala Thr Gly His Pro Gly Ile Ile Pro Pro
85 90 95His Ala Thr Leu Val Phe
Asp Val Glu Leu Leu Lys Leu Glu 100 105
11036111PRTArtificial SequenceDescription of Artificial Sequence
Synthetic polypeptide 36Met Ala Ser Arg Gly Val Gln Val Glu Thr Ile
Ser Pro Gly Asp Gly1 5 10
15Arg Thr Phe Pro Lys Arg Gly Gln Thr Cys Val Val His Tyr Thr Gly
20 25 30Met Leu Glu Asp Gly Lys Lys
Val Asp Ser Ser Arg Asp Arg Asn Lys 35 40
45Pro Phe Lys Phe Met Leu Gly Lys Gln Glu Val Ile Arg Gly Trp
Glu 50 55 60Glu Gly Val Ala Gln Met
Ser Val Gly Gln Arg Ala Lys Leu Thr Ile65 70
75 80Ser Pro Asp Tyr Ala Tyr Gly Ala Thr Gly His
Pro Gly Ile Ile Pro 85 90
95Pro His Ala Thr Leu Val Phe Asp Val Glu Leu Leu Lys Leu Glu
100 105 1103797PRTArtificial
SequenceDescription of Artificial Sequence Synthetic polypeptide
37Met Gly Ser Arg Ile Leu Trp His Glu Met Trp His Glu Gly Leu Glu1
5 10 15Glu Ala Ser Arg Leu Tyr
Phe Gly Glu Arg Asn Val Lys Gly Met Phe 20 25
30Glu Val Leu Glu Pro Leu His Ala Met Met Glu Arg Gly
Pro Gln Thr 35 40 45Leu Lys Glu
Thr Ser Phe Asn Gln Ala Tyr Gly Arg Asp Leu Met Glu 50
55 60Ala Gln Glu Trp Cys Arg Lys Tyr Met Lys Ser Gly
Asn Val Lys Asp65 70 75
80Leu Leu Gln Ala Trp Asp Leu Tyr Tyr His Val Phe Arg Arg Ile Ser
85 90 95Lys38108PRTArtificial
SequenceDescription of Artificial Sequence Synthetic polypeptide
38Met Gly Val Gln Val Glu Thr Ile Ser Pro Gly Asp Gly Arg Thr Phe1
5 10 15Pro Lys Arg Gly Gln Thr
Cys Val Val His Tyr Thr Gly Met Leu Glu 20 25
30Asp Gly Lys Lys Phe Asp Ser Ser Arg Asp Arg Asn Lys
Pro Phe Lys 35 40 45Phe Met Leu
Gly Lys Gln Glu Val Ile Arg Gly Trp Glu Glu Gly Val 50
55 60Ala Gln Met Ser Val Gly Gln Arg Ala Lys Leu Thr
Ile Ser Pro Asp65 70 75
80Tyr Ala Tyr Gly Ala Thr Gly His Pro Gly Ile Ile Pro Pro His Ala
85 90 95Thr Leu Val Phe Asp Val
Glu Leu Leu Lys Leu Glu 100
1053993PRTArtificial SequenceDescription of Artificial Sequence Synthetic
polypeptide 39Gln Gly Met Leu Glu Met Trp His Glu Gly Leu Glu Glu
Ala Ser Arg1 5 10 15Leu
Tyr Phe Gly Glu Arg Asn Val Lys Gly Met Phe Glu Val Leu Glu 20
25 30Pro Leu His Ala Met Met Glu Arg
Gly Pro Gln Thr Leu Lys Glu Thr 35 40
45Ser Phe Asn Gln Ala Tyr Gly Arg Asp Leu Met Glu Ala Gln Glu Trp
50 55 60Cys Arg Lys Tyr Met Lys Ser Gly
Asn Val Lys Asp Leu Leu Gln Ala65 70 75
80Trp Asp Leu Tyr Tyr His Val Phe Arg Arg Ile Ser Lys
85 9040277PRTArtificial SequenceDescription
of Artificial Sequence Synthetic polypeptide 40Met Gly Pro Asp Ile
Val Met Thr Gln Ser Pro Ser Ser Leu Ser Ala1 5
10 15Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg
Ser Ser Thr Gly Ala 20 25
30Val Thr Thr Ser Asn Tyr Ala Ser Trp Val Gln Glu Lys Pro Gly Lys
35 40 45Leu Phe Lys Gly Leu Ile Gly Gly
Thr Asn Asn Arg Ala Pro Gly Val 50 55
60Pro Ser Arg Phe Ser Gly Ser Leu Ile Gly Asp Lys Ala Thr Leu Thr65
70 75 80Ile Ser Ser Leu Gln
Pro Glu Asp Phe Ala Thr Tyr Phe Cys Ala Leu 85
90 95Trp Tyr Ser Asn His Trp Val Phe Gly Gln Gly
Thr Lys Val Glu Leu 100 105
110Lys Arg Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
115 120 125Ser Ser Gly Gly Gly Ser Glu
Val Lys Leu Leu Glu Ser Gly Gly Gly 130 135
140Leu Val Gln Pro Gly Gly Ser Leu Lys Leu Ser Cys Ala Val Ser
Gly145 150 155 160Phe Ser
Leu Thr Asp Tyr Gly Val Asn Trp Val Arg Gln Ala Pro Gly
165 170 175Arg Gly Leu Glu Trp Ile Gly
Val Ile Trp Gly Asp Gly Ile Thr Asp 180 185
190Tyr Asn Ser Ala Leu Lys Asp Arg Phe Ile Ile Ser Lys Asp
Asn Gly 195 200 205Lys Asn Thr Val
Tyr Leu Gln Met Ser Lys Val Arg Ser Asp Asp Thr 210
215 220Ala Leu Tyr Tyr Cys Val Thr Gly Leu Phe Asp Tyr
Trp Gly Gln Gly225 230 235
240Thr Leu Val Thr Val Ser Ser Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
245 250 255Gly Gly Gly Gly Gly
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 260
265 270Gly Gly Gly Gly Ser 27541237PRTArtificial
SequenceDescription of Artificial Sequence Synthetic polypeptide
41Glu Glu Leu Leu Ser Lys Asn Tyr His Leu Glu Asn Glu Val Ala Arg1
5 10 15Leu Lys Lys Gly Ser Gly
Ser Gly Glu Glu Leu Leu Ser Lys Asn Tyr 20 25
30His Leu Glu Asn Glu Val Ala Arg Leu Lys Lys Gly Ser
Gly Ser Gly 35 40 45Glu Glu Leu
Leu Ser Lys Asn Tyr His Leu Glu Asn Glu Val Ala Arg 50
55 60Leu Lys Lys Gly Ser Gly Ser Gly Glu Glu Leu Leu
Ser Lys Asn Tyr65 70 75
80His Leu Glu Asn Glu Val Ala Arg Leu Lys Lys Gly Ser Gly Ser Gly
85 90 95Glu Glu Leu Leu Ser Lys
Asn Tyr His Leu Glu Asn Glu Val Ala Arg 100
105 110Leu Lys Lys Gly Ser Gly Ser Gly Glu Glu Leu Leu
Ser Lys Asn Tyr 115 120 125His Leu
Glu Asn Glu Val Ala Arg Leu Lys Lys Gly Ser Gly Ser Gly 130
135 140Glu Glu Leu Leu Ser Lys Asn Tyr His Leu Glu
Asn Glu Val Ala Arg145 150 155
160Leu Lys Lys Gly Ser Gly Ser Gly Glu Glu Leu Leu Ser Lys Asn Tyr
165 170 175His Leu Glu Asn
Glu Val Ala Arg Leu Lys Lys Gly Ser Gly Ser Gly 180
185 190Glu Glu Leu Leu Ser Lys Asn Tyr His Leu Glu
Asn Glu Val Ala Arg 195 200 205Leu
Lys Lys Gly Ser Gly Ser Gly Glu Glu Leu Leu Ser Lys Asn Tyr 210
215 220His Leu Glu Asn Glu Val Ala Arg Leu Lys
Lys Gly Ser225 230 23542573PRTArtificial
SequenceDescription of Artificial Sequence Synthetic polypeptide
42Glu Glu Leu Leu Ser Lys Asn Tyr His Leu Glu Asn Glu Val Ala Arg1
5 10 15Leu Lys Lys Gly Ser Gly
Ser Gly Glu Glu Leu Leu Ser Lys Asn Tyr 20 25
30His Leu Glu Asn Glu Val Ala Arg Leu Lys Lys Gly Ser
Gly Ser Gly 35 40 45Glu Glu Leu
Leu Ser Lys Asn Tyr His Leu Glu Asn Glu Val Ala Arg 50
55 60Leu Lys Lys Gly Ser Gly Ser Gly Glu Glu Leu Leu
Ser Lys Asn Tyr65 70 75
80His Leu Glu Asn Glu Val Ala Arg Leu Lys Lys Gly Ser Gly Ser Gly
85 90 95Glu Glu Leu Leu Ser Lys
Asn Tyr His Leu Glu Asn Glu Val Ala Arg 100
105 110Leu Lys Lys Gly Ser Gly Ser Gly Glu Glu Leu Leu
Ser Lys Asn Tyr 115 120 125His Leu
Glu Asn Glu Val Ala Arg Leu Lys Lys Gly Ser Gly Ser Gly 130
135 140Glu Glu Leu Leu Ser Lys Asn Tyr His Leu Glu
Asn Glu Val Ala Arg145 150 155
160Leu Lys Lys Gly Ser Gly Ser Gly Glu Glu Leu Leu Ser Lys Asn Tyr
165 170 175His Leu Glu Asn
Glu Val Ala Arg Leu Lys Lys Gly Ser Gly Ser Gly 180
185 190Glu Glu Leu Leu Ser Lys Asn Tyr His Leu Glu
Asn Glu Val Ala Arg 195 200 205Leu
Lys Lys Gly Ser Gly Ser Gly Glu Glu Leu Leu Ser Lys Asn Tyr 210
215 220His Leu Glu Asn Glu Val Ala Arg Leu Lys
Lys Gly Ser Gly Ser Gly225 230 235
240Glu Glu Leu Leu Ser Lys Asn Tyr His Leu Glu Asn Glu Val Ala
Arg 245 250 255Leu Lys Lys
Gly Ser Gly Ser Gly Glu Glu Leu Leu Ser Lys Asn Tyr 260
265 270His Leu Glu Asn Glu Val Ala Arg Leu Lys
Lys Gly Ser Gly Ser Gly 275 280
285Glu Glu Leu Leu Ser Lys Asn Tyr His Leu Glu Asn Glu Val Ala Arg 290
295 300Leu Lys Lys Gly Ser Gly Ser Gly
Glu Glu Leu Leu Ser Lys Asn Tyr305 310
315 320His Leu Glu Asn Glu Val Ala Arg Leu Lys Lys Gly
Ser Gly Ser Gly 325 330
335Glu Glu Leu Leu Ser Lys Asn Tyr His Leu Glu Asn Glu Val Ala Arg
340 345 350Leu Lys Lys Gly Ser Gly
Ser Gly Glu Glu Leu Leu Ser Lys Asn Tyr 355 360
365His Leu Glu Asn Glu Val Ala Arg Leu Lys Lys Gly Ser Gly
Ser Gly 370 375 380Glu Glu Leu Leu Ser
Lys Asn Tyr His Leu Glu Asn Glu Val Ala Arg385 390
395 400Leu Lys Lys Gly Ser Gly Ser Gly Glu Glu
Leu Leu Ser Lys Asn Tyr 405 410
415His Leu Glu Asn Glu Val Ala Arg Leu Lys Lys Gly Ser Gly Ser Gly
420 425 430Glu Glu Leu Leu Ser
Lys Asn Tyr His Leu Glu Asn Glu Val Ala Arg 435
440 445Leu Lys Lys Gly Ser Gly Ser Gly Glu Glu Leu Leu
Ser Lys Asn Tyr 450 455 460His Leu Glu
Asn Glu Val Ala Arg Leu Lys Lys Gly Ser Gly Ser Gly465
470 475 480Glu Glu Leu Leu Ser Lys Asn
Tyr His Leu Glu Asn Glu Val Ala Arg 485
490 495Leu Lys Lys Gly Ser Gly Ser Gly Glu Glu Leu Leu
Ser Lys Asn Tyr 500 505 510His
Leu Glu Asn Glu Val Ala Arg Leu Lys Lys Gly Ser Gly Ser Gly 515
520 525Glu Glu Leu Leu Ser Lys Asp Tyr His
Leu Glu Asn Glu Val Ala Arg 530 535
540Leu Lys Lys Gly Ser Gly Ser Gly Glu Glu Leu Leu Ser Lys Asn Tyr545
550 555 560His Leu Glu Asn
Glu Val Ala Arg Leu Lys Lys Gly Ser 565
57043114PRTArtificial SequenceDescription of Artificial Sequence
Synthetic polypeptide 43Val Gln Leu Val Glu Ser Gly Gly Ala Leu Val
Gln Pro Gly Gly Ser1 5 10
15Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Pro Val Asn Arg Tyr Ser
20 25 30Met Arg Trp Tyr Arg Gln Ala
Pro Gly Lys Glu Arg Glu Trp Val Ala 35 40
45Gly Met Ser Ser Ala Gly Asp Arg Ser Ser Tyr Glu Asp Ser Val
Lys 50 55 60Gly Arg Phe Thr Ile Ser
Arg Asp Asp Ala Arg Asn Thr Val Tyr Leu65 70
75 80Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala
Val Tyr Tyr Ser Asn 85 90
95Val Asn Val Gly Phe Glu Tyr Trp Gly Gln Gly Thr Gln Val Thr Val
100 105 110Ser Ser44102PRTNostoc
punctiforme 44Cys Leu Ser Tyr Glu Thr Glu Ile Leu Thr Val Glu Tyr Gly Leu
Leu1 5 10 15Pro Ile Gly
Lys Ile Val Glu Lys Arg Ile Glu Cys Thr Val Tyr Ser 20
25 30Val Asp Asn Asn Gly Asn Ile Tyr Thr Gln
Pro Val Ala Gln Trp His 35 40
45Asp Arg Gly Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu Asp Gly Ser 50
55 60Leu Ile Arg Ala Thr Lys Asp His Lys
Phe Met Thr Val Asp Gly Gln65 70 75
80Met Leu Pro Ile Asp Glu Ile Phe Glu Arg Glu Leu Asp Leu
Met Arg 85 90 95Val Asp
Asn Leu Pro Asn 1004539PRTNostoc punctiforme 45Met Ile Lys Ile
Ala Thr Arg Lys Tyr Leu Gly Lys Gln Asn Val Tyr1 5
10 15Asp Ile Gly Val Glu Arg Asp His Asn Phe
Ala Leu Lys Asn Gly Phe 20 25
30Ile Ala Ser Asn Cys Phe Asn 3546101PRTArtificial
SequenceDescription of Artificial Sequence Synthetic polypeptide
46Cys Leu Ser Tyr Asp Thr Glu Ile Leu Thr Val Glu Tyr Gly Phe Leu1
5 10 15Pro Ile Gly Lys Ile Val
Glu Glu Arg Ile Glu Cys Thr Val Tyr Thr 20 25
30Val Asp Lys Asn Gly Phe Val Tyr Thr Gln Pro Ile Ala
Gln Trp His 35 40 45Asn Arg Gly
Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu Asp Gly Ser 50
55 60Ile Ile Arg Ala Thr Lys Asp His Lys Phe Met Thr
Thr Asp Gly Gln65 70 75
80Met Leu Pro Ile Asp Glu Ile Phe Glu Arg Gly Leu Asp Leu Lys Gln
85 90 95Val Asp Gly Leu Pro
1004736PRTArtificial SequenceDescription of Artificial Sequence
Synthetic polypeptide 47Met Val Lys Ile Ile Ser Arg Lys Ser Leu Gly
Thr Gln Asn Val Tyr1 5 10
15Asp Ile Gly Val Glu Lys Asp His Asn Phe Leu Leu Lys Asn Gly Leu
20 25 30Val Ala Ser Asn
3548184PRTSaccharomyces cerevisiae 48Cys Phe Ala Lys Gly Thr Asn Val Leu
Met Ala Asp Gly Ser Ile Glu1 5 10
15Cys Ile Glu Asn Ile Glu Val Gly Asn Lys Val Met Gly Lys Asp
Gly 20 25 30Arg Pro Arg Glu
Val Ile Lys Leu Pro Arg Gly Arg Glu Thr Met Tyr 35
40 45Ser Val Val Gln Lys Ser Gln His Arg Ala His Lys
Ser Asp Ser Ser 50 55 60Arg Glu Val
Pro Glu Leu Leu Lys Phe Thr Cys Asn Ala Thr His Glu65 70
75 80Leu Val Val Arg Thr Pro Arg Ser
Val Arg Arg Leu Ser Arg Thr Ile 85 90
95Lys Gly Val Glu Tyr Phe Glu Val Ile Thr Phe Glu Met Gly
Gln Lys 100 105 110Lys Ala Pro
Asp Gly Arg Ile Val Glu Leu Val Lys Glu Val Ser Lys 115
120 125Ser Tyr Pro Ile Ser Glu Gly Pro Glu Arg Ala
Asn Glu Leu Val Glu 130 135 140Ser Tyr
Arg Lys Ala Ser Asn Lys Ala Tyr Phe Glu Trp Thr Ile Glu145
150 155 160Ala Arg Asp Leu Ser Leu Leu
Gly Ser His Val Arg Lys Ala Thr Tyr 165
170 175Gln Thr Tyr Ala Pro Ile Leu Tyr
1804965PRTSaccharomyces cerevisiae 49Val Leu Leu Asn Val Leu Ser Lys Cys
Ala Gly Ser Lys Lys Phe Arg1 5 10
15Pro Ala Pro Ala Ala Ala Phe Ala Arg Glu Cys Arg Gly Phe Tyr
Phe 20 25 30Glu Leu Gln Glu
Leu Lys Glu Asp Asp Tyr Tyr Gly Ile Thr Leu Ser 35
40 45Asp Asp Ser Asp His Gln Phe Leu Leu Ala Asn Gln
Val Val Val His 50 55
60Asn6550123PRTSynechocystis sp. 50Cys Leu Ser Phe Gly Thr Glu Ile Leu
Thr Val Glu Tyr Gly Pro Leu1 5 10
15Pro Ile Gly Lys Ile Val Ser Glu Glu Ile Asn Cys Ser Val Tyr
Ser 20 25 30Val Asp Pro Glu
Gly Arg Val Tyr Thr Gln Ala Ile Ala Gln Trp His 35
40 45Asp Arg Gly Glu Gln Glu Val Leu Glu Tyr Glu Leu
Glu Asp Gly Ser 50 55 60Val Ile Arg
Ala Thr Ser Asp His Arg Phe Leu Thr Thr Asp Tyr Gln65 70
75 80Leu Leu Ala Ile Glu Glu Ile Phe
Ala Arg Gln Leu Asp Leu Leu Thr 85 90
95Leu Glu Asn Ile Lys Gln Thr Glu Glu Ala Leu Asp Asn His
Arg Leu 100 105 110Pro Phe Pro
Leu Leu Asp Ala Gly Thr Ile Lys 115
1205136PRTSynechocystis sp. 51Met Val Lys Val Ile Gly Arg Arg Ser Leu Gly
Val Gln Arg Ile Phe1 5 10
15Asp Ile Gly Leu Pro Gln Asp His Asn Phe Leu Leu Ala Asn Gly Ala
20 25 30Ile Ala Ala Asn
355214PRTArtificial SequenceDescription of Artificial Sequence Synthetic
peptide 52Val Pro Thr Ile Val Met Val Asp Ala Tyr Lys Arg Tyr Lys1
5 1053119PRTArtificial SequenceDescription
of Artificial Sequence Synthetic polypeptide 53Met Val Thr Thr Leu
Ser Gly Leu Ser Gly Glu Gln Gly Pro Ser Gly1 5
10 15Asp Met Thr Thr Glu Glu Asp Ser Ala Thr His
Ile Lys Phe Ser Lys 20 25
30Arg Asp Glu Asp Gly Arg Glu Leu Ala Gly Ala Thr Met Glu Leu Arg
35 40 45Asp Ser Ser Gly Lys Thr Ile Ser
Thr Trp Ile Ser Asp Gly His Val 50 55
60Lys Asp Phe Tyr Leu Tyr Pro Gly Lys Tyr Thr Phe Val Glu Thr Ala65
70 75 80Ala Pro Asp Gly Tyr
Glu Val Ala Thr Ala Ile Thr Phe Thr Val Asn 85
90 95Glu Gln Gly Gln Val Thr Val Asn Gly Glu Ala
Thr Lys Gly Asp Ala 100 105
110His Thr Gly Ser Ser Gly Ser 11554130PRTEnterobacteria phage MS2
54Met Ala Ser Asn Phe Thr Gln Phe Val Leu Val Asp Asn Gly Gly Thr1
5 10 15Gly Asp Val Thr Val Ala
Pro Ser Asn Phe Ala Asn Gly Val Ala Glu 20 25
30Trp Ile Ser Ser Asn Ser Arg Ser Gln Ala Tyr Lys Val
Thr Cys Ser 35 40 45Val Arg Gln
Ser Ser Ala Gln Asn Arg Lys Tyr Thr Ile Lys Val Glu 50
55 60Val Pro Lys Val Ala Thr Gln Thr Val Gly Gly Val
Glu Leu Pro Val65 70 75
80Ala Ala Trp Arg Ser Tyr Leu Asn Met Glu Leu Thr Ile Pro Ile Phe
85 90 95Ala Thr Asn Ser Asp Cys
Glu Leu Ile Val Lys Ala Met Gln Gly Leu 100
105 110Leu Lys Asp Gly Asn Pro Ile Pro Ser Ala Ile Ala
Ala Asn Ser Gly 115 120 125Ile Tyr
13055130PRTEnterobacteria phage MS2 55Met Ala Ser Asn Phe Thr Gln Phe
Val Leu Val Asp Asn Gly Gly Thr1 5 10
15Gly Asp Val Thr Val Ala Pro Ser Asn Phe Ala Asn Gly Val
Ala Glu 20 25 30Trp Ile Ser
Ser Asn Ser Arg Ser Gln Ala Tyr Lys Val Thr Cys Ser 35
40 45Val Arg Gln Ser Ser Ala Gln Lys Arg Lys Tyr
Thr Ile Lys Val Glu 50 55 60Val Pro
Lys Val Ala Thr Gln Thr Val Gly Gly Val Glu Leu Pro Val65
70 75 80Ala Ala Trp Arg Ser Tyr Leu
Asn Met Glu Leu Thr Ile Pro Ile Phe 85 90
95Ala Thr Asn Ser Asp Cys Glu Leu Ile Val Lys Ala Met
Gln Gly Leu 100 105 110Leu Lys
Asp Gly Asn Pro Ile Pro Ser Ala Ile Ala Ala Asn Ser Gly 115
120 125Ile Tyr 13056130PRTEnterobacteria
phage MS2 56Met Ala Ser Asn Phe Thr Gln Phe Val Leu Val Asp Asn Gly Gly
Thr1 5 10 15Gly Asp Val
Thr Val Ala Pro Ser Asn Phe Ala Asn Gly Ile Ala Glu 20
25 30Trp Ile Ser Ser Asn Ser Arg Ser Gln Ala
Tyr Lys Val Thr Cys Ser 35 40
45Val Arg Gln Ser Ser Ala Gln Lys Arg Lys Tyr Thr Ile Lys Val Glu 50
55 60Val Pro Lys Val Ala Thr Gln Thr Val
Gly Gly Val Glu Leu Pro Val65 70 75
80Ala Ala Trp Arg Ser Tyr Leu Asn Met Glu Leu Thr Ile Pro
Ile Phe 85 90 95Ala Thr
Asn Ser Asp Cys Glu Leu Ile Val Lys Ala Met Gln Gly Leu 100
105 110Leu Lys Asp Gly Asn Pro Ile Pro Ser
Ala Ile Ala Ala Asn Ser Gly 115 120
125Ile Tyr 13057121PRTPseudomonas phage PP7 57Lys Thr Ile Val Leu Ser
Val Gly Glu Ala Thr Arg Thr Leu Thr Glu1 5
10 15Ile Gln Ser Thr Ala Asp Arg Gln Ile Phe Glu Glu
Lys Val Gly Pro 20 25 30Leu
Val Gly Arg Leu Arg Leu Thr Ala Ser Leu Arg Gln Asn Gly Ala 35
40 45Lys Thr Ala Tyr Arg Val Asn Leu Lys
Leu Asp Gln Ala Asp Val Val 50 55
60Asp Ser Gly Leu Pro Lys Val Arg Tyr Thr Gln Val Trp Ser His Asp65
70 75 80Val Thr Ile Val Ala
Asn Ser Thr Glu Ala Ser Arg Lys Ser Leu Tyr 85
90 95Asp Leu Thr Lys Ser Leu Val Ala Thr Ser Gln
Val Glu Asp Leu Val 100 105
110Val Asn Leu Val Pro Leu Gly Arg Ser 115
1205862PRTEscherichia virus Mu 58Met Lys Ser Ile Arg Cys Lys Asn Cys Asn
Lys Leu Leu Phe Lys Ala1 5 10
15Asp Ser Phe Asp His Ile Glu Ile Arg Cys Pro Arg Cys Lys Arg His
20 25 30Ile Ile Met Leu Asn Ala
Cys Glu His Pro Thr Glu Lys His Cys Gly 35 40
45Lys Arg Glu Lys Ile Thr His Ser Asp Glu Thr Val Arg Tyr
50 55 6059179PRTArtificial
SequenceDescription of Artificial Sequence Synthetic polypeptide
59Ser Thr Arg Pro Gly Glu Arg Pro Phe Gln Cys Arg Ile Cys Met Arg1
5 10 15Asn Phe Ser Ile Pro Asn
His Leu Ala Arg His Thr Arg Thr His Thr 20 25
30Gly Glu Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Asn
Phe Ser Gln 35 40 45Ser Ala His
Leu Lys Arg His Leu Arg Thr His Thr Gly Glu Lys Pro 50
55 60Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Gln
Asp Val Ser Leu65 70 75
80Val Arg His Leu Lys Thr His Leu Arg Gln Lys Asp Gly Glu Arg Pro
85 90 95Phe Gln Cys Arg Ile Cys
Met Arg Asn Phe Ser Ser Ala Gln Ala Leu 100
105 110Ala Arg His Thr Arg Thr His Thr Gly Glu Lys Pro
Phe Gln Cys Arg 115 120 125Ile Cys
Met Arg Asn Phe Ser Gln Gly Gly Asn Leu Thr Arg His Leu 130
135 140Arg Thr His Thr Gly Glu Lys Pro Phe Gln Cys
Arg Ile Cys Met Arg145 150 155
160Asn Phe Ser Gln His Pro Asn Leu Thr Arg His Leu Lys Thr His Leu
165 170 175Arg Gly
Ser60178PRTArtificial SequenceDescription of Artificial Sequence
Synthetic polypeptide 60Ser Arg Pro Gly Glu Arg Pro Phe Gln Cys Arg
Ile Cys Met Arg Asn1 5 10
15Phe Ser Thr Met Ala Val Leu Arg Arg His Thr Arg Thr His Thr Gly
20 25 30Glu Lys Pro Phe Gln Cys Arg
Ile Cys Met Arg Asn Phe Ser Arg Arg 35 40
45Glu Val Leu Glu Asn His Leu Arg Thr His Thr Gly Glu Lys Pro
Phe 50 55 60Gln Cys Arg Ile Cys Met
Arg Asn Phe Ser Gln Thr Val Asn Leu Asp65 70
75 80Arg His Leu Lys Thr His Leu Arg Gln Lys Asp
Gly Glu Arg Pro Phe 85 90
95Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Lys Lys Asp His Leu His
100 105 110Arg His Thr Arg Thr His
Thr Gly Glu Lys Pro Phe Gln Cys Arg Ile 115 120
125Cys Met Arg Asn Phe Ser Gln Arg Pro His Leu Thr Asn His
Leu Arg 130 135 140Thr His Thr Gly Glu
Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Asn145 150
155 160Phe Ser Val Gly Ala Ser Leu Lys Arg His
Leu Lys Thr His Leu Arg 165 170
175Gly Ser6190PRTArtificial SequenceDescription of Artificial
Sequence Synthetic polypeptide 61Ser Arg Pro Gly Glu Arg Pro Phe Gln
Cys Arg Ile Cys Met Arg Asn1 5 10
15Phe Ser Asp Lys Thr Lys Leu Arg Val His Thr Arg Thr His Thr
Gly 20 25 30Glu Lys Pro Phe
Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Val Arg 35
40 45His Asn Leu Thr Arg His Leu Arg Thr His Thr Gly
Glu Lys Pro Phe 50 55 60Gln Cys Arg
Ile Cys Met Arg Asn Phe Ser Gln Ser Thr Ser Leu Gln65 70
75 80Arg His Leu Lys Thr His Leu Arg
Gly Phe 85 9062176PRTArtificial
SequenceDescription of Artificial Sequence Synthetic polypeptide
62Ser Arg Pro Gly Glu Arg Pro Phe Gln Cys Arg Ile Cys Met Arg Asn1
5 10 15Phe Ser Arg Arg His Gly
Leu Asp Arg His Thr Arg Thr His Thr Gly 20 25
30Glu Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Asn Phe
Ser Asp His 35 40 45Ser Ser Leu
Lys Arg His Leu Arg Thr His Thr Gly Ser Gln Lys Pro 50
55 60Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Val
Arg His Asn Leu65 70 75
80Thr Arg His Leu Arg Thr His Thr Gly Glu Lys Pro Phe Gln Cys Arg
85 90 95Ile Cys Met Arg Asn Phe
Ser Asp His Ser Asn Leu Ser Arg His Leu 100
105 110Lys Thr His Thr Gly Ser Gln Lys Pro Phe Gln Cys
Arg Ile Cys Met 115 120 125Arg Asn
Phe Ser Gln Arg Ser Ser Leu Val Arg His Leu Arg Thr His 130
135 140Thr Gly Glu Lys Pro Phe Gln Cys Arg Ile Cys
Met Arg Asn Phe Ser145 150 155
160Glu Ser Gly His Leu Lys Arg His Leu Arg Thr His Leu Arg Gly Ser
165 170
17563286PRTArtificial SequenceDescription of Artificial Sequence
Synthetic polypeptide 63Ser Arg Pro Gly Glu Arg Pro Phe Gln Cys Arg
Ile Cys Met Arg Asn1 5 10
15Phe Ser Thr Arg Gln Asn Leu Asp Thr His Thr Arg Thr His Thr Gly
20 25 30Glu Lys Pro Phe Gln Cys Arg
Ile Cys Met Arg Asn Phe Ser Arg Arg 35 40
45Asp Thr Leu Glu Arg His Leu Arg Thr His Thr Gly Glu Lys Pro
Phe 50 55 60Gln Cys Arg Ile Cys Met
Arg Asn Phe Ser Arg Pro Asp Ala Leu Pro65 70
75 80Arg His Leu Lys Thr His Leu Arg Gly Ser Gln
Leu Val Lys Ser Glu 85 90
95Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu Lys Tyr Val Pro
100 105 110His Glu Tyr Ile Glu Leu
Ile Glu Ile Ala Arg Asn Ser Thr Gln Asp 115 120
125Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met Lys Val
Tyr Gly 130 135 140Tyr Arg Gly Lys His
Leu Gly Gly Ser Arg Lys Pro Asp Gly Ala Ile145 150
155 160Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly
Val Ile Val Asp Thr Lys 165 170
175Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln Ala Asp Glu Met
180 185 190Gln Arg Tyr Val Glu
Glu Asn Gln Thr Arg Asn Lys His Ile Asn Pro 195
200 205Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr
Glu Phe Lys Phe 210 215 220Leu Phe Val
Ser Gly His Phe Lys Gly Asn Tyr Lys Ala Gln Leu Thr225
230 235 240Arg Leu Asn His Ile Thr Asn
Cys Asn Gly Ala Val Leu Ser Val Glu 245
250 255Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly
Thr Leu Thr Leu 260 265 270Glu
Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile Asn Phe 275
280 28564286PRTArtificial SequenceDescription of
Artificial Sequence Synthetic polypeptide 64Ser Arg Pro Gly Glu Arg
Pro Phe Gln Cys Arg Ile Cys Met Arg Asn1 5
10 15Phe Ser Ser Pro Ser Lys Leu Ile Arg His Thr Arg
Thr His Thr Gly 20 25 30Glu
Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Asp Gly 35
40 45Ser Asn Leu Ala Arg His Leu Arg Thr
His Thr Gly Glu Lys Pro Phe 50 55
60Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Arg Val Asp Asn Leu Pro65
70 75 80Arg His Leu Lys Thr
His Leu Arg Gly Ser Gln Leu Val Lys Ser Glu 85
90 95Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys
Leu Lys Tyr Val Pro 100 105
110His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn Ser Thr Gln Asp
115 120 125Arg Ile Leu Glu Met Lys Val
Met Glu Phe Phe Met Lys Val Tyr Gly 130 135
140Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro Asp Gly Ala
Ile145 150 155 160Tyr Thr
Val Gly Ser Pro Ile Asp Tyr Gly Val Ile Val Asp Thr Lys
165 170 175Ala Tyr Ser Gly Gly Tyr Asn
Leu Pro Ile Gly Gln Ala Asp Glu Met 180 185
190Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys His Ile
Asn Pro 195 200 205Asn Glu Trp Trp
Lys Val Tyr Pro Ser Ser Val Thr Glu Phe Lys Phe 210
215 220Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys
Ala Gln Leu Thr225 230 235
240Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val Leu Ser Val Glu
245 250 255Glu Leu Leu Ile Gly
Gly Glu Met Ile Lys Ala Gly Thr Leu Thr Leu 260
265 270Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile
Asn Phe 275 280
28565196PRTArtificial SequenceDescription of Artificial Sequence
Synthetic polypeptide 65Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys
Ser Glu Leu Arg His1 5 10
15Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala
20 25 30Arg Asn Ser Thr Gln Asp Arg
Ile Leu Glu Met Lys Val Met Glu Phe 35 40
45Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser
Arg 50 55 60Lys Pro Asp Gly Ala Ile
Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly65 70
75 80Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly
Tyr Asn Leu Pro Ile 85 90
95Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg
100 105 110Asn Lys His Ile Asn Pro
Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 115 120
125Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys
Gly Asn 130 135 140Tyr Lys Ala Gln Leu
Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly145 150
155 160Ala Val Leu Ser Val Glu Glu Leu Leu Ile
Gly Gly Glu Met Ile Lys 165 170
175Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly
180 185 190Glu Ile Asn Phe
19566199PRTArtificial SequenceDescription of Artificial Sequence
Synthetic polypeptide 66Val His Asp His Lys Leu Glu Leu Ala Lys Leu
Ile Arg Asn Tyr Glu1 5 10
15Thr Asn Arg Lys Glu Cys Leu Asn Ser Arg Tyr Asn Glu Thr Leu Leu
20 25 30Arg Ser Asp Tyr Leu Asp Pro
Phe Phe Glu Leu Leu Gly Trp Asp Ile 35 40
45Lys Asn Lys Ala Gly Lys Pro Thr Asn Glu Arg Glu Val Val Leu
Glu 50 55 60Glu Ala Leu Lys Ala Ser
Ala Ser Glu His Ser Lys Lys Pro Asp Tyr65 70
75 80Thr Phe Arg Leu Phe Ser Glu Arg Lys Phe Phe
Leu Glu Ala Lys Lys 85 90
95Pro Ser Val His Ile Glu Ser Asp Asn Glu Thr Ala Lys Gln Val Arg
100 105 110Arg Tyr Gly Phe Thr Ala
Lys Leu Lys Ile Ser Val Leu Ser Asn Phe 115 120
125Glu Tyr Leu Val Ile Tyr Asp Thr Ser Val Lys Val Asp Gly
Asp Asp 130 135 140Thr Phe Asn Lys Ala
Arg Ile Lys Lys Tyr His Tyr Thr Glu Tyr Glu145 150
155 160Thr His Phe Asp Glu Ile Cys Asp Leu Leu
Gly Arg Glu Ser Val Tyr 165 170
175Ser Gly Asn Phe Asp Lys Glu Trp Leu Ser Ile Glu Asn Lys Ile Asn
180 185 190His Phe Ser Val Asp
Thr Leu 19567119PRTArtificial SequenceDescription of Artificial
Sequence Synthetic polypeptide 67Tyr Asn Glu Thr Leu Leu Arg Ser Asp
Tyr Leu Asp Pro Phe Phe Glu1 5 10
15Leu Leu Gly Trp Asp Ile Lys Asn Lys Ala Gly Lys Pro Thr Asn
Glu 20 25 30Arg Glu Val Val
Leu Glu Glu Ala Leu Lys Ala Ser Ala Ser Glu His 35
40 45Ser Lys Lys Pro Asp Tyr Thr Phe Arg Leu Phe Ser
Glu Arg Lys Phe 50 55 60Phe Leu Glu
Ala Lys Lys Pro Ser Val His Ile Glu Ser Asp Asn Glu65 70
75 80Thr Ala Lys Gln Val Arg Arg Tyr
Gly Phe Thr Ala Lys Leu Lys Ile 85 90
95Ser Val Leu Ser Asn Phe Glu Tyr Leu Val Ile Tyr Asp Thr
Ser Val 100 105 110Lys Val Asp
Gly Asp Asp Thr 11568972PRTRuminococcus flavefaciens 68Glu Ala Ser
Ile Glu Lys Lys Lys Ser Phe Ala Lys Gly Met Gly Val1 5
10 15Lys Ser Thr Leu Val Ser Gly Ser Lys
Val Tyr Met Thr Thr Phe Ala 20 25
30Glu Gly Ser Asp Ala Arg Leu Glu Lys Ile Val Glu Gly Asp Ser Ile
35 40 45Arg Ser Val Asn Glu Gly Glu
Ala Phe Ser Ala Glu Met Ala Asp Lys 50 55
60Asn Ala Gly Tyr Lys Ile Gly Asn Ala Lys Phe Ser His Pro Lys Gly65
70 75 80Tyr Ala Val Val
Ala Asn Asn Pro Leu Tyr Thr Gly Pro Val Gln Gln 85
90 95Asp Met Leu Gly Leu Lys Glu Thr Leu Glu
Lys Arg Tyr Phe Gly Glu 100 105
110Ser Ala Asp Gly Asn Asp Asn Ile Cys Ile Gln Val Ile His Asn Ile
115 120 125Leu Asp Ile Glu Lys Ile Leu
Ala Glu Tyr Ile Thr Asn Ala Ala Tyr 130 135
140Ala Val Asn Asn Ile Ser Gly Leu Asp Lys Asp Ile Ile Gly Phe
Gly145 150 155 160Lys Phe
Ser Thr Val Tyr Thr Tyr Asp Glu Phe Lys Asp Pro Glu His
165 170 175His Arg Ala Ala Phe Asn Asn
Asn Asp Lys Leu Ile Asn Ala Ile Lys 180 185
190Ala Gln Tyr Asp Glu Phe Asp Asn Phe Leu Asp Asn Pro Arg
Leu Gly 195 200 205Tyr Phe Gly Gln
Ala Phe Phe Ser Lys Glu Gly Arg Asn Tyr Ile Ile 210
215 220Asn Tyr Gly Asn Glu Cys Tyr Asp Ile Leu Ala Leu
Leu Ser Gly Leu225 230 235
240Arg His Trp Val Val His Asn Asn Glu Glu Glu Ser Arg Ile Ser Arg
245 250 255Thr Trp Leu Tyr Asn
Leu Asp Lys Asn Leu Asp Asn Glu Tyr Ile Ser 260
265 270Thr Leu Asn Tyr Leu Tyr Asp Arg Ile Thr Asn Glu
Leu Thr Asn Ser 275 280 285Phe Ser
Lys Asn Ser Ala Ala Asn Val Asn Tyr Ile Ala Glu Thr Leu 290
295 300Gly Ile Asn Pro Ala Glu Phe Ala Glu Gln Tyr
Phe Arg Phe Ser Ile305 310 315
320Met Lys Glu Gln Lys Asn Leu Gly Phe Asn Ile Thr Lys Leu Arg Glu
325 330 335Val Met Leu Asp
Arg Lys Asp Met Ser Glu Ile Arg Lys Asn His Lys 340
345 350Val Phe Asp Ser Ile Arg Thr Lys Val Tyr Thr
Met Met Asp Phe Val 355 360 365Ile
Tyr Arg Tyr Tyr Ile Glu Glu Asp Ala Lys Val Ala Ala Ala Asn 370
375 380Lys Ser Leu Pro Asp Asn Glu Lys Ser Leu
Ser Glu Lys Asp Ile Phe385 390 395
400Val Ile Asn Leu Arg Gly Ser Phe Asn Asp Asp Gln Lys Asp Ala
Leu 405 410 415Tyr Tyr Asp
Glu Ala Asn Arg Ile Trp Arg Lys Leu Glu Asn Ile Met 420
425 430His Asn Ile Lys Glu Phe Arg Gly Asn Lys
Thr Arg Glu Tyr Lys Lys 435 440
445Lys Asp Ala Pro Arg Leu Pro Arg Ile Leu Pro Ala Gly Arg Asp Val 450
455 460Ser Ala Phe Ser Lys Leu Met Tyr
Ala Leu Thr Met Phe Leu Asp Gly465 470
475 480Lys Glu Ile Asn Asp Leu Leu Thr Thr Leu Ile Asn
Lys Phe Asp Asn 485 490
495Ile Gln Ser Phe Leu Lys Val Met Pro Leu Ile Gly Val Asn Ala Lys
500 505 510Phe Val Glu Glu Tyr Ala
Phe Phe Lys Asp Ser Ala Lys Ile Ala Asp 515 520
525Glu Leu Arg Leu Ile Lys Ser Phe Ala Arg Met Gly Glu Pro
Ile Ala 530 535 540Asp Ala Arg Arg Ala
Met Tyr Ile Asp Ala Ile Arg Ile Leu Gly Thr545 550
555 560Asn Leu Ser Tyr Asp Glu Leu Lys Ala Leu
Ala Asp Thr Phe Ser Leu 565 570
575Asp Glu Asn Gly Asn Lys Leu Lys Lys Gly Lys His Gly Met Arg Asn
580 585 590Phe Ile Ile Asn Asn
Val Ile Ser Asn Lys Arg Phe His Tyr Leu Ile 595
600 605Arg Tyr Gly Asp Pro Ala His Leu His Glu Ile Ala
Lys Asn Glu Ala 610 615 620Val Val Lys
Phe Val Leu Gly Arg Ile Ala Asp Ile Gln Lys Lys Gln625
630 635 640Gly Gln Asn Gly Lys Asn Gln
Ile Asp Arg Tyr Tyr Glu Thr Cys Ile 645
650 655Gly Lys Asp Lys Gly Lys Ser Val Ser Glu Lys Val
Asp Ala Leu Thr 660 665 670Lys
Ile Ile Thr Gly Met Asn Tyr Asp Gln Phe Asp Lys Lys Arg Ser 675
680 685Val Ile Glu Asp Thr Gly Arg Glu Asn
Ala Glu Arg Glu Lys Phe Lys 690 695
700Lys Ile Ile Ser Leu Tyr Leu Thr Val Ile Tyr His Ile Leu Lys Asn705
710 715 720Ile Val Asn Ile
Asn Ala Arg Tyr Val Ile Gly Phe His Cys Val Glu 725
730 735Arg Asp Ala Gln Leu Tyr Lys Glu Lys Gly
Tyr Asp Ile Asn Leu Lys 740 745
750Lys Leu Glu Glu Lys Gly Phe Ser Ser Val Thr Lys Leu Cys Ala Gly
755 760 765Ile Asp Glu Thr Ala Pro Asp
Lys Arg Lys Asp Val Glu Lys Glu Met 770 775
780Ala Glu Arg Ala Lys Glu Ser Ile Asp Ser Leu Glu Ser Ala Asn
Pro785 790 795 800Lys Leu
Tyr Ala Asn Tyr Ile Lys Tyr Ser Asp Glu Lys Lys Ala Glu
805 810 815Glu Phe Thr Arg Gln Ile Asn
Arg Glu Lys Ala Lys Thr Ala Leu Asn 820 825
830Ala Tyr Leu Arg Asn Thr Lys Trp Asn Val Ile Ile Arg Glu
Asp Leu 835 840 845Leu Arg Ile Asp
Asn Lys Thr Cys Thr Leu Phe Arg Asn Lys Ala Val 850
855 860His Leu Glu Val Ala Arg Tyr Val His Ala Tyr Ile
Asn Asp Ile Ala865 870 875
880Glu Val Asn Ser Tyr Phe Gln Leu Tyr His Tyr Ile Met Gln Arg Ile
885 890 895Ile Met Asn Glu Arg
Tyr Glu Lys Ser Ser Gly Lys Val Ser Glu Tyr 900
905 910Phe Asp Ala Val Asn Asp Glu Lys Lys Tyr Asn Asp
Arg Leu Leu Lys 915 920 925Leu Leu
Cys Val Pro Phe Gly Tyr Cys Ile Pro Arg Phe Lys Asn Leu 930
935 940Ser Ile Glu Ala Leu Phe Asp Arg Asn Glu Ala
Ala Lys Phe Asp Lys945 950 955
960Glu Lys Lys Lys Val Ser Gly Asn Ser Gly Ser Gly
965 97069991PRTArtificial SequenceDescription of
Artificial Sequence Synthetic polypeptide 69Glu Ala Ser Ile Glu Lys
Lys Lys Ser Phe Ala Lys Gly Met Gly Val1 5
10 15Lys Ser Thr Leu Val Ser Gly Ser Lys Val Tyr Met
Thr Thr Phe Ala 20 25 30Glu
Gly Ser Asp Ala Arg Leu Glu Lys Ile Val Glu Gly Asp Ser Ile 35
40 45Arg Ser Val Asn Glu Gly Glu Ala Phe
Ser Ala Glu Met Ala Asp Lys 50 55
60Asn Ala Gly Tyr Lys Ile Gly Asn Ala Lys Phe Ser His Pro Lys Gly65
70 75 80Tyr Ala Val Val Ala
Asn Asn Pro Leu Tyr Thr Gly Pro Val Gln Gln 85
90 95Asp Met Leu Gly Leu Lys Glu Thr Leu Glu Lys
Arg Tyr Phe Gly Glu 100 105
110Ser Ala Asp Gly Asn Asp Asn Ile Cys Ile Gln Val Ile His Asn Ile
115 120 125Leu Asp Ile Glu Lys Ile Leu
Ala Glu Tyr Ile Thr Asn Ala Ala Tyr 130 135
140Ala Val Asn Asn Ile Ser Gly Leu Asp Lys Asp Ile Ile Gly Phe
Gly145 150 155 160Lys Phe
Ser Thr Val Tyr Thr Tyr Asp Glu Phe Lys Asp Pro Glu His
165 170 175His Arg Ala Ala Phe Asn Asn
Asn Asp Lys Leu Ile Asn Ala Ile Lys 180 185
190Ala Gln Tyr Asp Glu Phe Asp Asn Phe Leu Asp Asn Pro Arg
Leu Gly 195 200 205Tyr Phe Gly Gln
Ala Phe Phe Ser Lys Glu Gly Arg Asn Tyr Ile Ile 210
215 220Asn Tyr Gly Asn Glu Cys Tyr Asp Ile Leu Ala Leu
Leu Ser Gly Leu225 230 235
240Ala His Trp Val Val Ala Asn Asn Glu Glu Glu Ser Arg Ile Ser Arg
245 250 255Thr Trp Leu Tyr Asn
Leu Asp Lys Asn Leu Asp Asn Glu Tyr Ile Ser 260
265 270Thr Leu Asn Tyr Leu Tyr Asp Arg Ile Thr Asn Glu
Leu Thr Asn Ser 275 280 285Phe Ser
Lys Asn Ser Ala Ala Asn Val Asn Tyr Ile Ala Glu Thr Leu 290
295 300Gly Ile Asn Pro Ala Glu Phe Ala Glu Gln Tyr
Phe Arg Phe Ser Ile305 310 315
320Met Lys Glu Gln Lys Asn Leu Gly Phe Asn Ile Thr Lys Leu Arg Glu
325 330 335Val Met Leu Asp
Arg Lys Asp Met Ser Glu Ile Arg Lys Asn His Lys 340
345 350Val Phe Asp Ser Ile Arg Thr Lys Val Tyr Thr
Met Met Asp Phe Val 355 360 365Ile
Tyr Arg Tyr Tyr Ile Glu Glu Asp Ala Lys Val Ala Ala Ala Asn 370
375 380Lys Ser Leu Pro Asp Asn Glu Lys Ser Leu
Ser Glu Lys Asp Ile Phe385 390 395
400Val Ile Asn Leu Arg Gly Ser Phe Asn Asp Asp Gln Lys Asp Ala
Leu 405 410 415Tyr Tyr Asp
Glu Ala Asn Arg Ile Trp Arg Lys Leu Glu Asn Ile Met 420
425 430His Asn Ile Lys Glu Phe Arg Gly Asn Lys
Thr Arg Glu Tyr Lys Lys 435 440
445Lys Asp Ala Pro Arg Leu Pro Arg Ile Leu Pro Ala Gly Arg Asp Val 450
455 460Ser Ala Phe Ser Lys Leu Met Tyr
Ala Leu Thr Met Phe Leu Asp Gly465 470
475 480Lys Glu Ile Asn Asp Leu Leu Thr Thr Leu Ile Asn
Lys Phe Asp Asn 485 490
495Ile Gln Ser Phe Leu Lys Val Met Pro Leu Ile Gly Val Asn Ala Lys
500 505 510Phe Val Glu Glu Tyr Ala
Phe Phe Lys Asp Ser Ala Lys Ile Ala Asp 515 520
525Glu Leu Arg Leu Ile Lys Ser Phe Ala Arg Met Gly Glu Pro
Ile Ala 530 535 540Asp Ala Arg Arg Ala
Met Tyr Ile Asp Ala Ile Arg Ile Leu Gly Thr545 550
555 560Asn Leu Ser Tyr Asp Glu Leu Lys Ala Leu
Ala Asp Thr Phe Ser Leu 565 570
575Asp Glu Asn Gly Asn Lys Leu Lys Lys Gly Lys His Gly Met Arg Asn
580 585 590Phe Ile Ile Asn Asn
Val Ile Ser Asn Lys Arg Phe His Tyr Leu Ile 595
600 605Arg Tyr Gly Asp Pro Ala His Leu His Glu Ile Ala
Lys Asn Glu Ala 610 615 620Val Val Lys
Phe Val Leu Gly Arg Ile Ala Asp Ile Gln Lys Lys Gln625
630 635 640Gly Gln Asn Gly Lys Asn Gln
Ile Asp Arg Tyr Tyr Glu Thr Cys Ile 645
650 655Gly Lys Asp Lys Gly Lys Ser Val Ser Glu Lys Val
Asp Ala Leu Thr 660 665 670Lys
Ile Ile Thr Gly Met Asn Tyr Asp Gln Phe Asp Lys Lys Arg Ser 675
680 685Val Ile Glu Asp Thr Gly Arg Glu Asn
Ala Glu Arg Glu Lys Phe Lys 690 695
700Lys Ile Ile Ser Leu Tyr Leu Thr Val Ile Tyr His Ile Leu Lys Asn705
710 715 720Ile Val Asn Ile
Asn Ala Arg Tyr Val Ile Gly Phe His Cys Val Glu 725
730 735Arg Asp Ala Gln Leu Tyr Lys Glu Lys Gly
Tyr Asp Ile Asn Leu Lys 740 745
750Lys Leu Glu Glu Lys Gly Phe Ser Ser Val Thr Lys Leu Cys Ala Gly
755 760 765Ile Asp Glu Thr Ala Pro Asp
Lys Arg Lys Asp Val Glu Lys Glu Met 770 775
780Ala Glu Arg Ala Lys Glu Ser Ile Asp Ser Leu Glu Ser Ala Asn
Pro785 790 795 800Lys Leu
Tyr Ala Asn Tyr Ile Lys Tyr Ser Asp Glu Lys Lys Ala Glu
805 810 815Glu Phe Thr Arg Gln Ile Asn
Arg Glu Lys Ala Lys Thr Ala Leu Asn 820 825
830Ala Tyr Leu Arg Asn Thr Lys Trp Asn Val Ile Ile Arg Glu
Asp Leu 835 840 845Leu Arg Ile Asp
Asn Lys Thr Cys Thr Leu Phe Ala Asn Lys Ala Val 850
855 860Ala Leu Glu Val Ala Arg Tyr Val His Ala Tyr Ile
Asn Asp Ile Ala865 870 875
880Glu Val Asn Ser Tyr Phe Gln Leu Tyr His Tyr Ile Met Gln Arg Ile
885 890 895Ile Met Asn Glu Arg
Tyr Glu Lys Ser Ser Gly Lys Val Ser Glu Tyr 900
905 910Phe Asp Ala Val Asn Asp Glu Lys Lys Tyr Asn Asp
Arg Leu Leu Lys 915 920 925Leu Leu
Cys Val Pro Phe Gly Tyr Cys Ile Pro Arg Phe Lys Asn Leu 930
935 940Ser Ile Glu Ala Leu Phe Asp Arg Asn Glu Ala
Ala Lys Phe Asp Lys945 950 955
960Glu Lys Lys Lys Val Ser Gly Asn Ser Gly Ser Gly Pro Lys Lys Lys
965 970 975Arg Lys Val Ala
Ala Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala 980
985 9907025PRTArtificial SequenceDescription of
Artificial Sequence Synthetic peptide 70Ile Trp Leu Thr Ala Leu Lys
Phe Leu Gly Lys His Ala Ala Lys His1 5 10
15Glu Ala Lys Gln Gln Leu Ser Lys Leu 20
257175PRTArtificial SequenceDescription of Artificial
Sequence Synthetic polypeptide 71Ile Trp Leu Thr Ala Leu Lys Phe Leu
Gly Lys His Ala Ala Lys His1 5 10
15Glu Ala Lys Gln Gln Leu Ser Lys Leu Asn Ala Val Gly Gln Asp
Thr 20 25 30Gln Glu Val Ile
Val Val Pro His Ser Leu Pro Phe Lys Val Val Val 35
40 45Ile Ser Ala Ile Leu Ala Leu Val Val Leu Thr Ile
Ile Ser Leu Ile 50 55 60Ile Leu Ile
Met Leu Trp Gln Lys Lys Pro Arg65 70
757230PRTArtificial SequenceDescription of Artificial Sequence Synthetic
polypeptide 72Trp Glu Ala Lys Leu Ala Lys Ala Leu Ala Lys Ala Leu Ala
Lys His1 5 10 15Leu Ala
Lys Ala Leu Ala Lys Ala Leu Lys Ala Cys Glu Ala 20
25 307380PRTArtificial SequenceDescription of
Artificial Sequence Synthetic polypeptide 73Trp Glu Ala Lys Leu Ala
Lys Ala Leu Ala Lys Ala Leu Ala Lys His1 5
10 15Leu Ala Lys Ala Leu Ala Lys Ala Leu Lys Ala Cys
Glu Ala Asn Ala 20 25 30Val
Gly Gln Asp Thr Gln Glu Val Ile Val Val Pro His Ser Leu Pro 35
40 45Phe Lys Val Val Val Ile Ser Ala Ile
Leu Ala Leu Val Val Leu Thr 50 55
60Ile Ile Ser Leu Ile Ile Leu Ile Met Leu Trp Gln Lys Lys Pro Arg65
70 75 807426PRTArtificial
SequenceDescription of Artificial Sequence Synthetic peptide 74Lys
Lys Ala Leu Leu His Ala Ala Leu Ala His Leu Leu Ala Leu Ala1
5 10 15His His Leu Leu Ala Leu Leu
Lys Lys Ala 20 257576PRTArtificial
SequenceDescription of Artificial Sequence Synthetic polypeptide
75Lys Lys Ala Leu Leu His Ala Ala Leu Ala His Leu Leu Ala Leu Ala1
5 10 15His His Leu Leu Ala Leu
Leu Lys Lys Ala Asn Ala Val Gly Gln Asp 20 25
30Thr Gln Glu Val Ile Val Val Pro His Ser Leu Pro Phe
Lys Val Val 35 40 45Val Ile Ser
Ala Ile Leu Ala Leu Val Val Leu Thr Ile Ile Ser Leu 50
55 60Ile Ile Leu Ile Met Leu Trp Gln Lys Lys Pro Arg65
70 757650PRTArtificial
SequenceDescription of Artificial Sequence Synthetic polypeptide
76Asn Ala Val Gly Gln Asp Thr Gln Glu Val Ile Val Val Pro His Ser1
5 10 15Leu Pro Phe Lys Val Val
Val Ile Ser Ala Ile Leu Ala Leu Val Val 20 25
30Leu Thr Ile Ile Ser Leu Ile Ile Leu Ile Met Leu Trp
Gln Lys Lys 35 40 45Pro Arg
507734PRTLactococcus lactis 77Ile Thr Ser Ile Ser Leu Cys Thr Pro Gly Cys
Lys Thr Gly Ala Leu1 5 10
15Met Gly Cys Asn Met Lys Thr Ala Thr Cys His Cys Ser Ile His Val
20 25 30Ser Lys7834PRTLactococcus
lactis 78Ile Thr Ser Ile Ser Leu Cys Thr Pro Gly Cys Lys Thr Gly Ala Leu1
5 10 15Met Gly Cys Asn
Met Lys Thr Ala Thr Cys Asn Cys Ser Ile His Val 20
25 30Ser Lys7934PRTLactococcus lactis 79Ile Thr Ser
Ile Ser Leu Cys Thr Pro Gly Cys Lys Thr Gly Ala Leu1 5
10 15Met Gly Cys Asn Met Lys Thr Ala Thr
Cys Asn Cys Ser Val His Val 20 25
30Ser Lys8034PRTLactococcus lactis 80Ile Thr Ser Ile Ser Leu Cys Thr
Pro Gly Cys Lys Thr Gly Val Leu1 5 10
15Met Gly Cys Asn Leu Lys Thr Ala Thr Cys Asn Cys Ser Val
His Val 20 25 30Ser
Lys8134PRTStreptococcus hyointestinalis 81Phe Thr Ser Ile Ser Met Cys Thr
Pro Gly Cys Lys Thr Gly Ala Leu1 5 10
15Met Thr Cys Asn Tyr Lys Thr Ala Thr Cys His Cys Ser Ile
Lys Val 20 25 30Ser
Lys8231PRTStreptococcus uberis 82Ile Thr Ser Lys Ser Leu Cys Thr Pro Gly
Cys Lys Thr Gly Ile Leu1 5 10
15Met Thr Cys Pro Leu Lys Thr Ala Thr Cys Gly Cys His Phe Gly
20 25 308331PRTStreptococcus uberis
83Val Thr Ser Lys Ser Leu Cys Thr Pro Gly Cys Lys Thr Gly Ile Leu1
5 10 15Met Thr Cys Pro Leu Lys
Thr Ala Thr Cys Gly Cys His Phe Gly 20 25
308431PRTStreptococcus galloyticus 84Val Thr Ser Lys Ser Leu
Cys Thr Pro Gly Cys Lys Thr Gly Ile Leu1 5
10 15Met Thr Cys Ala Ile Lys Thr Ala Thr Cys Gly Cys
His Phe Gly 20 25
308534PRTLactococcus lactis 85Ile Thr Ser Ile Ser Leu Cys Thr Pro Gly Cys
Lys Thr Gly Ala Leu1 5 10
15Met Gly Cys Asn Met Lys Thr Ala Thr Cys His Cys Ala Ile His Val
20 25 30Ser Lys8634PRTLactococcus
lactis 86Ile Thr Ser Ile Ser Leu Cys Thr Pro Gly Cys Lys Thr Gly Ala Leu1
5 10 15Met Gly Cys Asn
Met Lys Thr Ala Thr Cys His Cys Asp Ile His Val 20
25 30Ser Lys8734PRTLactococcus lactis 87Ile Thr Ser
Ile Ser Leu Cys Thr Pro Gly Cys Lys Thr Gly Ala Leu1 5
10 15Met Gly Cys Asn Met Lys Thr Ala Thr
Cys His Cys Glu Ile His Val 20 25
30Ser Lys8834PRTLactococcus lactis 88Ile Thr Ser Ile Ser Leu Cys Thr
Pro Gly Cys Lys Thr Gly Ala Leu1 5 10
15Met Gly Cys Asn Met Lys Thr Ala Thr Cys His Cys Gly Ile
His Val 20 25 30Ser
Lys8934PRTLactococcus lactis 89Ile Thr Ser Ile Ser Leu Cys Thr Pro Gly
Cys Lys Thr Gly Ala Leu1 5 10
15Met Gly Cys Asn Met Thr Thr Ala Thr Cys His Cys Ser Ile His Val
20 25 30Ser
Lys9034PRTLactococcus lactis 90Ile Thr Ser Ile Ser Leu Cys Thr Pro Gly
Cys Lys Thr Gly Ala Leu1 5 10
15Met Gly Cys Pro Met Lys Thr Ala Thr Cys His Cys Ser Ile His Val
20 25 30Ser
Lys9134PRTLactococcus lactis 91Ile Thr Ser Ile Ser Leu Cys Thr Pro Gly
Cys Lys Thr Gly Ala Leu1 5 10
15Met Gly Cys Asn Val Lys Thr Ala Thr Cys His Cys Ser Ile His Val
20 25 30Ser
Lys9234PRTLactococcus lactis 92Ile Thr Ser Ile Ser Leu Cys Thr Pro Gly
Cys Lys Thr Gly Ala Leu1 5 10
15Met Gly Cys Asn Met Ser Thr Ala Thr Cys His Cys Ser Ile His Val
20 25 30Ser
Lys9334PRTLactococcus lactis 93Ile Thr Ser Ile Ser Leu Cys Thr Pro Gly
Cys Lys Thr Gly Ala Leu1 5 10
15Met Gly Cys Lys Met Lys Thr Ala Thr Cys Asn Cys Ser Ile His Val
20 25 30Ser
Lys9434PRTLactococcus lactis 94Ile Thr Ser Ile Ser Leu Cys Thr Pro Gly
Cys Lys Thr Gly Ala Leu1 5 10
15Met Gly Cys Asn Lys Lys Thr Ala Thr Cys Asn Cys Ser Ile His Val
20 25 30Ser
Lys95397PRTAdeno-associated virus - 2 95Met Glu Leu Val Gly Trp Leu Val
Asp Lys Gly Ile Thr Ser Glu Lys1 5 10
15Gln Trp Ile Gln Glu Asp Gln Ala Ser Tyr Ile Ser Phe Asn
Ala Ala 20 25 30Ser Asn Ser
Arg Ser Gln Ile Lys Ala Ala Leu Asp Asn Ala Gly Lys 35
40 45Ile Met Ser Leu Thr Lys Thr Ala Pro Asp Tyr
Leu Val Gly Gln Gln 50 55 60Pro Val
Glu Asp Ile Ser Ser Asn Arg Ile Tyr Lys Ile Leu Glu Leu65
70 75 80Asn Gly Tyr Asp Pro Gln Tyr
Ala Ala Ser Val Phe Leu Gly Trp Ala 85 90
95Thr Lys Lys Phe Gly Lys Arg Asn Thr Ile Trp Leu Phe
Gly Pro Ala 100 105 110Thr Thr
Gly Lys Thr Asn Ile Ala Glu Ala Ile Ala His Thr Val Pro 115
120 125Phe Tyr Gly Cys Val Asn Trp Thr Asn Glu
Asn Phe Pro Phe Asn Asp 130 135 140Cys
Val Asp Lys Met Val Ile Trp Trp Glu Glu Gly Lys Met Thr Ala145
150 155 160Lys Val Val Glu Ser Ala
Lys Ala Ile Leu Gly Gly Ser Lys Val Arg 165
170 175Val Asp Gln Lys Cys Lys Ser Ser Ala Gln Ile Asp
Pro Thr Pro Val 180 185 190Ile
Val Thr Ser Asn Thr Asn Met Cys Ala Val Ile Asp Gly Asn Ser 195
200 205Thr Thr Phe Glu His Gln Gln Pro Leu
Gln Asp Arg Met Phe Lys Phe 210 215
220Glu Leu Thr Arg Arg Leu Asp His Asp Phe Gly Lys Val Thr Lys Gln225
230 235 240Glu Val Lys Asp
Phe Phe Arg Trp Ala Lys Asp His Val Val Glu Val 245
250 255Glu His Glu Phe Tyr Val Lys Lys Gly Gly
Ala Lys Lys Arg Pro Ala 260 265
270Pro Ser Asp Ala Asp Ile Ser Glu Pro Lys Arg Val Arg Glu Ser Val
275 280 285Ala Gln Pro Ser Thr Ser Asp
Ala Glu Ala Ser Ile Asn Tyr Ala Asp 290 295
300Arg Tyr Gln Asn Lys Cys Ser Arg His Val Gly Met Asn Leu Met
Leu305 310 315 320Phe Pro
Cys Arg Gln Cys Glu Arg Met Asn Gln Asn Ser Asn Ile Cys
325 330 335Phe Thr His Gly Gln Lys Asp
Cys Leu Glu Cys Phe Pro Val Ser Glu 340 345
350Ser Gln Pro Val Ser Val Val Lys Lys Ala Tyr Gln Lys Leu
Cys Tyr 355 360 365Ile His His Ile
Met Gly Lys Val Pro Asp Ala Cys Thr Ala Cys Asp 370
375 380Leu Val Asn Val Asp Leu Asp Asp Cys Ile Phe Glu
Gln385 390 39596621PRTAdeno-associated
virus - 2 96Met Pro Gly Phe Tyr Glu Ile Val Ile Lys Val Pro Ser Asp Leu
Asp1 5 10 15Glu His Leu
Pro Gly Ile Ser Asp Ser Phe Val Asn Trp Val Ala Glu 20
25 30Lys Glu Trp Glu Leu Pro Pro Asp Ser Asp
Met Asp Leu Asn Leu Ile 35 40
45Glu Gln Ala Pro Leu Thr Val Ala Glu Lys Leu Gln Arg Asp Phe Leu 50
55 60Thr Glu Trp Arg Arg Val Ser Lys Ala
Pro Glu Ala Leu Phe Phe Val65 70 75
80Gln Phe Glu Lys Gly Glu Ser Tyr Phe His Met His Val Leu
Val Glu 85 90 95Thr Thr
Gly Val Lys Ser Met Val Leu Gly Arg Phe Leu Ser Gln Ile 100
105 110Arg Glu Lys Leu Ile Gln Arg Ile Tyr
Arg Gly Ile Glu Pro Thr Leu 115 120
125Pro Asn Trp Phe Ala Val Thr Lys Thr Arg Asn Gly Ala Gly Gly Gly
130 135 140Asn Lys Val Val Asp Glu Cys
Tyr Ile Pro Asn Tyr Leu Leu Pro Lys145 150
155 160Thr Gln Pro Glu Leu Gln Trp Ala Trp Thr Asn Met
Glu Gln Tyr Leu 165 170
175Ser Ala Cys Leu Asn Leu Thr Glu Arg Lys Arg Leu Val Ala Gln His
180 185 190Leu Thr His Val Ser Gln
Thr Gln Glu Gln Asn Lys Glu Asn Gln Asn 195 200
205Pro Asn Ser Asp Ala Pro Val Ile Arg Ser Lys Thr Ser Ala
Arg Tyr 210 215 220Met Glu Leu Val Gly
Trp Leu Val Asp Lys Gly Ile Thr Ser Glu Lys225 230
235 240Gln Trp Ile Gln Glu Asp Gln Ala Ser Tyr
Ile Ser Phe Asn Ala Ala 245 250
255Ser Asn Ser Arg Ser Gln Ile Lys Ala Ala Leu Asp Asn Ala Gly Lys
260 265 270Ile Met Ser Leu Thr
Lys Thr Ala Pro Asp Tyr Leu Val Gly Gln Gln 275
280 285Pro Val Glu Asp Ile Ser Ser Asn Arg Ile Tyr Lys
Ile Leu Glu Leu 290 295 300Asn Gly Tyr
Asp Pro Gln Tyr Ala Ala Ser Val Phe Leu Gly Trp Ala305
310 315 320Thr Lys Lys Phe Gly Lys Arg
Asn Thr Ile Trp Leu Phe Gly Pro Ala 325
330 335Thr Thr Gly Lys Thr Asn Ile Ala Glu Ala Ile Ala
His Thr Val Pro 340 345 350Phe
Tyr Gly Cys Val Asn Trp Thr Asn Glu Asn Phe Pro Phe Asn Asp 355
360 365Cys Val Asp Lys Met Val Ile Trp Trp
Glu Glu Gly Lys Met Thr Ala 370 375
380Lys Val Val Glu Ser Ala Lys Ala Ile Leu Gly Gly Ser Lys Val Arg385
390 395 400Val Asp Gln Lys
Cys Lys Ser Ser Ala Gln Ile Asp Pro Thr Pro Val 405
410 415Ile Val Thr Ser Asn Thr Asn Met Cys Ala
Val Ile Asp Gly Asn Ser 420 425
430Thr Thr Phe Glu His Gln Gln Pro Leu Gln Asp Arg Met Phe Lys Phe
435 440 445Glu Leu Thr Arg Arg Leu Asp
His Asp Phe Gly Lys Val Thr Lys Gln 450 455
460Glu Val Lys Asp Phe Phe Arg Trp Ala Lys Asp His Val Val Glu
Val465 470 475 480Glu His
Glu Phe Tyr Val Lys Lys Gly Gly Ala Lys Lys Arg Pro Ala
485 490 495Pro Ser Asp Ala Asp Ile Ser
Glu Pro Lys Arg Val Arg Glu Ser Val 500 505
510Ala Gln Pro Ser Thr Ser Asp Ala Glu Ala Ser Ile Asn Tyr
Ala Asp 515 520 525Arg Tyr Gln Asn
Lys Cys Ser Arg His Val Gly Met Asn Leu Met Leu 530
535 540Phe Pro Cys Arg Gln Cys Glu Arg Met Asn Gln Asn
Ser Asn Ile Cys545 550 555
560Phe Thr His Gly Gln Lys Asp Cys Leu Glu Cys Phe Pro Val Ser Glu
565 570 575Ser Gln Pro Val Ser
Val Val Lys Lys Ala Tyr Gln Lys Leu Cys Tyr 580
585 590Ile His His Ile Met Gly Lys Val Pro Asp Ala Cys
Thr Ala Cys Asp 595 600 605Leu Val
Asn Val Asp Leu Asp Asp Cys Ile Phe Glu Gln 610 615
62097735PRTAdeno-associated virus - 2 97Met Ala Ala Asp Gly
Tyr Leu Pro Asp Trp Leu Glu Asp Thr Leu Ser1 5
10 15Glu Gly Ile Arg Gln Trp Trp Lys Leu Lys Pro
Gly Pro Pro Pro Pro 20 25
30Lys Pro Ala Glu Arg His Lys Asp Asp Ser Arg Gly Leu Val Leu Pro
35 40 45Gly Tyr Lys Tyr Leu Gly Pro Phe
Asn Gly Leu Asp Lys Gly Glu Pro 50 55
60Val Asn Glu Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp65
70 75 80Arg Gln Leu Asp Ser
Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala 85
90 95Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp
Thr Ser Phe Gly Gly 100 105
110Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro
115 120 125Leu Gly Leu Val Glu Glu Pro
Val Lys Thr Ala Pro Gly Lys Lys Arg 130 135
140Pro Val Glu His Ser Pro Val Glu Pro Asp Ser Ser Ser Gly Thr
Gly145 150 155 160Lys Ala
Gly Gln Gln Pro Ala Arg Lys Arg Leu Asn Phe Gly Gln Thr
165 170 175Gly Asp Ala Asp Ser Val Pro
Asp Pro Gln Pro Leu Gly Gln Pro Pro 180 185
190Ala Ala Pro Ser Gly Leu Gly Thr Asn Thr Met Ala Thr Gly
Ser Gly 195 200 205Ala Pro Met Ala
Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ser 210
215 220Ser Gly Asn Trp His Cys Asp Ser Thr Trp Met Gly
Asp Arg Val Ile225 230 235
240Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu
245 250 255Tyr Lys Gln Ile Ser
Ser Gln Ser Gly Ala Ser Asn Asp Asn His Tyr 260
265 270Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe
Asn Arg Phe His 275 280 285Cys His
Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn Trp 290
295 300Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu
Phe Asn Ile Gln Val305 310 315
320Lys Glu Val Thr Gln Asn Asp Gly Thr Thr Thr Ile Ala Asn Asn Leu
325 330 335Thr Ser Thr Val
Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu Pro Tyr 340
345 350Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro
Pro Phe Pro Ala Asp 355 360 365Val
Phe Met Val Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly Ser 370
375 380Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys
Leu Glu Tyr Phe Pro Ser385 390 395
400Gln Met Leu Arg Thr Gly Asn Asn Phe Thr Phe Ser Tyr Thr Phe
Glu 405 410 415Asp Val Pro
Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp Arg 420
425 430Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu
Tyr Tyr Leu Ser Arg Thr 435 440
445Asn Thr Pro Ser Gly Thr Thr Thr Gln Ser Arg Leu Gln Phe Ser Gln 450
455 460Ala Gly Ala Ser Asp Ile Arg Asp
Gln Ser Arg Asn Trp Leu Pro Gly465 470
475 480Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Ser
Ala Asp Asn Asn 485 490
495Asn Ser Glu Tyr Ser Trp Thr Gly Ala Thr Lys Tyr His Leu Asn Gly
500 505 510Arg Asp Ser Leu Val Asn
Pro Gly Pro Ala Met Ala Ser His Lys Asp 515 520
525Asp Glu Glu Lys Phe Phe Pro Gln Ser Gly Val Leu Ile Phe
Gly Lys 530 535 540Gln Gly Ser Glu Lys
Thr Asn Val Asp Ile Glu Lys Val Met Ile Thr545 550
555 560Asp Glu Glu Glu Ile Arg Thr Thr Asn Pro
Val Ala Thr Glu Gln Tyr 565 570
575Gly Ser Val Ser Thr Asn Leu Gln Arg Gly Asn Arg Gln Ala Ala Thr
580 585 590Ala Asp Val Asn Thr
Gln Gly Val Leu Pro Gly Met Val Trp Gln Asp 595
600 605Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys
Ile Pro His Thr 610 615 620Asp Gly His
Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu Lys625
630 635 640His Pro Pro Pro Gln Ile Leu
Ile Lys Asn Thr Pro Val Pro Ala Asn 645
650 655Pro Ser Thr Thr Phe Ser Ala Ala Lys Phe Ala Ser
Phe Ile Thr Gln 660 665 670Tyr
Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln Lys 675
680 685Glu Asn Ser Lys Arg Trp Asn Pro Glu
Ile Gln Tyr Thr Ser Asn Tyr 690 695
700Asn Lys Ser Val Asn Val Asp Phe Thr Val Asp Thr Asn Gly Val Tyr705
710 715 720Ser Glu Pro Arg
Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu 725
730 73598597PRTAdeno-associated virus - 2 98Ala Pro
Gly Lys Lys Arg Pro Val Glu His Ser Pro Val Glu Pro Asp1 5
10 15Ser Ser Ser Gly Thr Gly Lys Ala
Gly Gln Gln Pro Ala Arg Lys Arg 20 25
30Leu Asn Phe Gly Gln Thr Gly Asp Ala Asp Ser Val Pro Asp Pro
Gln 35 40 45Pro Leu Gly Gln Pro
Pro Ala Ala Pro Ser Gly Leu Gly Thr Asn Thr 50 55
60Met Ala Thr Gly Ser Gly Ala Pro Met Ala Asp Asn Asn Glu
Gly Ala65 70 75 80Asp
Gly Val Gly Asn Ser Ser Gly Asn Trp His Cys Asp Ser Thr Trp
85 90 95Met Gly Asp Arg Val Ile Thr
Thr Ser Thr Arg Thr Trp Ala Leu Pro 100 105
110Thr Tyr Asn Asn His Leu Tyr Lys Gln Ile Ser Ser Gln Ser
Gly Ala 115 120 125Ser Asn Asp Asn
His Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe 130
135 140Asp Phe Asn Arg Phe His Cys His Phe Ser Pro Arg
Asp Trp Gln Arg145 150 155
160Leu Ile Asn Asn Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys
165 170 175Leu Phe Asn Ile Gln
Val Lys Glu Val Thr Gln Asn Asp Gly Thr Thr 180
185 190Thr Ile Ala Asn Asn Leu Thr Ser Thr Val Gln Val
Phe Thr Asp Ser 195 200 205Glu Tyr
Gln Leu Pro Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu 210
215 220Pro Pro Phe Pro Ala Asp Val Phe Met Val Pro
Gln Tyr Gly Tyr Leu225 230 235
240Thr Leu Asn Asn Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys
245 250 255Leu Glu Tyr Phe
Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Thr 260
265 270Phe Ser Tyr Thr Phe Glu Asp Val Pro Phe His
Ser Ser Tyr Ala His 275 280 285Ser
Gln Ser Leu Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu 290
295 300Tyr Tyr Leu Ser Arg Thr Asn Thr Pro Ser
Gly Thr Thr Thr Gln Ser305 310 315
320Arg Leu Gln Phe Ser Gln Ala Gly Ala Ser Asp Ile Arg Asp Gln
Ser 325 330 335Arg Asn Trp
Leu Pro Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys 340
345 350Thr Ser Ala Asp Asn Asn Asn Ser Glu Tyr
Ser Trp Thr Gly Ala Thr 355 360
365Lys Tyr His Leu Asn Gly Arg Asp Ser Leu Val Asn Pro Gly Pro Ala 370
375 380Met Ala Ser His Lys Asp Asp Glu
Glu Lys Phe Phe Pro Gln Ser Gly385 390
395 400Val Leu Ile Phe Gly Lys Gln Gly Ser Glu Lys Thr
Asn Val Asp Ile 405 410
415Glu Lys Val Met Ile Thr Asp Glu Glu Glu Ile Arg Thr Thr Asn Pro
420 425 430Val Ala Thr Glu Gln Tyr
Gly Ser Val Ser Thr Asn Leu Gln Arg Gly 435 440
445Asn Arg Gln Ala Ala Thr Ala Asp Val Asn Thr Gln Gly Val
Leu Pro 450 455 460Gly Met Val Trp Gln
Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp465 470
475 480Ala Lys Ile Pro His Thr Asp Gly His Phe
His Pro Ser Pro Leu Met 485 490
495Gly Gly Phe Gly Leu Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn
500 505 510Thr Pro Val Pro Ala
Asn Pro Ser Thr Thr Phe Ser Ala Ala Lys Phe 515
520 525Ala Ser Phe Ile Thr Gln Tyr Ser Thr Gly Gln Val
Ser Val Glu Ile 530 535 540Glu Trp Glu
Leu Gln Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile545
550 555 560Gln Tyr Thr Ser Asn Tyr Asn
Lys Ser Val Asn Val Asp Phe Thr Val 565
570 575Asp Thr Asn Gly Val Tyr Ser Glu Pro Arg Pro Ile
Gly Thr Arg Tyr 580 585 590Leu
Thr Arg Asn Leu 59599533PRTAdeno-associated virus - 2 99Met Ala
Thr Gly Ser Gly Ala Pro Met Ala Asp Asn Asn Glu Gly Ala1 5
10 15Asp Gly Val Gly Asn Ser Ser Gly
Asn Trp His Cys Asp Ser Thr Trp 20 25
30Met Gly Asp Arg Val Ile Thr Thr Ser Thr Arg Thr Trp Ala Leu
Pro 35 40 45Thr Tyr Asn Asn His
Leu Tyr Lys Gln Ile Ser Ser Gln Ser Gly Ala 50 55
60Ser Asn Asp Asn His Tyr Phe Gly Tyr Ser Thr Pro Trp Gly
Tyr Phe65 70 75 80Asp
Phe Asn Arg Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg
85 90 95Leu Ile Asn Asn Asn Trp Gly
Phe Arg Pro Lys Arg Leu Asn Phe Lys 100 105
110Leu Phe Asn Ile Gln Val Lys Glu Val Thr Gln Asn Asp Gly
Thr Thr 115 120 125Thr Ile Ala Asn
Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser 130
135 140Glu Tyr Gln Leu Pro Tyr Val Leu Gly Ser Ala His
Gln Gly Cys Leu145 150 155
160Pro Pro Phe Pro Ala Asp Val Phe Met Val Pro Gln Tyr Gly Tyr Leu
165 170 175Thr Leu Asn Asn Gly
Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys 180
185 190Leu Glu Tyr Phe Pro Ser Gln Met Leu Arg Thr Gly
Asn Asn Phe Thr 195 200 205Phe Ser
Tyr Thr Phe Glu Asp Val Pro Phe His Ser Ser Tyr Ala His 210
215 220Ser Gln Ser Leu Asp Arg Leu Met Asn Pro Leu
Ile Asp Gln Tyr Leu225 230 235
240Tyr Tyr Leu Ser Arg Thr Asn Thr Pro Ser Gly Thr Thr Thr Gln Ser
245 250 255Arg Leu Gln Phe
Ser Gln Ala Gly Ala Ser Asp Ile Arg Asp Gln Ser 260
265 270Arg Asn Trp Leu Pro Gly Pro Cys Tyr Arg Gln
Gln Arg Val Ser Lys 275 280 285Thr
Ser Ala Asp Asn Asn Asn Ser Glu Tyr Ser Trp Thr Gly Ala Thr 290
295 300Lys Tyr His Leu Asn Gly Arg Asp Ser Leu
Val Asn Pro Gly Pro Ala305 310 315
320Met Ala Ser His Lys Asp Asp Glu Glu Lys Phe Phe Pro Gln Ser
Gly 325 330 335Val Leu Ile
Phe Gly Lys Gln Gly Ser Glu Lys Thr Asn Val Asp Ile 340
345 350Glu Lys Val Met Ile Thr Asp Glu Glu Glu
Ile Arg Thr Thr Asn Pro 355 360
365Val Ala Thr Glu Gln Tyr Gly Ser Val Ser Thr Asn Leu Gln Arg Gly 370
375 380Asn Arg Gln Ala Ala Thr Ala Asp
Val Asn Thr Gln Gly Val Leu Pro385 390
395 400Gly Met Val Trp Gln Asp Arg Asp Val Tyr Leu Gln
Gly Pro Ile Trp 405 410
415Ala Lys Ile Pro His Thr Asp Gly His Phe His Pro Ser Pro Leu Met
420 425 430Gly Gly Phe Gly Leu Lys
His Pro Pro Pro Gln Ile Leu Ile Lys Asn 435 440
445Thr Pro Val Pro Ala Asn Pro Ser Thr Thr Phe Ser Ala Ala
Lys Phe 450 455 460Ala Ser Phe Ile Thr
Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile465 470
475 480Glu Trp Glu Leu Gln Lys Glu Asn Ser Lys
Arg Trp Asn Pro Glu Ile 485 490
495Gln Tyr Thr Ser Asn Tyr Asn Lys Ser Val Asn Val Asp Phe Thr Val
500 505 510Asp Thr Asn Gly Val
Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr 515
520 525Leu Thr Arg Asn Leu 530100143RNAArtificial
SequenceDescription of Artificial Sequence Synthetic polynucleotide
100guuuuagagc uaggccaaca ugaggaucac ccaugucugc agggccuagc aaguuaaaau
60aaggcuaguc cguuaucaac uuggccaaca ugaggaucac ccaugucugc agggccaagu
120ggcaccgagu cggugcuuuu uuu
143101199RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 101guuuuagagc uaggccaaca ugaggaucac
ccaugucugc agggccuagc aaguuaaaau 60aaggcuaguc cguuaucaac uuggccaaca
ugaggaucac ccaugucugc agggccaagu 120ggcaccgagu cggugcggga gcacaugagg
aucacccaug ugcgacuccc acagucacug 180gggagucuuc ccuuuuuuu
199102149RNAArtificial
SequenceDescription of Artificial Sequence Synthetic polynucleotide
102guuuaagagc uaugcuggaa acagcauagc aaguuuaaau aaggcuaguc cguuaucaac
60uugaaaaagu ggcaccgagu cggugcggga gcacaugagg aucacccaug ugcgacuccc
120acagucacug gggagucuuc ccuuuuuuu
149103186RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 103uucuagauca ucgaaacaug aggaucaccc
auaucugcag ucgacaucga aacaugagga 60ucacccaugu cugcagucga caucgaaaca
ugaggaucac ccaugucugc agucgacauc 120gaaacaugag gaucacccau gucugcaguc
gacaucgaaa ucgauaagcu ucagaucaga 180uccuag
18610419RNAArtificial
SequenceDescription of Artificial Sequence Synthetic oligonucleotide
104acaugaggau cacccaugu
1910519RNAArtificial SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 105acaugaggau cacccauau
1910614RNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 106ccacagucac uggg
1410789RNAArtificial
SequenceDescription of Artificial Sequence Synthetic oligonucleotide
107acaugaggau cacccauguc ugcagggccu agcaaguuaa aauaaggcua guccguuauc
60aacuuggcca acaugaggau cacccaugu
89108141RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 108guuuuagagc uaggccggag cagacgauau
ggcgucgcuc cggccuagca aguuaaaaua 60aggcuagucc guuaucaacu uggccggagc
agacgauaug gcgucgcucc ggccaagugg 120caccgagucg gugcuuuuuu u
14110932RNAArtificial
SequenceDescription of Artificial Sequence Synthetic oligonucleotide
109gccggagcag acgauauggc gucgcuccgg cc
32110112RNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 110guuuaagagc uaugcuggaa acagcauagc
aaguuuaaau aaggcuaguc cguuaucaac 60uugaaaaagu ggcaccgagu cggugccuga
augccugcga gcaucuuuuu uu 11211119RNAArtificial
SequenceDescription of Artificial Sequence Synthetic oligonucleotide
111cugaaugccu gcgagcauc
1911218DNAArtificial SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 112gaagaagctg caggaggt
1811318DNAArtificial SequenceDescription of
Artificial Sequence Synthetic oligonucleotide 113gctggagggg aagtggtc
1811436DNAArtificial
SequenceDescription of Artificial Sequence Synthetic oligonucleotide
114gaagaagctg caggaggtgc tggaggggaa gtggtc
36115346DNAArtificial SequenceDescription of Artificial Sequence
Synthetic polynucleotide 115tgaagaagct gcaggaggtg ctggagggga
agtggtccgg atcttgaaga agctgcagga 60ggtgctggag gggaagtggt ccggatcttg
aagaagctgc aggaggtgct ggaggggaag 120tggtccggat cttgaagaag ctgcaggagg
tgctggaggg gaagtggtcc ggatcttgaa 180gaagctgcag gaggtgctgg aggggaagtg
gtccggatct tgaagaagct gcaggaggtg 240ctggagggga agtggtccgg atcttgaaga
agctgcagga ggtgctggag gggaagtggt 300ccggatcttg aagaagctgc aggaggtgct
ggaggggaag tggtcc 3461169DNAArtificial
SequenceDescription of Artificial Sequence Synthetic oligonucleotide
116gtagatgga
911720DNAArtificial SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 117cggcgtagcc gatgtcgcgc
201189DNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 118aagggttca
91199DNAArtificial
SequenceDescription of Artificial Sequence Synthetic oligonucleotide
119gcgtgggcg
912018DNAArtificial SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 120aagggttcag cgtgggcg
1812119DNAArtificial SequenceDescription of Artificial
Sequence Synthetic oligonucleotide 121aagggttcag gcgtgggcg
1912220DNAArtificial
SequenceDescription of Artificial Sequence Synthetic oligonucleotide
122aagggttcag tgcgtgggcg
2012326DNAArtificial SequenceDescription of Artificial Sequence Synthetic
oligonucleotide 123gatccgccac aacatcgagg acggca
26
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