Patent application title: Method of Measuring Adaptive Immunity
Inventors:
Harlan S. Robins (Seattle, WA, US)
Harlan S. Robins (Seattle, WA, US)
Edus H. Warren, Iii (Bainbridge Island, WA, US)
Christopher Scott Carlson (Kirkland, WA, US)
Christopher Scott Carlson (Kirkland, WA, US)
Assignees:
Fred Hutchinson Cancer Research Center
IPC8 Class: AC12Q168FI
USPC Class:
506 2
Class name: Combinatorial chemistry technology: method, library, apparatus method specially adapted for identifying a library member
Publication date: 2014-09-11
Patent application number: 20140256567
Abstract:
A method of measuring immunocompetence is described. This method provides
a means for assessing the effects of diseases or conditions that
compromise the immune system and of therapies aimed to reconstitute it.
This method is based on quantifying T-cell diversity by calculating the
number of diverse T-cell receptor (TCR) beta chain variable regions from
blood cells.Claims:
1. A method for monitoring a disease state of an individual by
determining one or more correlating clonotypes of the disease wherein the
disease comprises an autoimmune disease, an infectious disease or cancer,
the method comprising: (a) generating one or more clonotype profiles by
sequencing individual, spatially isolated nucleic acid molecules, i.
wherein each spatially isolated nucleic acid molecule comprises a
recombined sequence from a T-cell and/or B-cell from at least one sample
from the individual, ii. wherein each of said samples comprises T-cells
and/or B-cells and wherein the at least one sample is from a tissue
affected by the disease, and iii. wherein each of the one or more
clonotype profiles comprises at least 1000 sequence reads of at least 30
base pairs per read; (b) determining one or more correlating clonotypes
of the disease in the individual using the one or more clonotype profiles
from the tissue affected by the disease; (c) generating one or more
clonotype profiles of one or more blood samples of the individual,
wherein each of the one or more clonotype profiles of the one or more
blood samples comprises at least 1000 sequence reads of at least 30 base
pairs per read; and (d) monitoring a level of the one or more correlating
clonotypes in the one or more blood samples of the individual by
comparing the one or more correlating clonotypes identified in step (b)
with the one or more clonotype profiles generated in step (c).
2. The method of claim 1, wherein the determining one or more correlating clonotypes of the disease in the individual is achieved by comparing the clonotype profile from the tissue affected by the disease with one or more other clonotype profiles.
3. The method of claim 1, wherein the determining one or more correlating clonotypes of the disease in the individual is achieved by identifying high level clonotypes in the clonotype profile from the tissue affected by the disease.
4. The method of claim 1, wherein said disease is an autoimmune disease and said one or more correlating clonotypes are present in a peak state of the disease, wherein the peak state of the disease is a flare state of the autoimmune disease.
5. The method of claim 1, wherein said T-cells and/or B-cells comprise a subset of T-cells and/or B cells.
6. The method of claim 5, wherein said subset of T-cells and/or B-cells are enriched by interaction with a marker.
7. The method of claim 6, wherein said marker is a cell surface marker on the subset of T-cells and/or B-cells.
8. The method of claim 5, wherein said subset of T-cells and/or B-cells interact with an antigen specifically present in the disease.
9. The method of claim 1, wherein the disease is systemic lupus erythematosus or multiple sclerosis.
10. The method of claim 1, wherein said nucleic acid molecules are amplified.
11. The method of claim 10, wherein each of said samples comprises at least 10,000 B-cells.
12. The method of claim 10, wherein each of said samples comprises at least 10,000 T-cells.
13. The method of claim 1, wherein each of said samples comprises at least 10,000 B-cells and wherein each of said clonotype profiles comprises at least 100,000 sequence reads.
14. The method of claim 13, wherein each of said sequence reads comprises a paired-end read having an error rate of one percent or more.
15. The method of claim 1, wherein each of said samples comprises at least 10,000 T-cells and wherein each of said clonotype profiles comprises at least 100,000 sequence reads.
16. The method of claim 15, wherein each of said sequence reads comprises a paired-end read having an error rate of one percent or more.
17. A method for monitoring a disease of an individual by quantifying a diversity of an immune receptor repertoire of said individual, wherein said disease comprises an autoimmune disease, an infectious disease, or cancer, said method comprising: (a) quantifying diversity of one or more immune receptor repertoires by sequencing by synthesis using an array of spatially isolated, individual DNA molecules, (i) wherein each spatially isolated, individual DNA molecule comprises a rearranged sequence from a T-cell and/or a B-cell from at least one sample obtained from said individual, (ii) wherein each of said samples comprises T cells and/or B cells and wherein said at least one sample is obtained from a tissue affected by said disease, and (iii) wherein said diversity of said one or more immune receptor repertoires comprises at least 10.sup.4 unique rearranged sequences of at least 30 base pairs per read; (b) determining one or more measurements of diversity of said one or more immune receptor repertoires of said disease in said individual using said one or more quantified diversities determined from said tissue affected by said disease; (c) determining one or more measurements of diversity of one or more immune receptor repertoires from blood samples obtained from said individual, wherein each of said measurements of diversity of said blood samples comprises at least 10.sup.4 sequence reads of at least 30 base pairs per read; and (d) monitoring a change in diversity in said one or more measurements of diversity from said blood samples of said individual by comparing said one or more measurements of diversity determined in step (b) with said one or more measurements of diversity determined in step (c).
18. The method of claim 17, wherein said determining one or more measurements of diversity of said immune receptor repertoire of said disease in said individual comprises comparing a measurement of diversity measured from a sample comprising tissue affected by said disease with one or more other measurements of diversity.
19. The method of claim 17, wherein said disease is an autoimmune disease and wherein said one or more measurements of diversity are measured before a patient undergoes treatment.
20. The method of claim 17, wherein said T-cells and/or B-cells comprise a subset of T-cells and/or B-cells.
21. The method of claim 20, wherein said subset of T-cells and/or B-cells comprise a marker.
22. The method of claim 21, wherein said marker is a cell surface marker on the subset of T-cells and/or B-cells.
23. The method of claim 20, wherein said subset of T-cells and/or B-cells interact with an antigen specifically present in the disease.
24. The method of claim 17, wherein said disease is an autoimmune disease.
25. The method of claim 17, wherein said individual DNA molecules are amplified.
26. The method of claim 25, wherein each of said samples comprises at least 10,000 B cells.
27. The method of claim 25, wherein each of said samples comprises at least 10,000 T cells.
28. The method of claim 17, wherein each of said samples comprises at least 10,000 B cells, and wherein each of said measurements of diversity comprises at least 100,000 sequence reads.
29. The method of claim 28, wherein each of said sequence reads comprise a read having an error rate of one percent or more.
30. The method of claim 17, wherein each of said samples comprises at least 10,000 T-cells and wherein each of said measurements of diversity comprises at least 100,000 sequence reads.
31. The method of claim 30, wherein each of said sequence reads comprise a read having an error rate of one percent or more.
Description:
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. application Ser. No. 13/960,761, filed on Aug. 6, 2013, which is a continuation of U.S. application Ser. No. 12/794,507, filed on Jun. 4, 2010, which claims the benefit of U.S. Provisional Application No. 61/220,344, filed on Jun. 25, 2009, which are all herein incorporated by reference in their entireties.
TECHNICAL FIELD
[0002] What is described is a method to measure the adaptive immunity of a patient by analyzing the diversity of T cell receptor genes or antibody genes using large scale sequencing of nucleic acid extracted from adaptive immune system cells.
BACKGROUND
[0003] Immunocompetence is the ability of the body to produce a normal immune response (i.e., antibody production and/or cell-mediated immunity) following exposure to a pathogen, which might be a live organism (such as a bacterium or fungus), a virus, or specific antigenic components isolated from a pathogen and introduced in a vaccine. Immunocompetence is the opposite of immunodeficiency or immuno-incompetent or immunocompromised. Several examples would be a newborn that does not yet have a fully functioning immune system but may have maternally transmitted antibody (immunodeficient); a late stage AIDS patient with a failed or failing immune system (immuno-incompetent); a transplant recipient taking medication so their body will not reject the donated organ (immunocompromised); age-related attenuation of T cell function in the elderly; or individuals exposed to radiation or chemotherapeutic drugs. There may be cases of overlap but these terms are all indicators of a dysfunctional immune system. In reference to lymphocytes, immunocompetence means that a B cell or T cell is mature and can recognize antigens and allow a person to mount an immune response.
[0004] Immunocompetence depends on the ability of the adaptive immune system to mount an immune response specific for any potential foreign antigens, using the highly polymorphic receptors encoded by B cells (immunoglobulins, Igs) and T cells (T cell receptors, TCRs).
[0005] Igs expressed by B cells are proteins consisting of four polypeptide chains, two heavy chains (H chains) and two light chains (L chains), forming an H2L2 structure. Each pair of H and L chains contains a hypervariable domain, consisting of a VL and a VH region, and a constant domain. The H chains of Igs are of several types, μ, δ, γ, α, and β. The diversity of Igs within an individual is mainly determined by the hypervariable domain. The V domain of H chains is created by the combinatorial joining of three types of germline gene segments, the VH, DH, and JH segments. Hypervariable domain sequence diversity is further increased by independent addition and deletion of nucleotides at the VH-DH, DH-JH, and VH-JH junctions during the process of Ig gene rearrangement. In this respect, immunocompetence is reflected in the diversity of Igs.
[0006] TCRs expressed by αβ T cells are proteins consisting of two transmembrane polypeptide chains (α and β), expressed from the TCRA and TCRB genes, respectively. Similar TCR proteins are expressed in gamma-delta T cells, from the TCRD and TCRG loci. Each TCR peptide contains variable complementarity determining regions (CDRs), as well as framework regions (FRs) and a constant region. The sequence diversity of αβ T cells is largely determined by the amino acid sequence of the third complementarity-determining region (CDR3) loops of the α and β chain variable domains, which diversity is a result of recombination between variable (V.sub.β), diversity (D.sub.β), and joining (J.sub.β) gene segments in the β chain locus, and between analogous V.sub.α and J.sub.α gene segments in the α chain locus, respectively. The existence of multiple such gene segments in the TCR α and β chain loci allows for a large number of distinct CDR3 sequences to be encoded. CDR3 sequence diversity is further increased by independent addition and deletion of nucleotides at the V.sub.β-D.sub.β, D.sub.β-J.sub.β, and V.sub.α-J.sub.α junctions during the process of TCR gene rearrangement. In this respect, immunocompetence is reflected in the diversity of TCRs.
[0007] There exists a long-felt need for methods of assessing or measuring the adaptive immune system of patients in a variety of settings, whether immunocompetence in the immunocompromised, or dysregulated adaptive immunity in autoimmune disease. A demand exists for methods of diagnosing a disease state or the effects of aging by assessing the immunocompetence of a patient. In the same way results of therapies that modify the immune system need to be monitored by assessing the immunocompetence of the patient while undergoing the treatment. Conversely, a demand exists for methods to monitor the adaptive immune system in the context of autoimmune disease flares and remissions, in order to monitor response to therapy, or the need to initiate prophylactic therapy pre-symptomatically.
SUMMARY
[0008] One aspect of the invention is composition comprising:
[0009] a multiplicity of V-segment primers, wherein each primer comprises a sequence that is complementary to a single functional V segment or a small family of V segments; and
[0010] a multiplicity of J-segment primers, wherein each primer comprises a sequence that is complementary to a J segment; wherein the V segment and J-segment primers permit amplification of a TCR CDR3 region by a multiplex polymerase chain reaction (PCR) to produce a multiplicity of amplified DNA molecules sufficient to quantify the diversity of the TCR genes. One embodiment of the invention is the composition, wherein each V-segment primer comprises a sequence that is complementary to a single Vβ segment, and each J segment primer comprises a sequence that is complementary to a Jβ segment, and wherein V segment and J-segment primers permit amplification of a TCRβ CDR3 region. Another embodiment is the composition, wherein each V-segment primer comprises a sequence that is complementary to a single functional Vα segment, and each J segment primer comprises a sequence that is complementary to a Jα segment, and wherein V segment and J-segment primers permit amplification of a TCRα CDR3 region.
[0011] Another embodiment of the invention is the composition, wherein the V segment primers hybridize with a conserved segment, and have similar annealing strength. Another embodiment is wherein the V segment primer is anchored at position -43 in the Vβ segment relative to the recombination signal sequence (RSS). Another embodiment is wherein the multiplicity of V segment primers consist of at least 45 primers specific to 45 different Vβ genes. Another embodiment is wherein the V segment primers have sequences that are selected from the group consisting of SEQ ID NOS:1-45. Another embodiment is wherein the V segment primers have sequences that are selected from the group consisting of SEQ ID NOS:58-102. Another embodiment is wherein there is a V segment primer for each Vβ segment.
[0012] Another embodiment of the invention is the composition, wherein the J segment primers hybridize with a conserved framework region element of the JP segment, and have similar annealing strength. The composition of claim 2, wherein the multiplicity of J segment primers consist of at least thirteen primers specific to thirteen different JP genes. Another embodiment is The composition of claim 2, wherein the J segment primers have sequences that are selected from the group consisting of SEQ ID NOS:46-57. Another embodiment is wherein the J segment primers have sequences that are selected from the group consisting of SEQ ID NOS:102-113. Another embodiment is wherein there is a J segment primer for each JP segment. Another embodiment is wherein all J segment primers anneal to the same conserved motif.
[0013] Another embodiment of the invention is the composition, wherein the amplified DNA molecule starts from said conserved motif and amplifies adequate sequence to diagnostically identify the J segment and includes the CDR3 junction and extends into the V segment. Another embodiment is wherein the amplified JP gene segments each have a unique four base tag at positions +11 through +14 downstream of the RSS site. Another aspect of the invention is the composition further comprising a set of sequencing oligonucleotides, wherein the sequencing oligonucleotides hybridize to a regions within the amplified DNA molecules. An embodiment is wherein the sequencing oligonucleotides hybridize adjacent to a four base tag within the amplified JP gene segments at positions +11 through +14 downstream of the RSS site. Another embodiment is wherein the sequencing oligonucleotides are selected from the group consisting of SEG ID NOS:58-70. Another embodiment is wherein the V-segment or J-segment are selected to contain a sequence error-correction by merger of closely related sequences. Another embodiment is the composition, further comprising a universal C segment primer for generating cDNA from mRNA.
[0014] Another aspect of the invention is a composition comprising:
[0015] a multiplicity of V segment primers, wherein each V segment primer comprises a sequence that is complementary to a single functional V segment or a small family of V segments; and
[0016] a multiplicity of J segment primers, wherein each J segment primer comprises a sequence that is complementary to a J segment;
[0017] wherein the V segment and J segment primers permit amplification of the TCRG CDR3 region by a multiplex polymerase chain reaction (PCR) to produce a multiplicity of amplified DNA molecules sufficient to quantify the diversity of antibody heavy chain genes.
[0018] Another aspect of the invention is a composition comprising:
[0019] a multiplicity of V segment primers, wherein each V segment primer comprises a sequence that is complementary to a single functional V segment or a small family of V segments; and
[0020] a multiplicity of J segment primers, wherein each J segment primer comprises a sequence that is complementary to a J segment;
[0021] wherein the V segment and J segment primers permit amplification of antibody heavy chain (IGH) CDR3 region by a multiplex polymerase chain reaction (PCR) to produce a multiplicity of amplified DNA molecules sufficient to quantify the diversity of antibody heavy chain genes.
[0022] Another aspect of the invention is a composition comprising:
[0023] a multiplicity of V segment primers, wherein each V segment primer comprises a sequence that is complementary to a single functional V segment or a small family of V segments; and
[0024] a multiplicity of J segment primers, wherein each J segment primer comprises a sequence that is complementary to a J segment;
[0025] wherein the V segment and J segment primers permit amplification of antibody light chain (IGL) VL region by a multiplex polymerase chain reaction (PCR) to produce a multiplicity of amplified DNA molecules sufficient to quantify the diversity of antibody light chain genes.
[0026] Another aspect of the invention is a method comprising:
[0027] selecting a multiplicity of V segment primers, wherein each V segment primer comprises a sequence that is complementary to a single functional V segment or a small family of V segments; and
[0028] selecting a multiplicity of J segment primers, wherein each J segment primer comprises a sequence that is complementary to a J segment;
[0029] combining the V segment and J segment primers with a sample of genomic DNA to permit amplification of a CDR3 region by a multiplex polymerase chain reaction (PCR) to produce a multiplicity of amplified DNA molecules sufficient to quantify the diversity of the TCR genes.
[0030] One embodiment of the invention is the method wherein each V segment primer comprises a sequence that is complementary to a single functional Vβ segment, and each J segment primer comprises a sequence that is complementary to a Jβ segment; and wherein combining the V segment and J segment primers with a sample of genomic DNA permits amplification of a TCR CDR3 region by a multiplex polymerase chain reaction (PCR) and produces a multiplicity of amplified DNA molecules. Another embodiment is wherein each V segment primer comprises a sequence that is complementary to a single functional Vα segment, and each J segment primer comprises a sequence that is complementary to a Jα segment; and wherein combining the V segment and J segment primers with a sample of genomic DNA permits amplification of a TCR CDR3 region by a multiplex polymerase chain reaction (PCR) and produces a multiplicity of amplified DNA molecules.
[0031] Another embodiment of the invention is the method further comprising a step of sequencing the amplified DNA molecules. Another embodiment is wherein the sequencing step utilizes a set of sequencing oligonucleotides, that hybridize to regions within the amplified DNA molecules. Another embodiment is the method, further comprising a step of calculating the total diversity of TCRβ CDR3 sequences among the amplified DNA molecules. Another embodiment is wherein the method shows that the total diversity of a normal human subject is greater than 1*106 sequences, greater than 2*106 sequences, or greater than 3*106 sequences.
[0032] Another aspect of the invention is a method of diagnosing immunodeficiency in a human patient, comprising measuring the diversity of TCR CDR3 sequences of the patient, and comparing the diversity of the subject to the diversity obtained from a normal subject. An embodiment of the invention is the method, wherein measuring the diversity of TCR sequences comprises the steps of:
[0033] selecting a multiplicity of V segment primers, wherein each V segment primer comprises a sequence that is complementary to a single functional V segment or a small family of V segments; and
[0034] selecting a multiplicity of J segment primers, wherein each J segment primer comprises a sequence that is complementary to a J segment;
[0035] combining the V segment and J segment primers with a sample of genomic DNA to permit amplification of a TCR CDR3 region by a multiplex polymerase chain reaction (PCR) to produce a multiplicity of amplified DNA molecules;
[0036] sequencing the amplified DNA molecules;
[0037] calculating the total diversity of TCR CDR3 sequences among the amplified DNA molecules.
[0038] An embodiment of the invention is the method, wherein comparing the diversity is determined by calculating using the following equation:
Δ ( t ) = x E ( n x ) measurement 1 + 2 - x E ( n x ) measurement 2 = S ∫ 0 ∞ - λ ( 1 - - λ t ) G ( λ ) ##EQU00001##
[0039] wherein G(λ) is the empirical distribution function of the parameters λ1, . . . , ZS, nx is the number of clonotypes sequenced exactly x times, and
E ( n x ) = S ∫ 0 ∞ ( - λ λ x x ! ) G ( λ ) . ##EQU00002##
[0040] Another embodiment of the invention is the method, wherein the diversity of at least two samples of genomic DNA are compared. Another embodiment is wherein one sample of genomic DNA is from a patient and the other sample is from a normal subject. Another embodiment is wherein one sample of genomic DNA is from a patient before a therapeutic treatment and the other sample is from the patient after treatment. Another embodiment is wherein the two samples of genomic DNA are from the same patient at different times during treatment. Another embodiment is wherein a disease is diagnosed based on the comparison of diversity among the samples of genomic DNA. Another embodiment is wherein the immunocompetence of a human patient is assessed by the comparison.
DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS
[0041] The TCR and Ig genes can generate millions of distinct proteins via somatic mutation. Because of this diversity-generating mechanism, the hypervariable complementarity determining regions of these genes can encode sequences that can interact with millions of ligands, and these regions are linked to a constant region that can transmit a signal to the cell indicating binding of the protein's cognate ligand.
[0042] The adaptive immune system employs several strategies to generate a repertoire of T- and B-cell antigen receptors with sufficient diversity to recognize the universe of potential pathogens. In αβ and γδ T cells, which primarily recognize peptide antigens presented by MHC molecules, most of this receptor diversity is contained within the third complementarity-determining region (CDR3) of the T cell receptor (TCR) α and β chains (or γ and δ chains). Although it has been estimated that the adaptive immune system can generate up to 1018 distinct TCR αβ pairs, direct experimental assessment of TCR CDR3 diversity has not been possible.
[0043] What is described herein is a novel method of measuring TCR CDR3 diversity that is based on single molecule DNA sequencing, and use this approach to sequence the CDR3 regions in millions of rearranged TCRβ genes isolated from peripheral blood T cells of two healthy adults.
[0044] The ability of the adaptive immune system to mount an immune response specific for any of the vast number of potential foreign antigens to which an individual might be exposed relies on the highly polymorphic receptors encoded by B cells (immunoglobulins) and T cells (T cell receptors; TCRs). The TCRs expressed by αβ T cells, which primarily recognize peptide antigens presented by major histocompatibility complex (MHC) class I and II molecules, are heterodimeric proteins consisting of two transmembrane polypeptide chains (α and β), each containing one variable and one constant domain. The peptide specificity of αβ T cells is in large part determined by the amino acid sequence encoded in the third complementarity-determining region (CDR3) loops of the α and β chain variable domains. The CDR3 regions of the β and α chains are formed by recombination between noncontiguous variable (V.sub.β), diversity (D.sub.β), and joining (J.sub.β) gene segments in the β chain locus, and between analogous V.sub.α and J.sub.α gene segments in the α chain locus, respectively. The existence of multiple such gene segments in the TCR α and β chain loci allows for a large number of distinct CDR3 sequences to be encoded. CDR3 sequence diversity is further increased by template-independent addition and deletion of nucleotides at the V.sub.β-D.sub.β, D.sub.β-J.sub.β, and V.sub.α-J.sub.α junctions during the process of TCR gene rearrangement.
[0045] Previous attempts to assess the diversity of receptors in the adult human αβ T cell repertoire relied on examining rearranged TCR α and β chain genes expressed in small, well-defined subsets of the repertoire, followed by extrapolation of the diversity present in these subsets to the entire repertoire, to estimate approximately 106 unique TCRβ chain CDR3 sequences per individual, with 10-20% of these unique TCRβ CDR3 sequences expressed by cells in the antigen-experienced CD45RO+ compartment. The accuracy and precision of this estimate is severely limited by the need to extrapolate the diversity observed in hundreds of sequences to the entire repertoire, and it is possible that the actual number of unique TCRβ chain CDR3 sequences in the αβ T cell repertoire is significantly larger than 1×106.
[0046] Recent advances in high-throughput DNA sequencing technology have made possible significantly deeper sequencing than capillary-based technologies. A complex library of template molecules carrying universal PCR adapter sequences at each end is hybridized to a lawn of complementary oligonucleotides immobilized on a solid surface. Solid phase PCR is utilized to amplify the hybridized library, resulting in millions of template clusters on the surface, each comprising multiple (1,000) identical copies of a single DNA molecule from the original library. A 30-54 bp interval in the molecules in each cluster is sequenced using reversible dye-termination chemistry, to permit simultaneous sequencing from genomic DNA of the rearranged TCRβ chain CDR3 regions carried in millions of T cells. This approach enables direct sequencing of a significant fraction of the uniquely rearranged TCRβ CDR3 regions in populations of αβ T cells, which thereby permits estimation of the relative frequency of each CDR3 sequence in the population.
[0047] Accurate estimation of the diversity of TCRβ CDR3 sequences in the entire αβ T cell repertoire from the diversity measured in a finite sample of T cells requires an estimate of the number of CDR3 sequences present in the repertoire that were not observed in the sample. TCRβ chain CDR3 diversity in the entire αβ T cell repertoire were estimated using direct measurements of the number of unique TCRβ CDR3 sequences observed in blood samples containing millions of αβ T cells. The results herein identify a lower bound for TCRβ CDR3 diversity in the CD4+ and CD8+ T cell compartments that is several fold higher than previous estimates. In addition, the results herein demonstrat that there are at least 1.5×106 unique TCRβ CDR3 sequences in the CD45RO+ compartment of antigen-experienced T-cells, a large proportion of which are present at low relative frequency. The existence of such a diverse population of TCRβ CDR3 sequences in antigen-experienced cells has not been previously demonstrated.
[0048] The diverse pool of TCRβ chains in each healthy individual is a sample from an estimated theoretical space of greater than 1011 possible sequences. However, the realized set of rearranged of TCRs is not evenly sampled from this theoretical space. Different Vβ's and Jβ's are found with over a thousand-fold frequency difference. Additionally, the insertion rates of nucleotides are strongly biased. This reduced space of realized TCRβ sequences leads to the possibility of shared β chains between people. With the sequence data generated by the methods described herein, the in vivo J usage, V usage, mono- and di-nucleotide biases, and position dependent amino acid usage can be computed. These biases significantly narrow the size of the sequence space from which TCRβ are selected, suggesting that different individuals share TCRβ chains with identical amino acid sequences. Results herein show that many thousands of such identical sequences are shared pairwise between individual human genomes.
[0049] The assay technology uses two pools of primers to provide for a highly multiplexed PCR reaction. The "forward" pool has a primer specific to each V segment in the gene (several primers targeting a highly conserved region are used, to simultaneously capture many V segments). The "reverse" pool primers anneal to a conserved sequence in the joining ("J") segment. The amplified segment pool includes adequate sequence to identify each J segment and also to allow for a J-segment-specific primer to anneal for resequencing. This enables direct observation of a large fraction of the somatic rearrangements present in an individual. This in turn enables rapid comparison of the TCR repertoire in individuals with an autoimmune disorder (or other target disease indication) against the TCR repertoire of controls.
[0050] The adaptive immune system can in theory generate an enormous diversity of T cell receptor CDR3 sequences--far more than are likely to be expressed in any one individual at any one time. Previous attempts to measure what fraction of this theoretical diversity is actually utilized in the adult αβ T cell repertoire, however, have not permitted accurate assessment of the diversity. What is described herein is the development of a novel approach to this question that is based on single molecule DNA sequencing and an analytic computational approach to estimation of repertoire diversity using diversity measurements in finite samples. The analysis demonstrated that the number of unique TCRβ CDR3 sequences in the adult repertoire significantly exceeds previous estimates based on exhaustive capillary sequencing of small segments of the repertoire. The TCRβ chain diversity in the CD45ROpopulation (enriched for naive T cells) observed using the methods described herein is five-fold larger than previously reported. A major discovery is the number of unique TCRβ CDR3 sequences expressed in antigen-experienced CD45RO+ T cells--the results herein show that this number is between 10 and 20 times larger than expected based on previous results of others. The frequency distribution of CDR3 sequences in CD45RO+ cells suggests that the T cell repertoire contains a large number of clones with a small clone size.
[0051] The results herein show that the realized set of TCRβ chains are sampled non-uniformly from the huge potential space of sequences. In particular, the β chains sequences closer to germ line (few insertions and deletions at the V-D and D-J boundaries) appear to be created at a relatively high frequency. TCR sequences close to germ line are shared between different people because the germ line sequence for the V's, D's, and J's are shared, modulo a small number of polymorphisms, among the human population.
[0052] The T cell receptors expressed by mature αβ T cells are heterodimers whose two constituent chains are generated by independent rearrangement events of the TCR α and β chain variable loci. The α chain has less diversity than the β chain, so a higher fraction of α's are shared between individuals, and hundreds of exact TCR αβ receptors are shared between any pair of individuals.
Cells
[0053] B cells and T cells can be obtained from a variety of tissue samples including marrow, thymus, lymph glands, peripheral tissues and blood, but peripheral blood is most easily accessed. Peripheral blood samples are obtained by phlebotomy from subjects. Peripheral blood mononuclear cells (PBMC) are isolated by techniques known to those of skill in the art, e.g., by Ficoll-Hypaque® density gradient separation. Preferably, whole PBMCs are used for analysis. The B and/or T lymphocytes, instead, may be flow sorted into multiple compartments for each subject: e.g. CD8+CD45RO+/- and CD4+CD45RO+/- using fluorescently labeled anti-human antibodies, e.g, CD4 FITC (clone M-T466, Miltenyi Biotec), CD8 PE (clone RPA-T8, BD Biosciences), CD45RO ECD (clone UCHL-1, Beckman Coulter), and CD45RO APC (clone UCHL-1, BD Biosciences). Staining of total PBMCs may be done with the appropriate combination of antibodies, followed by washing cells before analysis. Lymphocyte subsets can be isolated by FACS sorting, e.g., by a BD FACSAria® cell-sorting system (BD Biosciences) and by analyzing results with FlowJo software (Treestar Inc.), and also by conceptually similar methods involving specific antibodies immobilized to surfaces or beads.
Nucleic Acid Extraction
[0054] Total genomic DNA is extracted from cells, e.g., by using the QIAamp® DNA blood Mini Kit (QIAGEN®). The approximate mass of a single haploid genome is 3 μg. Preferably, at least 100,000 to 200,000 cells are used for analysis of diversity, i.e., about 0.6 to 1.2 μg DNA from diploid T cells. Using PBMCs as a source, the number of T cells can be estimated to be about 30% of total cells.
[0055] Alternatively, total nucleic acid can be isolated from cells, including both genomic DNA and mRNA. If diversity is to be measured from mRNA in the nucleic acid extract, the mRNA must be converted to cDNA prior to measurement. This can readily be done by methods of one of ordinary skill.
DNA Amplification
[0056] A multiplex PCR system is used to amplify rearranged TCR loci from genomic DNA, preferably from a CDR3 region, more preferably from a TCRα, TCRγ or TCRδ CDR3 region, most preferably from a TCRβ CDR3 region.
[0057] In general, a multiplex PCR system may use at least 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25, preferably 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, or 39, most preferably 40, 41, 42, 43, 44, or 45 forward primers, in which each forward primer is specific to a sequence corresponding to one or more TRB V region segments shown in SEQ ID NOS:114-248; and at least 3, 4, 5, 6, or 7, preferably 8, 9, 10, 11, 12 or 13 reverse primers, in which each reverse primer is specific to a sequence corresponding to one or more TRB J region segments shown in SEQ ID NOS:249-261. Most preferably, there is a J segment primer for every J segment.
[0058] Preferably, the primers are designed not to cross an intron/exon boundary. The forward primers must preferably anneal to the V segments in a region of relatively strong sequence conservation between V segments so as to maximize the conservation of sequence among these primers. Accordingly, this minimizes the potential for differential annealing properties of each primer, and so that the amplified region between V and J primers contains sufficient TCR V sequence information to identify the specific V gene segment used.
[0059] Preferably, the J segment primers hybridize with a conserved element of the J segment, and have similar annealing strength. Most preferably, all J segment primers anneal to the same conserved framework region motif. The forward and reverse primers are both preferably modified at the 5' end with the universal forward primer sequence compatible with a DNA sequencer.
[0060] For example, a multiplex PCR system may use 45 forward primers (Table 1), each specific to a functional TCR Vβ segment, and thirteen reverse primers (Table 2), each specific to a TCR Jβ segment. Xn and Yn correspond to polynucleotides of lengths n and m, respectively, which would be specific to the single molecule sequencing technology being used to read out the assay.
TABLE-US-00001 TABLE 1 TCR-Vβ Forward primer sequences SEQ TRBV gene ID segment(s) NO: Primer sequence* TRBV2 1 XnTCAAATTTCACTCTGAAGATCCGGTC CACAA TRBV3-1 2 XnGCTCACTTAAATCTTCACATCAATTC CCTGG TRBV4-1 3 XnCTTAAACCTTCACCTACACGCCCTGC TRBV(4-2, 4-3) 4 XnCTTATTCCTTCACCTACACACCCTGC TRBV5-1 5 XnGCTCTGAGATGAATGTGAGCACCTTG TRBV5-3 6 XnGCTCTGAGATGAATGTGAGTGCCTTG TRBV(5-4, 5-5, 7 XnGCTCTGAGCTGAATGTGAACGCCTTG 5-6, 5-7, 5-8) TRBV6-1 8 XnTCGCTCAGGCTGGAGTCGGCTG TRBV(6-2, 6-3) 9 XnGCTGGGGTTGGAGTCGGCTG TRBV6-4 10 XnCCCTCACGTTGGCGTCTGCTG TRBV6-5 11 XnGCTCAGGCTGCTGTCGGCTG TRBV6-6 12 XnCGCTCAGGCTGGAGTTGGCTG TRBV6-7 13 XnCCCCTCAAGCTGGAGTCAGCTG TRBV6-8 14 XnCACTCAGGCTGGTGTCGGCTG TRBV6-9 15 XnCGCTCAGGCTGGAGTCAGCTG TRBV7-1 16 XnCCACTCTGAAGTTCCAGCGCACAC TRBV7-2 17 XnCACTCTGACGATCCAGCGCACAC TRBV7-3 18 XnCTCTACTCTGAAGATCCAGCGCACAG TRBV7-4 19 XnCCACTCTGAAGATCCAGCGCACAG TRBV7-6 20 XnCACTCTGACGATCCAGCGCACAG TRBV7-7 21 XnCCACTCTGACGATTCAGCGCACAG TRBV7-8 22 XnCCACTCTGAAGATCCAGCGCACAC TRBV7-9 23 XnCACCTTGGAGATCCAGCGCACAG TRBV9 24 XnGCACTCTGAACTAAACCTGAGCTCTC TG TRBV10-1 25 XnCCCCTCACTCTGGAGTCTGCTG TRBV10-2 26 XnCCCCCTCACTCTGGAGTCAGCTA TRBV10-3 27 XnCCTCCTCACTCTGGAGTCCGCTA TRBV(11-1, 11-3) 28 XnCCACTCTCAAGATCCAGCCTGCAG TRBV11-2 29 XnCTCCACTCTCAAGATCCAGCCTGCAA TRBV(12-3, 12-4, 30 XnCCACTCTGAAGATCCAGCCCTCAG 12-5) TRBV13 31 XnCATTCTGAACTGAACATGAGCTCCTT GG TRBV14 32 XnCTACTCTGAAGGTGCAGCCTGCAG TRBV15 33 XnGATAACTTCCAATCCAGGAGGCCGAA CA TRBV16 34 XnCTGTAGCCTTGAGATCCAGGCTACGA TRBV17 35 XnCTTCCACGCTGAAGATCCATCCCG TRBV18 36 XnGCATCCTGAGGATCCAGCAGGTAG TRBV19 37 XnCCTCTCACTGTGACATCGGCCC TRBV20-1 38 XnCTTGTCCACTCTGACAGTGACCAGTG TRBV23-1 39 XnCAGCCTGGCAATCCTGTCCTCAG TRBV24-1 40 XnCTCCCTGTCCCTAGAGTCTGCCAT TRBV25-1 41 XnCCCTGACCCTGGAGTCTGCCA TRBV27 42 XnCCCTGATCCTGGAGTCGCCCA TRBV28 43 XnCTCCCTGATTCTGGAGTCCGCCA TRBV29-1 44 XnCTAACATTCTCAACTCTGACTGTGAG CAACA TRBV30 45 XnCGGCAGTTCATCCTGAGTTCTAAGAA GC
TABLE-US-00002 TABLE 2 TCR-Jβ Reverse Primer Sequences TRBJ gene SEQ ID segment NO: Primer sequence* TRBJ1-1 46 YmTTACCTACAACTGTGAGTCTGGTGCCTTGTC CAAA TRBJ1-2 47 YmACCTACAACGGTTAACCTGGTCCCCGAACCG AA TRBJ1-3 48 YmACCTACAACAGTGAGCCAACTTCCCTCTCCA AA TRBJ1-4 49 YmCCAAGACAGAGAGCTGGGTTCCACTGCCAAA TRBJ1-5 483 YmACCTAGGATGGAGAGTCGAGTCCCATCACCA AA TRBJ1-6 50 YmCTGTCACAGTGAGCCTGGTCCCGTTCCCAAA TRBJ2-1 51 YmCGGTGAGCCGTGTCCCTGGCCCGAA TRBJ2-2 52 YmCCAGTACGGTCAGCCTAGAGCCTTCTCCAAA TRBJ2-3 53 YmACTGTCAGCCGGGTGCCTGGGCCAAA TRBJ2-4 54 YmAGAGCCGGGTCCCGGCGCCGAA TRBJ2-5 55 YmGGAGCCGCGTGCCTGGCCCGAA TRBJ2-6 56 YmGTCAGCCTGCTGCCGGCCCCGAA TRBJ2-7 57 YmGTGAGCCTGGTGCCCGGCCCGAA
[0061] The 45 forward PCR primers of Table 1 are complementary to each of the 48 functional Variable segments, and the thirteen reverse PCR primers of Table 2 are complementary to each of the functional joining (J) gene segments from the TRB locus (TRBJ). The TRB V region segments are identified in the Sequence Listing at SEQ ID NOS:114-248 and the TRB J region segments are at SEQ ID NOS:249-261. The primers have been designed such that adequate information is present within the amplified sequence to identify both the V and J genes uniquely (>40 base pairs of sequence upstream of the V gene recombination signal sequence (RSS), and >30 base pairs downstream of the J gene RSS). Alternative primers may be selected by one of ordinary skill from the V and J regions of the genes of each TCR subunit.
[0062] The forward primers are modified at the 5' end with the universal forward primer sequence compatible with the DNA sequencer (Xn of Table 1). Similarly, all of the reverse primers are modified with a universal reverse primer sequence (Ym of Table 2). One example of such universal primers is shown in Tables 3 and 4, for the Illumina GAII single-end read sequencing system. The 45 TCR Vβ forward primers anneal to the Vβ segments in a region of relatively strong sequence conservation between Vβ segments so as to maximize the conservation of sequence among these primers.
TABLE-US-00003 TABLE 3 TCR-Vβ Forward primer sequences SEQ TRBV gene ID segment(s) NO: Primer sequence* TRBV2 58 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTTCAAATTTCACTCTGAAGATCCGGTCCACAA TRBV3-1 59 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTGCTCACTTAAATCTTCACATCAATTCCCT- GG TRBV4-1 60 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCTTAAACCTTCACCTACACGCCCTGC TRBV(4-2, 4-3) 61 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCTTATTCCTTCACCTACACACCCTGC TRBV5-1 62 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTGCTCTGAGATGAATGTGAGCACCTTG TRBV5-3 63 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTGCTCTGAGATGAATGTGAGTGCCTTG TRBV(5-4, 5-5, 64 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTGCTCTGAGCTGAATGTGAACGCCTTG 5-6,5-7,5-8) TRBV6-1 65 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTTCGCTCAGGCTGGAGTCGGCTG TRBV(6-2, 6-3) 66 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTGCTGGGGTTGGAGTCGGCTG TRBV6-4 67 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCCCTCACGTTGGCGTCTGCTG TRBV6-5 68 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTGCTCAGGCTGCTGTCGGCTG TRBV6-6 69 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCGCTCAGGCTGGAGTTGGCTG TRBV6-7 70 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCCCCTCAAGCTGGAGTCAGCTG TRBV6-8 71 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCACTCAGGCTGGTGTCGGCTG TRBV6-9 72 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCGCTCAGGCTGGAGTCAGCTG TRBV7-1 73 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCCACTCTGAAGTTCCAGCGCACAC TRBV7-2 74 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCACTCTGACGATCCAGCGCACAC TRBV7-3 75 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCTCTACTCTGAAGATCCAGCGCACAG TRBV7-4 76 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCCACTCTGAAGATCCAGCGCACAG TRBV7-6 77 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCACTCTGACGATCCAGCGCACAG TRBV7-7 78 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCCACTCTGACGATTCAGCGCACAG TRBV7-8 79 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCCACTCTGAAGATCCAGCGCACAC TRBV7-9 80 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCACCTTGGAGATCCAGCGCACAG TRBV9 81 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTGCACTCTGAACTAAACCTGAGCTCTCTG TRBV10-1 82 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCCCCTCACTCTGGAGTCTGCTG TRBV10-2 83 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCCCCCTCACTCTGGAGTCAGCTA TRBV10-3 84 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCCTCCTCACTCTGGAGTCCGCTA TRBV(11-1,11-3) 85 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCCACTCTCAAGATCCAGCCTGCAG TRBV11-2 86 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCTCCACTCTCAAGATCCAGCCTGCAA TRBV(12-3, 87 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCCACTCTGAAGATCCAGCCCTCAG 12-4, 12-5) TRBV13 88 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCATTCTGAACTGAACATGAGCTCCTTGG TRBV14 89 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCTACTCTGAAGGTGCAGCCTGCAG TRBV15 90 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTGATAACTTCCAATCCAGGAGGCCGAACA TRBV16 91 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCTGTAGCCTTGAGATCCAGGCTACGA TRBV17 92 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCTTCCACGCTGAAGATCCATCCCG TRBV18 93 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTGCATCCTGAGGATCCAGCAGGTAG TRBV19 94 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCCTCTCACTGTGACATCGGCCC TRBV20-1 95 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCTTGTCCACTCTGACAGTGACCAGTG TRBV23-1 96 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCAGCCTGGCAATCCTGTCCTCAG TRBV24-1 97 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCTCCCTGTCCCTAGAGTCTGCCAT TRBV25-1 98 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCCCTGACCCTGGAGTCTGCCA TRBV27 99 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCCCTGATCCTGGAGTCGCCCA TRBV28 100 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCTCCCTGATTCTGGAGTCCGCCA TRBV29-1 101 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCTAACATTCTCAACTCTGACTGTGAGCAACA TRBV30 102 CAAGCAGAAGACGGCATACGAGCTCTTCCGATCTCGGCAGTTCATCCTGAGTTCTAAGAAGC
TABLE-US-00004 TABLE 4 TCR-Jβ Reverse Primer Sequences TRBJ gene SEQ ID segment NO: Primer sequence* TRBJ1-1 103 AATGATACGGCGACCACCGAGATCTTTACCTACAACTGTGAGTCTGGTGCCTTGTCCAAA TRBJ1-2 468 AATGATACGGCGACCACCGAGATCTACCTACAACGGTTAACCTGGTCCCCGAACCGAA TRBJ1-3 104 AATGATACGGCGACCACCGAGATCTACCTACAACAGTGAGCCAACTTCCCTCTCCAAA TRBJ1-4 105 AATGATACGGCGACCACCGAGATCTCCAAGACAGAGAGCTGGGTTCCACTGCCAAA TRBJ1-5 484 AATGATACGGCGACCACCGAGATCTACCTAGGATGGAGAGTCGAGTCCCATCACCAAA TRBJ1-6 106 AATGATACGGCGACCACCGAGATCTCTGTCACAGTGAGCCTGGTCCCGTTCCCAAA TRBJ2-1 107 AATGATACGGCGACCACCGAGATCTCGGTGAGCCGTGTCCCTGGCCCGAA TRBJ2-2 108 AATGATACGGCGACCACCGAGATCTCCAGTACGGTCAGCCTAGAGCCTTCTCCAAA TRBJ2-3 109 AATGATACGGCGACCACCGAGATCTACTGTCAGCCGGGTGCCTGGGCCAAA TRBJ2-4 110 AATGATACGGCGACCACCGAGATCTAGAGCCGGGTCCCGGCGCCGAA TRBJ2-5 111 AATGATACGGCGACCACCGAGATCTGGAGCCGCGTGCCTGGCCCGAA TRBJ2-6 112 AATGATACGGCGACCACCGAGATCTGTCAGCCTGCTGCCGGCCCCGAA TRBJ2-7 113 AATGATACGGCGACCACCGAGATCTGTGAGCCTGGTGCCCGGCCCGAA *bold sequence indicates universal R oligonucleotide for the sequence analysis
[0063] The total PCR product for a rearranged TCRβ CDR3 region using this system is expected to be approximately 200 bp long. Genomic templates are PCR amplified using a pool of the 45 TCR Vβ F primers (the "VF pool") and a pool of the twelve TCR Jβ R primers (the "JR pool"). For example, 50 μl PCR reactions may be used with 1.0 μM VF pool (22 nM for each unique TCR Vβ F primer), 1.0 μM R pool (77 nM for each unique TCRBJR primer), 1×QIAGEN Multiple PCR master mix (QIAGEN part number 206145), 10% Q-solution (QIAGEN), and 16 ng/ul gDNA.
[0064] The IGH primer set was designed to try to accommodate the potential for somatic hypermutation within the rearranged IGH genes, as is observed after initial stimulation of naive B cells. Consequently all primers were designed to be slightly longer than normal, and to anchor the 3' ends of each primer into highly conserved sequences of three or more nucleotides that should be resistant to both functional and non-functional somatic mutations.
[0065] The IGHJ reverse primers were designed to anchor the 3' end of each PCR primer on a highly conserved GGGG sequence motif within the IGHJ segments. These sequences are shown in Table 5. Underlined sequence are ten base pairs in from RSS that may be deleted. These were excluded from barcode design. Bold sequence is the reverse complement of the IGH J reverse PCR primers. Italicized sequence is the barcode for J identity (eight barcodes reveal six genes, and two alleles within genes). Further sequence within underlined segment may reveal additional allelic identities.
TABLE-US-00005 TABLE 5 SEQ ID IgH J segment NO: Sequence >IGHJ4*01/1-48 452 ACTACTTTGACTACTGGGGCCAAGGAACCCTGGTCACCGTCTCCTCAG >IGHJ4*03/1-48 453 GCTACTTTGACTACTGGGGCCAAGGGACCCTGGTCACCGTCTCCTCAG >IGHJ4*02/1-48 454 ACTACTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAG >IGHJ3*01/1-50 455 TGATGCTTTTGATGTCTGGGGCCAAGGGACAATGGTCACCGTCTCTTCAG >IGHJ3*02/1-50 456 TGATGCTTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTCTCTTCAG >IGHJ6*01/1-63 457 ATTACTACTACTACTACGGTATGGACGTCTGGGGGCAAGGGACCACGGTCACCGTCTCCTCAG >IGHJ6*02/1-62 458 ATTACTACTACTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCAG >IGHJ6*04/1-63 459 ATTACTACTACTACTACGGTATGGACGTCTGGGGCAAAGGGACCACGGTCACCGTCTCCTCAG >IGHJ6*03/1-62 460 ATTACTACTACTACTACTACATGGACGTCTGGGGCAAAGGGACCACGGTCACCGTCTCCTCAG >IGHJ2*01/1-53 461 CTACTGGTACTTCGATCTCTGGGGCCGTGGCACCCTGGTCACTGTCTCCTCAG >IGHJ5*01/1-51 462 ACAACTGGTTCGACTCCTGGGGCCAAGGAACCCTGGTCACCGTCTCCTCAG >IGHJ5*02/1-51 463 ACAACTGGTTCGACCCCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAG >IGHJ1*01/1-52 464 GCTGAATACTTCCAGCACTGGGGCCAGGGCACCCTGGTCACCGTCTCCTCAG >IGHJ2P*01/1-61 465 CTACAAGTGCTTGGAGCACTGGGGCAGGGCAGCCCGGACACCGTCTCCCTGGGAACGTCAG >IGHJ1P*01/1-54 466 AAAGGTGCTGGGGGTCCCCTGAACCCGACCCGCCCTGAGACCGCAGCCACATCA >IGHJ3P*01/1-52 467 CTTGCGGTTGGACTTCCCAGCCGACAGTGGTGGTCTGGCTTCTGAGGGGTCA
[0066] Sequences of the IGHJ reverse PCR primers are shown in Table 6.
TABLE-US-00006 TABLE 6 IgH J SEQ ID segment NO: sequence >IGHJ4_1 421 TGAGGAGACGGTGACCAGGGTTCCTTGGCCC >IGHJ4_3 422 TGAGGAGACGGTGACCAGGGTCCCTTGGCCC >IGHJ4_2 423 TGAGGAGACGGTGACCAGGGTTCCCTGGCCC >IGHJ3_12 424 CTGAAGAGACGGTGACCATTGTCCCTTGGCCC >IGHJ6_1 425 CTGAGGAGACGGTGACCGTGGTCCCTTGCCCC >IGHJ6_2 426 TGAGGAGACGGTGACCGTGGTCCCTTGGCCC >IGHJ6_34 427 CTGAGGAGACGGTGACCGTGGTCCCTTTGCCC >IGHJ2_1 428 CTGAGGAGACAGTGACCAGGGTGCCACGGCCC >IGHJ5_1 429 CTGAGGAGACGGTGACCAGGGTTCCTTGGCCC >IGHJ5_2 430 CTGAGGAGACGGTGACCAGGGTTCCCTGGCCC >IGHJ1_1 431 CTGAGGAGACGGTGACCAGGGTGCCCTGGCCC
[0067] V primers were designed in a conserved in region of FR2 between the two conserved tryptophan (W) codons.
[0068] The primer sequences are anchored at the 3' end on a tryptophan codon for all IGHV families that conserve this codon. his allows for the last three nucleotides (tryptophan's TGG) to anchor on sequence that is expected to be resistant to somatic hypermutation, providing a 3' anchor of five out of six nucleotides for each primer. The upstream sequence is extended further than normal, and includes degenerate nucleotides to allow for mismatches induced by hypermutation (or between closely relate IGH V families) without dramatically changing the annealing characteristics of the primer, as shown in Table 7. The sequences of the V gene segments are SEQ ID NOS:262-420.
TABLE-US-00007 TABLE 7 IgH V SEQ ID segment NO: sequence >IGHV1 443 TGGGTGCACCAGGTCCANGNACAAGGGCTT GAGTGG >IGHV2 444 TGGGTGCGACAGGCTCGNGNACAACGCCTT GAGTGG >IGHV3 445 TGGGTGCGCCAGATGCCNGNGAAAGGCCTG GAGTGG >IGHV4 446 TGGGTCCGCCAGSCYCCNGNGAAGGGGCTG GAGTGG >IGHV5 447 TGGGTCCGCCAGGCTCCNGNAAAGGGGCTG GAGTGG >IGHV6 448 TGGGTCTGCCAGGCTCCNGNGAAGGGGCAG GAGTGG >IGH7_3.25p 449 TGTGTCCGCCAGGCTCCAGGGAATGGGCTG GAGTTGG >IGH8_3.54p 450 TCAGATTCCCAAGCTCCAGGGAAGGGGCTG GAGTGAG >IGH9_3.63p 451 TGGGTCAATGAGACTCTAGGGAAGGGGCTG GAGGGAG
[0069] Thermal cycling conditions may follow methods of those skilled in the art. For example, using a PCR Express thermal cycler (Hybaid, Ashford, UK), the following cycling conditions may be used: 1 cycle at 95° C. for 15 minutes, 25 to 40 cycles at 94° C. for 30 seconds, 59° C. for 30 seconds and 72° C. for 1 minute, followed by one cycle at 72° C. for 10 minutes.
Sequencing
[0070] Sequencing is achieved using a set of sequencing oligonucleotides that hybridize to a defined region within the amplified DNA molecules.
[0071] Preferably, the amplified J gene segments each have a unique four base tag at positions +11 through +14 downstream from the RSS site. Accordingly, the sequencing oligonucleotides hybridize adjacent to a four base tag within the amplified Jβ gene segments at positions +11 through +14 downstream of the RSS site.
[0072] For example, sequencing oligonucleotides for TCRB may be designed to anneal to a consensus nucleotide motif observed just downstream of this "tag", so that the first four bases of a sequence read will uniquely identify the J segment (Table 8).
TABLE-US-00008 TABLE 8 Sequencing oligonucleotides Sequencing SEQ ID oligonucleotide NO: Oligonucleotide sequence Jseq 1-1 470 ACAACTGTGAGTCTGGTGCCTTGT CCAAAGAAA Jseq 1-2 471 ACAACGGTTAACCTGGTCCCCGAA CCGAAGGTG Jseq 1-3 472 ACAACAGTGAGCCAACTTCCCTCT CCAAAATAT Jseq 1-4 473 AAGACAGAGAGCTGGGTTCCACTG CCAAAAAAC Jseq 1-5 474 AGGATGGAGAGTCGAGTCCCATCA CCAAAATGC Jseq 1-6 475 GTCACAGTGAGCCTGGTCCCGTTC CCAAAGTGG Jseq 2-1 476 AGCACGGTGAGCCGTGTCCCTGGC CCGAAGAAC Jseq 2-2 477 AGTACGGTCAGCCTAGAGCCTTCT CCAAAAAAC Jseq 2-3 478 AGCACTGTCAGCCGGGTGCCTGGG CCAAAATAC Jseq 2-4 479 AGCACTGAGAGCCGGGTCCCGGCG CCGAAGTAC Jseq 2-5 480 AGCACCAGGAGCCGCGTGCCTGGC CCGAAGTAC Jseq 2-6 481 AGCACGGTCAGCCTGCTGCCGGCC CCGAAAGTC Jseq 2-7 482 GTGACCGTGAGCCTGGTGCCCGGC CCGAAGTAC
[0073] The information used to assign the J and V segment of a sequence read is entirely contained within the amplified sequence, and does not rely upon the identity of the PCR primers. These sequencing oligonucleotides were selected such that promiscuous priming of a sequencing reaction for one J segment by an oligonucleotide specific to another J segment would generate sequence data starting at exactly the same nucleotide as sequence data from the correct sequencing oligonucleotide. In this way, promiscuous annealing of the sequencing oligonucleotides did not impact the quality of the sequence data generated.
[0074] The average length of the CDR3 region, defined as the nucleotides between the second conserved cysteine of the V segment and the conserved phenylalanine of the J segment, is 35+/-3, so sequences starting from the Jβ segment tag will nearly always capture the complete V-D-J junction in a 50 base pair read.
[0075] TCR βJ gene segments are roughly 50 base pair in length. PCR primers that anneal and extend to mismatched sequences are referred to as promiscuous primers. The TCR Jβ Reverse PCR primers were designed to minimize overlap with the sequencing oligonucleotides to minimize promiscuous priming in the context of multiplex PCR. The 13 TCR Jβ reverse primers are anchored at the 3' end on the consensus splice site motif, with minimal overlap of the sequencing primers. The TCR Jβ primers provide consistent annealing temperature using the sequencer program under default parameters.
[0076] For the sequencing reaction, the IGHJ sequencing primers extend three nucleotides across the conserved CAG sequences as shown in Table 9.
TABLE-US-00009 TABLE 9 IgH J SEQ ID segment NO: sequence >IGHJSEQ4_1 432 TGAGGAGACGGTGACCAGGGTTCCTTG GCCCCAG >IGHJSEQ4_3 433 TGAGGAGACGGTGACCAGGGTCCCTTG GCCCCAG >IGHJSEQ4_2 434 TGAGGAGACGGTGACCAGGGTTCCCTG GCCCCAG >IGHJSEQ3_12 435 CTGAAGAGACGGTGACCATTGTCCCTT GGCCCCAG >IGHJSEQ6_1 436 CTGAGGAGACGGTGACCGTGGTCCCTT GCCCCCAG >IGHJSEQ6_2 437 TGAGGAGACGGTGACCGTGGTCCCTTG GCCCCAG >IGHJSEQ6_34 438 CTGAGGAGACGGTGACCGTGGTCCCTT TGCCCCAG >IGHJSEQ2_1 439 CTGAGGAGACAGTGACCAGGGTGCCAC GGCCCCAG >IGHJSEQ5_1 440 CTGAGGAGACGGTGACCAGGGTTCCTT GGCCCCAG >IGHJSEQ5_2 441 CTGAGGAGACGGTGACCAGGGTTCCCT GGCCCCAG >IGHJSEQ1_1 442 CTGAGGAGACGGTGACCAGGGTGCCCT GGCCCCAG
Processing Sequence Data
[0077] For rapid analysis of sequencing results, an algorithm can be developed by one of ordinary skill. A preferred method is as follows.
[0078] The use of a PCR step to amplify the TCRβ CDR3 regions prior to sequencing could potentially introduce a systematic bias in the inferred relative abundance of the sequences, due to differences in the efficiency of PCR amplification of CDR3 regions utilizing different Vβ and Jβ gene segments. Each cycle of PCR amplification potentially introduces a bias of average magnitude 1.51/15=1.027. Thus, the 25 cycles of PCR introduces a total bias of average magnitude 1.02725=1.95 in the inferred relative abundance of distinct CDR3 region sequences.
[0079] Sequenced reads were filtered for those including CDR3 sequences. Sequencer data processing involves a series of steps to remove errors in the primary sequence of each read, and to compress the data. A complexity filter removes approximately 20% of the sequences that are misreads from the sequencer. Then, sequences were required to have a minimum of a six base match to both one of the thirteen TCRB J-regions and one of 54 V-regions. Applying the filter to the control lane containing phage sequence, on average only one sequence in 7-8 million passed these steps. Finally, a nearest neighbor algorithm was used to collapse the data into unique sequences by merging closely related sequences, in order to remove both PCR error and sequencing error.
[0080] Analyzing the data, the ratio of sequences in the PCR product must be derived working backward from the sequence data before estimating the true distribution of clonotypes in the blood. For each sequence observed a given number of times in the data herein, the probability that that sequence was sampled from a particular size PCR pool is estimated. Because the CDR3 regions sequenced are sampled randomly from a massive pool of PCR products, the number of observations for each sequence are drawn from Poisson distributions. The Poisson parameters are quantized according to the number of T cell genomes that provided the template for PCR. A simple Poisson mixture model both estimates these parameters and places a pairwise probability for each sequence being drawn from each distribution. This is an expectation maximization method which reconstructs the abundances of each sequence that was drawn from the blood.
[0081] To estimate diversity, the "unseen species" formula is employed. To apply this formula, unique adaptive immune receptors (e.g. TCRB) clonotypes takes the place of species. The mathematical solution provides that for a total number of TCRβ "species" or clonotypes, S, a sequencing experiment observes xs copies of sequence s. For all of the unobserved clonotypes, xs equals 0, and each TCR clonotype is "captured" in a blood draw according to a Poisson process with parameter λs. The number of T cell genomes sequenced in the first measurement 1, and in the second measurement. Since there are a large number of unique sequences, an integral will represent the sum. If G(λ) is the empirical distribution function of the parameters λ1, . . . , λs, and nx is the number of clonotypes sequenced exactly x times, then the total number of clonotypes, i.e., the measurement of diversity E, is given by the following formula:
E ( n x ) = S ∫ 0 ∞ ( - λ λ x x ! ) G ( λ ) . ##EQU00003##
[0082] For a given experiment, where T cells are sampled from some arbitrary source (e.g. a blood draw), the formula is used to estimate the total diversity of species in the entire source. The idea is that the sampled number of clonotypes at each size contains sufficient information to estimate the underlying distribution of clonotypes in the whole source. To derive the formula, the number of new species expected if the exact measurement was repeated was estimated. The limit of the formula as if repeating the measurements an infinite number of times. The result is the expect number of species in the total underlying source population. The value for Δ(t), the number of new clonotypes observed in a second measurement, should be determined, preferably using the following equation:
Δ ( t ) = x E ( n x ) msmt 1 + msmt 2 - x E ( n x ) msmt 1 = S ∫ 0 ∞ - λ ( 1 - e - λ t ) G ( λ ) ##EQU00004##
in which msmt1 and msmt2 are the number of clonotypes from measurement 1 and 2, respectively. Taylor expansion of 1-e.sup.-λt gives Δ(t)=E(x1)t-E(x2)t2+E(x3)t3- . . . , which can be approximated by replacing the expectations E(nx) with the observed numbers in the first measurement. Using in the numbers observed in the first measurement, this formula predicts that 1.6*105 new unique sequences should be observed in the second measurement. The actual value of the second measurement was 1.8*105 new TCRβ sequences, which implies that the prediction provided a valid lower bound on total diversity. An Euler's transformation was used to regularize Δ(t) to produce a lower bound for Δ(∞).
Using a Measurement of Diversity to Diagnose Disease
[0083] The measurement of diversity can be used to diagnose disease or the effects of a treatment, as follows. T cell and/or B cell receptor repertoires can be measured at various time points, e.g., after hematopoietic stem cell transplant (HSCT) treatment for leukemia. Both the change in diversity and the overall diversity of TCRB repertoire can be utilized to measure immunocompetence. A standard for the expected rate of immune reconstitution after transplant can be utilized. The rate of change in diversity between any two time points may be used to actively modify treatment. The overall diversity at a fixed time point is also an important measure, as this standard can be used to compare between different patients. In particular, the overall diversity is the measure that should correlate with the clinical definition of immune reconstitution. This information may be used to modify prophylactic drug regiments of antibiotics, antivirals, and antifungals, e.g., after HSCT.
[0084] The assessment of immune reconstitution after allogeneic hematopoietic cell transplantation can be determined by measuring changes in diversity. These techniques will also enhance the analysis of how lymphocyte diversity declines with age, as measured by analysis of T cell responses to vaccination. Further, the methods of the invention provide a means to evaluate investigational therapeutic agents (e.g., Interleukin-7 (IL-7)) that have a direct effect on the generation, growth, and development of αβ T cells. Moreover, application of these techniques to the study of thymic T cell populations will provide insight into the processes of both T cell receptor gene rearrangement as well as positive and negative selection of thymocytes.
[0085] A newborn that does not yet have a fully functioning immune system but may have maternally transmitted antibody is immunodeficient. A newborn is susceptible to a number of diseases until its immune system autonomously develops, and our measurement of the adaptive immune system may will likely prove useful with newborn patients.
[0086] Lymphocyte diversity can be assessed in other states of congenital or acquired immunodeficiency. An AIDS patient with a failed or failing immune system can be monitored to determine the stage of disease, and to measure a patient's response to therapies aimed to reconstitute immunocompetence.
[0087] Another application of the methods of the invention is to provide diagnostic measures for solid organ transplant recipients taking medication so their body will not reject the donated organ. Generally, these patients are under immunosuppressive therapies. Monitoring the immunocompetence of the host will assist before and after transplantation.
[0088] Individuals exposed to radiation or chemotherapeutic drugs are subject to bone marrow transplantations or otherwise require replenishment of T cell populations, along with associated immunocompetence. The methods of the invention provide a means for qualitatively and quantitatively assessing the bone marrow graft, or reconstitution of lymphocytes in the course of these treatments.
[0089] One manner of determining diversity is by comparing at least two samples of genomic DNA, preferably in which one sample of genomic DNA is from a patient and the other sample is from a normal subject, or alternatively, in which one sample of genomic DNA is from a patient before a therapeutic treatment and the other sample is from the patient after treatment, or in which the two samples of genomic DNA are from the same patient at different times during treatment. Another manner of diagnosis may be based on the comparison of diversity among the samples of genomic DNA, e.g., in which the immunocompetence of a human patient is assessed by the comparison.
Biomarkers
[0090] Shared TCR sequences between individuals represent a new class of potential biomarkers for a variety of diseases, including cancers, autoimmune diseases, and infectious diseases. These are the public T cells that have been reported for multiple human diseases. TCRs are useful as biomarkers because T cells are a result of clonal expansion, by which the immune system amplifies these biomarkers through rapid cell division. Following amplification, the TCRs are readily detected even if the target is small (e.g. an early stage tumor). TCRs are also useful as biomarkers because in many cases the T cells might additionally contribute to the disease causally and, therefore could constitute a drug target. T cells self interactions are thought to play a major role in several diseases associated with autoimmunity, e.g., multiple sclerosis, Type I diabetes, and rheumatoid arthritis.
EXAMPLES
Example 1
Sample Acquisition, PBMC Isolation, FACS Sorting and Genomic DNA Extraction
[0091] Peripheral blood samples from two healthy male donors aged 35 and 37 were obtained with written informed consent using forms approved by the Institutional Review Board of the Fred Hutchinson Cancer Research Center (FHCRC). Peripheral blood mononuclear cells (PBMC) were isolated by Ficoll-Hypaque® density gradient separation. The T-lymphocytes were flow sorted into four compartments for each subject: CD8+CD45RO+/- and CD4+CD45RO+/-. For the characterization of lymphocytes the following conjugated anti-human antibodies were used: CD4 FITC (clone M-T466, Miltenyi Biotec), CD8 PE (clone RPA-T8, BD Biosciences), CD45RO ECD (clone UCHL-1, Beckman Coulter), and CD45RO APC (clone UCHL-1, BD Biosciences). Staining of total PBMCs was done with the appropriate combination of antibodies for 20 minutes at 4° C., and stained cells were washed once before analysis. Lymphocyte subsets were isolated by FACS sorting in the BD FACSAria® cell-sorting system (BD Biosciences). Data were analyzed with FlowJo software (Treestar Inc.).
[0092] Total genomic DNA was extracted from sorted cells using the QIAamp® DNA blood Mini Kit (QIAGEN®). The approximate mass of a single haploid genome is 3 pg. In order to sample millions of rearranged TCRB in each T cell compartment, 6 to 27 micrograms of template DNA were obtained from each compartment (see Table 10).
TABLE-US-00010 TABLE 10 CD8+/ CD8+/ CD4+/ CD4+/ CD45RO- CD45RO+ CD45RO- CD45RO+ Donor cells (×106) 9.9 6.3 6.3 10 2 DNA (μg) 27 13 19 25 PCR cycles 25 25 30 30 clusters (K/tile) 29.3 27 102.3* 118.3* VJ sequences 3.0 2.0 4.4 4.2 (×106) Cells 4.9 4.8 3.3 9 1 DNA 12 13 6.6 19 PCR cycles 30 30 30 30 Clusters 116.3 121 119.5 124.6 VJ sequences 3.2 3.7 4.0 3.8 Cells NA NA NA 0.03 PCR DNA NA NA NA 0.015 Bias PCR cycles NA NA NA 25 + 15 assess- clusters NA NA NA 1.4/23.8 ment VJ sequences NA NA NA 1.6
Example 2
Virtual T Cell Receptor β Chain Spectratyping
[0093] Virtual TCR β chain spectratyping was performed as follows. Complementary DNA was synthesized from RNA extracted from sorted T cell populations and used as template for multiplex PCR amplification of the rearranged TCR β chain CDR3 region. Each multiplex reaction contained a 6-FAM-labeled antisense primer specific for the TCR β chain constant region, and two to five TCR β chain variable (TRBV) gene-specific sense primers. All 23 functional Vβ families were studied. PCR reactions were carried out on a Hybaid PCR Express thermal cycler (Hybaid, Ashford, UK) under the following cycling conditions: 1 cycle at 95° C. for 6 minutes, 40 cycles at 94° C. for 30 seconds, 58° C. for 30 seconds, and 72° C. for 40 seconds, followed by 1 cycle at 72° C. for 10 minutes. Each reaction contained cDNA template, 500 μM dNTPs, 2 mM MgCl2 and 1 unit of AmpliTaq Gold DNA polymerase (Perkin Elmer) in AmpliTaq Gold buffer, in a final volume of 20 μl. After completion, an aliquot of the PCR product was diluted 1:50 and analyzed using a DNA analyzer. The output of the DNA analyzer was converted to a distribution of fluorescence intensity vs. length by comparison with the fluorescence intensity trace of a reference sample containing known size standards.
Example 3
Multiplex PCR Amplification of TCRβ CDR3 Regions
[0094] The CDR3 junction region was defined operationally, as follows. The junction begins with the second conserved cysteine of the V-region and ends with the conserved phenylalanine of the J-region. Taking the reverse complements of the observed sequences and translating the flanking regions, the amino acids defining the junction boundaries were identified. The number of nucleotides between these boundaries determines the length and therefore the frame of the CDR3 region. In order to generate the template library for sequencing, a multiplex PCR system was selected to amplify rearranged TCRβ loci from genomic DNA. The multiplex PCR system uses 45 forward primers (Table 3), each specific to a functional TCR Vβ segment, and thirteen reverse primers (Table 4), each specific to a TCR Jβ segment. The primers were selected to provide that adequate information is present within the amplified sequence to identify both the V and J genes uniquely (>40 base pairs of sequence upstream of the V gene recombination signal sequence (RSS), and >30 base pairs downstream of the J gene RSS).
[0095] The forward primers are modified at the 5' end with the universal forward primer sequence compatible with the Illumina GA2 cluster station solid-phase PCR. Similarly, all of the reverse primers are modified with the GA2 universal reverse primer sequence. The 3' end of each forward primer is anchored at position -43 in the Vβ segment, relative to the recombination signal sequence (RSS), thereby providing a unique Vβ tag sequence within the amplified region. The thirteen reverse primers specific to each Jβ segment are anchored in the 3' intron, with the 3' end of each primer crossing the intron/exon junction. Thirteen sequencing primers complementary to the Jβ segments were designed that are complementary to the amplified portion of the Jβ segment, such that the first few bases of sequence generated will capture the unique Jβ tag sequence.
[0096] On average J deletions were 4 bp+/-2.5 bp, which implies that J deletions greater than 10 nucleotides occur in less than 1% of sequences. The thirteen different TCR JP gene segments each had a unique four base tag at positions +11 through +14 downstream of the RSS site. Thus, sequencing oligonucleotides were designed to anneal to a consensus nucleotide motif observed just downstream of this "tag", so that the first four bases of a sequence read will uniquely identify the J segment (Table 5).
[0097] The information used to assign the J and V segment of a sequence read is entirely contained within the amplified sequence, and does not rely upon the identity of the PCR primers. These sequencing oligonucleotides were selected such that promiscuous priming of a sequencing reaction for one J segment by an oligonucleotide specific to another J segment would generate sequence data starting at exactly the same nucleotide as sequence data from the correct sequencing oligonucleotide. In this way, promiscuous annealing of the sequencing oligonucleotides did not impact the quality of the sequence data generated.
[0098] The average length of the CDR3 region, defined following convention as the nucleotides between the second conserved cysteine of the V segment and the conserved phenylalanine of the J segment, is 35+/-3, so sequences starting from the Jβ segment tag will nearly always capture the complete VNDNJ junction in a 50 bp read.
[0099] TCR βJ gene segments are roughly 50 bp in length. PCR primers that anneal and extend to mismatched sequences are referred to as promiscuous primers. Because of the risk of promiscuous priming in the context of multiplex PCR, especially in the context of a gene family, the TCR Jβ Reverse PCR primers were designed to minimize overlap with the sequencing oligonucleotides. Thus, the 13 TCR Jβ reverse primers are anchored at the 3' end on the consensus splice site motif, with minimal overlap of the sequencing primers. The TCR Jβ primers were designed for a consistent annealing temperature (58 degrees in 50 mM salt) using the OligoCalc program under default parameters (http://www.basic.northwestern.edu/biotools/oligocalc.html).
[0100] The 45 TCR Vβ forward primers were designed to anneal to the Vβ segments in a region of relatively strong sequence conservation between Vβ segments, for two express purposes. First, maximizing the conservation of sequence among these primers minimizes the potential for differential annealing properties of each primer. Second, the primers were chosen such that the amplified region between V and J primers will contain sufficient TCR Vβ sequence information to identify the specific Vβ gene segment used. This obviates the risk of erroneous TCR Vβ gene segment assignment, in the event of promiscuous priming by the TCR Vβ primers. TCR Vβ forward primers were designed for all known non-pseudogenes in the TCRβ locus.
[0101] The total PCR product for a successfully rearranged TCRβ CDR3 region using this system is expected to be approximately 200 bp long. Genomic templates were PCR amplified using an equimolar pool of the 45 TCR Vβ F primers (the "VF pool") and an equimolar pool of the thirteen TCR Jβ R primers (the "JR pool"). 50 μl PCR reactions were set up at 1.0 μM VF pool (22 nM for each unique TCR Vβ F primer), 1.0 μM JR pool (77 nM for each unique TCRBJR primer), 1×QIAGEN Multiple PCR master mix (QIAGEN part number 206145), 10% Q-solution (QIAGEN), and 16 ng/ul gDNA. The following thermal cycling conditions were used in a PCR Express thermal cycler (Hybaid, Ashford, UK) under the following cycling conditions: 1 cycle at 95° C. for 15 minutes, 25 to 40 cycles at 94° C. for 30 seconds, 59° C. for 30 seconds and 72° C. for 1 minute, followed by one cycle at 72° C. for 10 minutes. 12-20 wells of PCR were performed for each library, in order to sample hundreds of thousands to millions of rearranged TCRβ CDR3 loci.
Example 4
Pre-Processing of Sequence Data
[0102] Sequencer data processing involves a series of steps to remove errors in the primary sequence of each read, and to compress the data. First, a complexity filter removes approximately 20% of the sequences which are misreads from the sequencer. Then, sequences were required to have a minimum of a six base match to both one of the thirteen J-regions and one of 54 V-regions. Applying the filter to the control lane containing phage sequence, on average only one sequence in 7-8 million passed these steps without false positives. Finally, a nearest neighbor algorithm was used to collapse the data into unique sequences by merging closely related sequences, in order to remove both PCR error and sequencing error (see Table 10).
Example 5
Estimating Relative CDR3 Sequence Abundance in PCR Pools and Blood samples
[0103] After collapsing the data, the underlying distribution of T-cell sequences in the blood reconstructing were derived from the sequence data. The procedure used three steps; 1) flow sorting T-cells drawn from peripheral blood, 2) PCR amplification, and 3) sequencing. Analyzing the data, the ratio of sequences in the PCR product must be derived working backward from the sequence data before estimating the true distribution of clonotypes in the blood.
[0104] For each sequence observed a given number of times in the data herein, the probability that that sequence was sampled from a particular size PCR pool is estimated. Because the CDR3 regions sequenced are sampled randomly from a massive pool of PCR products, the number of observations for each sequence are drawn from Poisson distributions. The Poisson parameters are quantized according to the number of T cell genomes that provided the template for PCR. A simple Poisson mixture model both estimates these parameters and places a pairwise probability for each sequence being drawn from each distribution. This is an expectation maximization method which reconstructs the abundances of each sequence that was drawn from the blood.
Example 6
Unseen Species Model for Estimation of True Diversity
[0105] A mixture model can reconstruct the frequency of each TCRβ CDR3 species drawn from the blood, but the larger question is how many unique CDR3 species were present in the donor? This is a fundamental question that needs to be answered as the available sample is limited in each donor, and will be more important in the future as these techniques are extrapolated to the smaller volumes of blood that can reasonably be drawn from patients undergoing treatment.
[0106] The mathematical solution provides that for a total number of TCRβ "species" or clonotypes, S, a sequencing experiment observes xs copies of sequence s. For all of the unobserved clonotypes, xs equals 0, and each TCR clonotype is "captured" in a blood draw according to a Poisson process with parameter λs. The number of T cell genomes sequenced in the first measurement 1, and in the second measurement. Since there are a large number of unique sequences, an integral will represent the sum. If G(λ) is the empirical distribution function of the parameters λ1, . . . , λs, and nx is the number of clonotypes sequenced exactly x times, then
E ( n x ) = S ∫ 0 ∞ ( - λ λ x x ! ) G ( λ ) . ##EQU00005##
[0107] The value Δ(t) is the number of new clonotypes observed in the second sequencing experiment.
Δ ( t ) = x E ( n x ) ex p 1 + ex p 2 - x E ( n x ) ex p 1 = S ∫ 0 ∞ - λ ( 1 - - λ t ) G ( λ ) ##EQU00006##
[0108] Taylor expansion of 1-e.sup.-λt gives Δ(t)=E(x1)t-E(x2)t2+E(x3)t3- . . . , which can be approximated by replacing the expectations (E(nx)) with the observed numbers in the first measurement. Using in the numbers observed in the first measurement, this formula predicts that 1.6*105 new unique sequences should be observed in the second measurement. The actual value of the second measurement was 1.8*105 new TCRβ sequences, which implies that the prediction provided a valid lower bound on total diversity. An Euler's transformation was used to regularize Δ(t) to produce a lower bound for Δ(∞).
Example 7
Error Correction and Bias Assessment
[0109] Sequence error in the primary sequence data derives primarily from two sources: (1) nucleotide misincorporation that occurs during the amplification by PCR of TCRβ CDR3 template sequences, and (2) errors in base calls introduced during sequencing of the PCR-amplified library of CDR3 sequences. The large quantity of data allows us to implement a straightforward error correcting code to correct most of the errors in the primary sequence data that are attributable to these two sources. After error correction, the number of unique, in-frame CDR3 sequences and the number of observations of each unique sequence were tabulated for each of the four flow-sorted T cell populations from the two donors. The relative frequency distribution of CDR3 sequences in the four flow cytometrically-defined populations demonstrated that antigen-experienced CD45RO+ populations contained significantly more unique CDR3 sequences with high relative frequency than the CD45RO.sup.- populations. Frequency histograms of TCRβ CDR3 sequences observed in four different T cell subsets distinguished by expression of CD4, CD8, and CD45RO and present in blood showed that ten unique sequences were each observed 200 times in the CD4+CD45RO+ (antigen-experienced) T cell sample, which was more than twice as frequent as that observed in the CD4+CD45RO.sup.- populations.
[0110] The use of a PCR step to amplify the TCRβ CDR3 regions prior to sequencing could potentially introduce a systematic bias in the inferred relative abundance of the sequences, due to differences in the efficiency of PCR amplification of CDR3 regions utilizing different Vβ and Jβ gene segments. To estimate the magnitude of any such bias, the TCRβ CDR3 regions from a sample of approximately 30,000 unique CD4+CD45RO+ T lymphocyte genomes were amplified through 25 cycles of PCR, at which point the PCR product was split in half. Half was set aside, and the other half of the PCR product was amplified for an additional 15 cycles of PCR, for a total of 40 cycles of amplification. The PCR products amplified through 25 and 40 cycles were then sequenced and compared. Over 95% of the 25 cycle sequences were also found in the 40-cycle sample: a linear correlation is observed when comparing the frequency of sequences between these samples. For sequences observed a given number of times in the 25 cycle lane, a combination of PCR bias and sampling variance accounts for the variance around the mean of the number of observations at 40 cycles. Conservatively attributing the mean variation about the line (1.5-fold) entirely to PCR bias, each cycle of PCR amplification potentially introduces a bias of average magnitude 1.51/15=1.027. Thus, the 25 cycles of PCR introduces a total bias of average magnitude 1.02725=1.95 in the inferred relative abundance of distinct CDR3 region sequences.
Example 8
JP Gene Segment Usage
[0111] The CDR3 region in each TCR 13 chain includes sequence derived from one of the thirteen J.sub.β gene segments. Analysis of the CDR3 sequences in the four different T cell populations from the two donors demonstrated that the fraction of total sequences which incorporated sequences derived from the thirteen different J.sub.β gene segments varied more than 20-fold. Jβ utilization among four different T flow cytometrically-defined T cells from a single donor is was relatively constant within a given donor. Moreover, the J.sub.β usage patterns observed in two donors, which were inferred from analysis of genomic DNA from T cells sequenced using the GA, are qualitatively similar to those observed in T cells from umbilical cord blood and from healthy adult donors, both of which were inferred from analysis of cDNA from T cells sequenced using exhaustive capillary-based techniques.
Example 9
Nucleotide Insertion Bias
[0112] Much of the diversity at the CDR3 junctions in TCR α and β chains is created by non-templated nucleotide insertions by the enzyme Terminal Deoxynucloetidyl Transferase (TdT). However, in vivo, selection plays a significant role in shaping the TCR repertoire giving rise to unpredictability. The TdT nucleotide insertion frequencies, independent of selection, were calculated using out of frame TCR sequences. These sequences are non-functional rearrangements that are carried on one allele in T cells where the second allele has a functional rearrangement. The mono-nucleotide insertion bias of TdT favors C and G (Table 11).
TABLE-US-00011 TABLE 11 Mono-nucleotide bias in out of frame data A C G T Lane 1 0.24 0.294 0.247 0.216 Lane 2 0.247 0.284 0.256 0.211 Lane 3 0.25 0.27 0.268 0.209 Lane 4 0.255 0.293 0.24 0.21
[0113] Similar nucleotide frequencies are observed in the in frame sequences (Table 12).
TABLE-US-00012 TABLE 12 Mono-nucleotide bias in in-frame data A C G T Lane 1 0.21 0.285 0.275 0.228 Lane 2 0.216 0.281 0.266 0.235 Lane 3 0.222 0.266 0.288 0.221 Lane 4 0.206 0.294 0.228 0.27
[0114] The N regions from the out of frame TCR sequences were used to measure the di-nucleotide bias. To isolate the marginal contribution of a di-nucleotide bias, the di-nucleotide frequencies was divided by the mononucleotide frequencies of each of the two bases. The measure is
m = f ( n 1 n 2 ) f ( n 1 ) f ( n 2 ) . ##EQU00007##
[0115] The matrix for m is found in Table 13.
TABLE-US-00013 TABLE 13 Di-nucleotide odd ratios for out of frame data A C G T A 1.198 0.938 0.945 0.919 C 0.988 1.172 0.88 0.931 G 0.993 0.701 1.352 0.964 T 0.784 1.232 0.767 1.23
[0116] Many of the dinucleotides are under or over represented. As an example, the odds of finding a GG pair are very high. Since the codons GGN translate to glycine, many glycines are expected in the CDR3 regions.
Example 10
Amino Acid Distributions in the CDR3 Regions
[0117] The distribution of amino acids in the CDR3 regions of TCRβ chains are shaped by the germline sequences for V, D, and J regions, the insertion bias of TdT, and selection. The distribution of amino acids in this region for the four different T cell sub-compartments is very similar between different cell subtypes. Separating the sequences into β chains of fixed length, a position dependent distribution among amino acids, which are grouped by the six chemical properties: small, special, and large hydrophobic, neutral polar, acidic and basic. The distributions are virtually identical except for the CD8+ antigen experienced T cells, which have a higher proportion of acidic bases, particularly at position 5.
[0118] Of particular interest is the comparison between CD8+ and CD4+ TCR sequences as they bind to peptides presented by class I and class II HLA molecules, respectively. The CD8+ antigen experienced T cells have a few positions with a higher proportion of acidic amino acids. This could be do binding with a basic residue found on HLA Class I molecules, but not on Class II.
Example 11
TCR β Chains with Identical Amino Acid Sequences Found in Different People
[0119] The TCR β chain sequences were translated to amino acids and then compared pairwise between the two donors. Many thousands of exact sequence matches were observed. For example, comparing the CD4+CD45RO.sup.- sub-compartments, approximately 8,000 of the 250,000 unique amino acid sequences from donor 1 were exact matches to donor 2. Many of these matching sequences at the amino acid level have multiple nucleotide differences at third codon positions. Following the example mentioned above, 1,500/8,000 identical amino acid matches had >5 nucleotide mismatches. Between any two T cell sub-types, 4-5% of the unique TCRβ sequences were found to have identical amino acid matches.
[0120] Two possibilities were examined: that 1) selection during TCR development is producing these common sequences and 2) the large bias in nucleotide insertion frequency by TdT creates similar nucleotide sequences. The in-frame pairwise matches were compared to the out-of-frame pairwise matches (see Examples 1-4, above). Changing frames preserved all of the features of the genetic code and so the same number of matches should be found if the sequence bias was responsible for the entire observation. However, almost twice as many in-frame matches as out-of-frame matches were found, suggesting that selection at the protein level is playing a significant role.
[0121] To confirm this finding of thousands of identical TCR β chain amino acid sequences, two donors were compared with respect to the CD8+CD62L+CD45RA+ (naive-like) TCRs from a third donor, a 44 year old CMV+ Caucasian female. Identical pairwise matches of many thousands of sequences at the amino acid level between the third donor and each of the original two donors were found. In contrast, 460 sequences were shared between all three donors. The large variation in total number of unique sequences between the donors is a product of the starting material and variations in loading onto the sequencer, and is not representative of a variation in true diversity in the blood of the donors.
Example 12
Higher Frequency Clonotypes are Closer to Germline
[0122] The variation in copy number between different sequences within every T cell sub-compartment ranged by a factor of over 10,000-fold. The only property that correlated with copy number was (the number of insertions plus the number of deletions), which inversely correlated. Results of the analysis showed that deletions play a smaller role than insertions in the inverse correlation with copy number.
[0123] Sequences with less insertions and deletions have receptor sequences closer to germ line. One possibility for the increased number of sequences closer to germ line is that they are the created multiple times during T cell development. Since germ line sequences are shared between people, shared TCRβ chains are likely created by TCRs with a small number of insertions and deletions.
Example 13
"Spectratype" Analysis of TCRβ CDR3 Sequences by V Gene Segment Utilization and CDR3 Length
[0124] TCR diversity has commonly been assessed using the technique of TCR spectratyping, an RT-PCR-based technique that does not assess TCR CDR3 diversity at the sequence level, but rather evaluates the diversity of TCRα or TCRβ CDR3 lengths expressed as mRNA in subsets of αβ T cells that use the same V.sub.α or V.sub.β gene segment. The spectratypes of polyclonal T cell populations with diverse repertoires of TCR CDR3 sequences, such as are seen in umbilical cord blood or in peripheral blood of healthy young adults typically contain CDR3 sequences of 8-10 different lengths that are multiples of three nucleotides, reflecting the selection for in-frame transcripts. Spectratyping also provides roughly quantitative information about the relative frequency of CDR3 sequences with each specific length. To assess whether direct sequencing of TCRβ CDR3 regions from T cell genomic DNA using the sequencer could faithfully capture all of the CDR3 length diversity that is identified by spectratyping, "virtual" TCRβ spectratypes (see Examples above) were generated from the sequence data and compared with TCRβ spectratypes generated using conventional PCR techniques. The virtual spectratypes contained all of the CDR3 length and relative frequency information present in the conventional spectratypes. Direct TCRβ CDR3 sequencing captures all of the TCR diversity information present in a conventional spectratype. A comparison of standard TCRβ spectratype data and calculated TCRβ CDR3 length distributions for sequences utilizing representative TCR Vβ gene segments and present in CD4+CD45RO+ cells from donor 1. Reducing the information contained in the sequence data to a frequency histogram of the unique CDR3 sequences with different lengths within each Vβ family readily reproduces all of the information contained in the spectratype data. In addition, the virtual spectratypes revealed the presence within each V.sub.β family of rare CDR3 sequences with both very short and very long CDR3 lengths that were not detected by conventional PCR-based spectratyping.
Example 14
Estimation of Total CDR3 Sequence Diversity
[0125] After error correction, the number of unique CDR3 sequences observed in each lane of the sequencer flow cell routinely exceeded 1×105. Given that the PCR products sequenced in each lane were necessarily derived from a small fraction of the T cell genomes present in each of the two donors, the total number of unique TCRβ CDR3 sequences in the entire T cell repertoire of each individual is likely to be far higher. Estimating the number of unique sequences in the entire repertoire, therefore, requires an estimate of the number of additional unique CDR3 sequences that exist in the blood but were not observed in the sample. The estimation of total species diversity in a large, complex population using measurements of the species diversity present in a finite sample has historically been called the "unseen species problem" (see Examples above). The solution starts with determining the number of new species, or TCRβ CDR3 sequences, that are observed if the experiment is repeated, i.e., if the sequencing is repeated on an identical sample of peripheral blood T cells, e.g., an identically prepared library of TCRβ CDR3 PCR products in a different lane of the sequencer flow cell and counting the number of new CDR3 sequences. For CD8+CD45RO.sup.- cells from donor 2, the predicted and observed number of new CDR3 sequences in a second lane are within 5% (see Examples above), suggesting that this analytic solution can, in fact, be used to estimate the total number of unique TCRβ CDR3 sequences in the entire repertoire.
[0126] The resulting estimates of the total number of unique TCRβ CDR3 sequences in the four flow cytometrically-defined T cell compartments are shown in Table 14.
TABLE-US-00014 TABLE 14 TCR repertoire diversity Donor CD8 CD4 CD45RO Diversity 1 + - + 6.3 * 105 + - - 1.24 * 106 - + + 8.2 * 105 - + - 1.28 * 106 Total T cell diversity 3.97 * 106 2 + - + 4.4 * 105 + - - 9.7 * 105 - + + 8.7 * 105 - + - 1.03 * 106 Total T cell diversity 3.31 * 106
[0127] Of note, the total TCRβ diversity in these populations is between 3-4 million unique sequences in the peripheral blood. Surprisingly, the CD45RO+, or antigen-experienced, compartment constitutes approximately 1.5 million of these sequences. This is at least an order of magnitude larger than expected. This discrepancy is likely attributable to the large number of these sequences observed at low relative frequency, which could only be detected through deep sequencing. The estimated TCRβ CDR3 repertoire sizes of each compartment in the two donors are within 20% of each other.
[0128] The results herein demonstrate that the realized TCRβ receptor diversity is at least five-fold higher than previous estimates (˜4*106 distinct CDR3 sequences), and, in particular, suggest far greater TCRβ diversity among CD45RO+ antigen-experienced αβ T cells than has previously been reported (˜1.5*106 distinct CDR3 sequences). However, bioinformatic analysis of the TCR sequence data shows strong biases in the mono- and di-nucleotide content, implying that the utilized TCR sequences are sampled from a distribution much smaller than the theoretical size. With the large diversity of TCRβ chains in each person sampled from a severely constrict space of sequences, overlap of the TCR sequence pools can be expected between each person. In fact, the results showed about 5% of CD8+ naive TCRβ chains with exact amino acid matches are shared between each pair of three different individuals. As the TCRα pool has been previously measured to be substantially smaller than the theoretical TCRβ diversity, these results show that hundreds to thousands of truly public αβ TCRs can be found.
Sequence CWU
1
1
484132DNAArtificial SequenceSynthetic DNA TRBV2 1ntcaaatttc actctgaaga
tccggtccac aa 32232DNAArtificial
SequenceSynthetic DNA TRBV3-1 2ngctcactta aatcttcaca tcaattccct gg
32327DNAArtificial SequenceSynthetic DNA
TRBV4-1 3ncttaaacct tcacctacac gccctgc
27427DNAArtificial SequenceSynthetic DNA TRBV(4-2, 4-3) 4ncttattcct
tcacctacac accctgc
27527DNAArtificial SequenceSynthetic DNA TRBV5-1 5ngctctgaga tgaatgtgag
caccttg 27627DNAArtificial
SequenceSynthetic DNA TRBV5-3 6ngctctgaga tgaatgtgag tgccttg
27727DNAArtificial SequenceSynthetic DNA
TRBV(5-4, 5-5, 5-6, 5-7, 5-8) 7ngctctgagc tgaatgtgaa cgccttg
27823DNAArtificial SequenceSynthetic DNA
TRBV6-1 8ntcgctcagg ctggagtcgg ctg
23921DNAArtificial SequenceSynthetic DNA TRBV(6-2, 6-3) 9ngctggggtt
ggagtcggct g
211022DNAArtificial SequenceSynthetic DNA TRBV6-4 10nccctcacgt tggcgtctgc
tg 221121DNAArtificial
SequenceSynthetic DNA TRBV6-5 11ngctcaggct gctgtcggct g
211222DNAArtificial SequenceSynthetic DNA
TRBV6-6 12ncgctcaggc tggagttggc tg
221323DNAArtificial SequenceSynthetic DNA TRBV6-7 13ncccctcaag
ctggagtcag ctg
231422DNAArtificial SequenceSynthetic DNA TRBV6-8 14ncactcaggc tggtgtcggc
tg 221522DNAArtificial
SequenceSynthetic DNA TRBV6-9 15ncgctcaggc tggagtcagc tg
221625DNAArtificial SequenceSynthetic DNA
TRBV7-1 16nccactctga agttccagcg cacac
251724DNAArtificial SequenceSynthetic DNA TRBV7-2 17ncactctgac
gatccagcgc acac
241827DNAArtificial SequenceSynthetic DNA TRBV7-3 18nctctactct gaagatccag
cgcacag 271925DNAArtificial
SequenceSynthetic DNA TRBV7-4 19nccactctga agatccagcg cacag
252024DNAArtificial SequenceSynthetic DNA
TRBV7-6 20ncactctgac gatccagcgc acag
242125DNAArtificial SequenceSynthetic DNA TRBV7-7 21nccactctga
cgattcagcg cacag
252225DNAArtificial SequenceSynthetic DNA TRBV7-8 22nccactctga agatccagcg
cacac 252324DNAArtificial
SequenceSynthetic DNA TRBV7-9 23ncaccttgga gatccagcgc acag
242429DNAArtificial SequenceSynthetic DNA
TRBV9 24ngcactctga actaaacctg agctctctg
292523DNAArtificial SequenceSynthetic DNA TRBV10-1 25ncccctcact
ctggagtctg ctg
232624DNAArtificial SequenceSynthetic DNA TRBV10-2 26nccccctcac
tctggagtca gcta
242724DNAArtificial SequenceSynthetic DNA TRBV10-3 27ncctcctcac
tctggagtcc gcta
242825DNAArtificial SequenceSynthetic DNA TRBV(11-1, 11-3) 28nccactctca
agatccagcc tgcag
252927DNAArtificial SequenceSynthetic DNA TRBV11-2 29nctccactct
caagatccag cctgcaa
273025DNAArtificial SequenceSynthetic DNA TRBV(12-3, 12-4, 12-5)
30nccactctga agatccagcc ctcag
253129DNAArtificial SequenceSynthetic DNA TRBV13 31ncattctgaa ctgaacatga
gctccttgg 293225DNAArtificial
SequenceSynthetic DNA TRBV14 32nctactctga aggtgcagcc tgcag
253329DNAArtificial SequenceSynthetic DNA
TRBV15 33ngataacttc caatccagga ggccgaaca
293427DNAArtificial SequenceSynthetic DNA TRBV16 34nctgtagcct
tgagatccag gctacga
273525DNAArtificial SequenceSynthetic DNA TRBV17 35ncttccacgc tgaagatcca
tcccg 253625DNAArtificial
SequenceSynthetic DNA TRBV18 36ngcatcctga ggatccagca ggtag
253723DNAArtificial SequenceSynthetic DNA
TRBV19 37ncctctcact gtgacatcgg ccc
233827DNAArtificial SequenceSynthetic DNA TRBV20-1 38ncttgtccac
tctgacagtg accagtg
273924DNAArtificial SequenceSynthetic DNA TRBV23-1 39ncagcctggc
aatcctgtcc tcag
244025DNAArtificial SequenceSynthetic DNA TRBV24-1 40nctccctgtc
cctagagtct gccat
254122DNAArtificial SequenceSynthetic DNA TRBV25-1 41nccctgaccc
tggagtctgc ca
224222DNAArtificial SequenceSynthetic DNA TRBV27 42nccctgatcc tggagtcgcc
ca 224324DNAArtificial
SequenceSynthetic DNA TRBV28 43nctccctgat tctggagtcc gcca
244432DNAArtificial SequenceSynthetic DNA
TRBV29-1 44nctaacattc tcaactctga ctgtgagcaa ca
324529DNAArtificial SequenceSynthetic DNA TRBV30 45ncggcagttc
atcctgagtt ctaagaagc
294636DNAArtificial SequenceSynthetic DNA TRBJ1-1 46nttacctaca actgtgagtc
tggtgccttg tccaaa 364734DNAArtificial
SequenceSynthetic DNA TRBJ1-2 47nacctacaac ggttaacctg gtccccgaac cgaa
344834DNAArtificial SequenceSynthetic DNA
TRBJ1-3 48nacctacaac agtgagccaa cttccctctc caaa
344932DNAArtificial SequenceSynthetic DNA TRBJ1-4 49nccaagacag
agagctgggt tccactgcca aa
325032DNAArtificial SequenceSynthetic DNA TRBJ1-6 50nctgtcacag tgagcctggt
cccgttccca aa 325126DNAArtificial
SequenceSynthetic DNA TRBJ2-1 51ncggtgagcc gtgtccctgg cccgaa
265232DNAArtificial SequenceSynthetic DNA
TRBJ2-2 52nccagtacgg tcagcctaga gccttctcca aa
325327DNAArtificial SequenceSynthetic DNA TRBJ2-3 53nactgtcagc
cgggtgcctg ggccaaa
275423DNAArtificial SequenceSynthetic DNA TRBJ2-4 54nagagccggg tcccggcgcc
gaa 235523DNAArtificial
SequenceSynthetic DNA TRBJ2-5 55nggagccgcg tgcctggccc gaa
235624DNAArtificial SequenceSynthetic DNA
TRBJ2-6 56ngtcagcctg ctgccggccc cgaa
245724DNAArtificial SequenceSynthetic DNA TRBJ2-7 57ngtgagcctg
gtgcccggcc cgaa
245865DNAArtificial SequenceSynthetic DNA TRBV2 58caagcagaag acggcatacg
agctcttccg atcttcaaat ttcactctga agatccggtc 60cacaa
655965DNAArtificial
SequenceSynthetic DNA TRBV3-1 59caagcagaag acggcatacg agctcttccg
atctgctcac ttaaatcttc acatcaattc 60cctgg
656060DNAArtificial SequenceSynthetic
DNA TRBV4-1 60caagcagaag acggcatacg agctcttccg atctcttaaa ccttcaccta
cacgccctgc 606160DNAArtificial SequenceSynthetic DNA TRBV(4-2, 4-3)
61caagcagaag acggcatacg agctcttccg atctcttatt ccttcaccta cacaccctgc
606260DNAArtificial SequenceSynthetic DNA TRBV5-1 62caagcagaag acggcatacg
agctcttccg atctgctctg agatgaatgt gagcaccttg 606360DNAArtificial
SequenceSynthetic DNA TRBV5-3 63caagcagaag acggcatacg agctcttccg
atctgctctg agatgaatgt gagtgccttg 606460DNAArtificial
SequenceSynthetic DNA TRBV(5-4, 5-5, 5-6, 5-7, 5-8) 64caagcagaag
acggcatacg agctcttccg atctgctctg agctgaatgt gaacgccttg
606556DNAArtificial SequenceSynthetic DNA TRBV6-1 65caagcagaag acggcatacg
agctcttccg atcttcgctc aggctggagt cggctg 566654DNAArtificial
SequenceSynthetic DNA TRBV(6-2, 6-3) 66caagcagaag acggcatacg agctcttccg
atctgctggg gttggagtcg gctg 546755DNAArtificial
SequenceSynthetic DNA TRBV6-4 67caagcagaag acggcatacg agctcttccg
atctccctca cgttggcgtc tgctg 556854DNAArtificial
SequenceSynthetic DNA TRBV6-5 68caagcagaag acggcatacg agctcttccg
atctgctcag gctgctgtcg gctg 546955DNAArtificial
SequenceSynthetic DNA TRBV6-6 69caagcagaag acggcatacg agctcttccg
atctcgctca ggctggagtt ggctg 557056DNAArtificial
SequenceSynthetic DNA TRBV6-7 70caagcagaag acggcatacg agctcttccg
atctcccctc aagctggagt cagctg 567155DNAArtificial
SequenceSynthetic DNA TRBV6-8 71caagcagaag acggcatacg agctcttccg
atctcactca ggctggtgtc ggctg 557255DNAArtificial
SequenceSynthetic DNA TRBV6-9 72caagcagaag acggcatacg agctcttccg
atctcgctca ggctggagtc agctg 557358DNAArtificial
SequenceSynthetic DNA TRBV7-1 73caagcagaag acggcatacg agctcttccg
atctccactc tgaagttcca gcgcacac 587457DNAArtificial
SequenceSynthetic DNA TRBV7-2 74caagcagaag acggcatacg agctcttccg
atctcactct gacgatccag cgcacac 577560DNAArtificial
SequenceSynthetic DNA TRBV7-3 75caagcagaag acggcatacg agctcttccg
atctctctac tctgaagatc cagcgcacag 607658DNAArtificial
SequenceSynthetic DNA TRBV7-4 76caagcagaag acggcatacg agctcttccg
atctccactc tgaagatcca gcgcacag 587757DNAArtificial
SequenceSynthetic DNA TRBV7-6 77caagcagaag acggcatacg agctcttccg
atctcactct gacgatccag cgcacag 577858DNAArtificial
SequenceSynthetic DNA TRBV7-7 78caagcagaag acggcatacg agctcttccg
atctccactc tgacgattca gcgcacag 587958DNAArtificial
SequenceSynthetic DNA TRBV7-8 79caagcagaag acggcatacg agctcttccg
atctccactc tgaagatcca gcgcacac 588057DNAArtificial
SequenceSynthetic DNA TRBV7-9 80caagcagaag acggcatacg agctcttccg
atctcacctt ggagatccag cgcacag 578162DNAArtificial
SequenceSynthetic DNA TRBV9 81caagcagaag acggcatacg agctcttccg atctgcactc
tgaactaaac ctgagctctc 60tg
628256DNAArtificial SequenceSynthetic DNA
TRBV10-1 82caagcagaag acggcatacg agctcttccg atctcccctc actctggagt ctgctg
568357DNAArtificial SequenceSynthetic DNA TRBV10-2 83caagcagaag
acggcatacg agctcttccg atctccccct cactctggag tcagcta
578457DNAArtificial SequenceSynthetic DNA TRBV10-3 84caagcagaag
acggcatacg agctcttccg atctcctcct cactctggag tccgcta
578558DNAArtificial SequenceSynthetic DNA TRBV(11-1, 11-3) 85caagcagaag
acggcatacg agctcttccg atctccactc tcaagatcca gcctgcag
588660DNAArtificial SequenceSynthetic DNA TRBV11-2 86caagcagaag
acggcatacg agctcttccg atctctccac tctcaagatc cagcctgcaa
608758DNAArtificial SequenceSynthetic DNA TRBV(12-3, 12-4, 12-5)
87caagcagaag acggcatacg agctcttccg atctccactc tgaagatcca gccctcag
588862DNAArtificial SequenceSynthetic DNA TRBV13 88caagcagaag acggcatacg
agctcttccg atctcattct gaactgaaca tgagctcctt 60gg
628958DNAArtificial
SequenceSynthetic DNA TRBV14 89caagcagaag acggcatacg agctcttccg
atctctactc tgaaggtgca gcctgcag 589062DNAArtificial
SequenceSynthetic DNA TRBV15 90caagcagaag acggcatacg agctcttccg
atctgataac ttccaatcca ggaggccgaa 60ca
629160DNAArtificial SequenceSynthetic
DNA TRBV16 91caagcagaag acggcatacg agctcttccg atctctgtag ccttgagatc
caggctacga 609258DNAArtificial SequenceSynthetic DNA TRBV17
92caagcagaag acggcatacg agctcttccg atctcttcca cgctgaagat ccatcccg
589358DNAArtificial SequenceSynthetic DNA TRBV18 93caagcagaag acggcatacg
agctcttccg atctgcatcc tgaggatcca gcaggtag 589456DNAArtificial
SequenceSynthetic DNA TRBV19 94caagcagaag acggcatacg agctcttccg
atctcctctc actgtgacat cggccc 569560DNAArtificial
SequenceSynthetic DNA TRBV20-1 95caagcagaag acggcatacg agctcttccg
atctcttgtc cactctgaca gtgaccagtg 609657DNAArtificial
SequenceSynthetic DNA TRBV23-1 96caagcagaag acggcatacg agctcttccg
atctcagcct ggcaatcctg tcctcag 579758DNAArtificial
SequenceSynthetic DNA TRBV24-1 97caagcagaag acggcatacg agctcttccg
atctctccct gtccctagag tctgccat 589855DNAArtificial
SequenceSynthetic DNA TRBV25-1 98caagcagaag acggcatacg agctcttccg
atctccctga ccctggagtc tgcca 559955DNAArtificial
SequenceSynthetic DNA TRBV27 99caagcagaag acggcatacg agctcttccg
atctccctga tcctggagtc gccca 5510057DNAArtificial
SequenceSynthetic DNA TRBV28 100caagcagaag acggcatacg agctcttccg
atctctccct gattctggag tccgcca 5710165DNAArtificial
SequenceSynthetic DNA TRBV29-1 101caagcagaag acggcatacg agctcttccg
atctctaaca ttctcaactc tgactgtgag 60caaca
6510262DNAArtificial SequenceSynthetic
DNA TRBV30 102caagcagaag acggcatacg agctcttccg atctcggcag ttcatcctga
gttctaagaa 60gc
6210360DNAArtificial SequenceSynthetic DNA TRBJ1-1
103aatgatacgg cgaccaccga gatctttacc tacaactgtg agtctggtgc cttgtccaaa
6010458DNAArtificial SequenceSynthetic DNA TRBJ1-3 104aatgatacgg
cgaccaccga gatctaccta caacagtgag ccaacttccc tctccaaa
5810556DNAArtificial SequenceSynthetic DNA TRBJ1-4 105aatgatacgg
cgaccaccga gatctccaag acagagagct gggttccact gccaaa
5610656DNAArtificial SequenceSynthetic DNA TRBJ1-6 106aatgatacgg
cgaccaccga gatctctgtc acagtgagcc tggtcccgtt cccaaa
5610750DNAArtificial SequenceSynthetic DNA TRBJ2-1 107aatgatacgg
cgaccaccga gatctcggtg agccgtgtcc ctggcccgaa
5010856DNAArtificial SequenceSynthetic DNA TRBJ2-2 108aatgatacgg
cgaccaccga gatctccagt acggtcagcc tagagccttc tccaaa
5610951DNAArtificial SequenceSynthetic DNA TRBJ2-3 109aatgatacgg
cgaccaccga gatctactgt cagccgggtg cctgggccaa a
5111047DNAArtificial SequenceSynthetic DNA TRBJ2-4 110aatgatacgg
cgaccaccga gatctagagc cgggtcccgg cgccgaa
4711147DNAArtificial SequenceSynthetic DNA TRBJ2-5 111aatgatacgg
cgaccaccga gatctggagc cgcgtgcctg gcccgaa
4711248DNAArtificial SequenceSynthetic DNA TRBJ2-6 112aatgatacgg
cgaccaccga gatctgtcag cctgctgccg gccccgaa
4811348DNAArtificial SequenceSynthetic DNA TRBJ2-7 113aatgatacgg
cgaccaccga gatctgtgag cctggtgccc ggcccgaa
48114284DNAArtificial SequenceSynthetic DNA TRBV1*01 114gatactggaa
ttacccagac accaaaatac ctggtcacag caatggggag taaaaggaca 60atgaaacgtg
agcatctggg acatgattct atgtattggt acagacagaa agctaagaaa 120tccctggagt
tcatgtttta ctacaactgt aaggaattca ttgaaaacaa gactgtgcca 180aatcacttca
cacctgaatg ccctgacagc tctcgcttat accttcatgt ggtcgcactg 240cagcaagaag
actcagctgc gtatctctgc accagcagcc aaga
284115290DNAArtificial SequenceSynthetic DNA TRBV2*01 115gaacctgaag
tcacccagac tcccagccat caggtcacac agatgggaca ggaagtgatc 60ttgcgctgtg
tccccatctc taatcactta tacttctatt ggtacagaca aatcttgggg 120cagaaagtcg
agtttctggt ttccttttat aataatgaaa tctcagagaa gtctgaaata 180ttcgatgatc
aattctcagt tgaaaggcct gatggatcaa atttcactct gaagatccgg 240tccacaaagc
tggaggactc agccatgtac ttctgtgcca gcagtgaagc
290116288DNAArtificial SequenceSynthetic DNA TRBV2*03 116gaacctgaag
tcacccagac tcccagccat caggtcacac agatgggaca ggaagtgatc 60ttgcgctgtg
tccccatctc taatcactta tacttctatt ggtacagaca aatcttgggg 120cagaaagtcg
agtttctggt ttccttttat aataatgaaa tctcagagaa gtctgaaata 180ttcgatgatc
aattctcagt tgagaggcct gatggatcaa atttcactct gaagatccgg 240tccacaaagc
tggaggactc agccatgtac ttctgtgcca gcagtgaa
288117287DNAArtificial SequenceSynthetic DNA TRBV3-1*01 117gacacagctg
tttcccagac tccaaaatac ctggtcacac agatgggaaa cgacaagtcc 60attaaatgtg
aacaaaatct gggccatgat actatgtatt ggtataaaca ggactctaag 120aaatttctga
agataatgtt tagctacaat aataaggagc tcattataaa tgaaacagtt 180ccaaatcgct
tctcacctaa atctccagac aaagctcact taaatcttca catcaattcc 240ctggagcttg
gtgactctgc tgtgtatttc tgtgccagca gccaaga
287118279DNAArtificial SequenceSynthetic DNA TRBV3-1*02 118gacacagctg
tttcccagac tccaaaatac ctggtcacac agatgggaaa cgacaagtcc 60attaaatgtg
aacaaaatct gggccatgat actatgtatt ggtataaaca ggactctaag 120aaatttctga
agataatgtt tagctacaat aacaaggaga tcattataaa tgaaacagtt 180ccaaatcgat
tctcacctaa atctccagac aaagctaaat taaatcttca catcaattcc 240ctggagcttg
gtgactctgc tgtgtatttc tgtgccagc
279119287DNAArtificial SequenceSynthetic DNA TRBV3-2*01 119gacacagccg
tttcccagac tccaaaatac ctggtcacac agatgggaaa aaaggagtct 60cttaaatgag
aacaaaatct gggccataat gctatgtatt ggtataaaca ggactctaag 120aaatttctga
agacaatgtt tatctacagt aacaaggagc caattttaaa tgaaacagtt 180ccaaatcgct
tctcacctga ctctccagac aaagctcatt taaatcttca catcaattcc 240ctggagcttg
gtgactctgc tgtgtatttc tgtgccagca gccaaga
287120287DNAArtificial SequenceSynthetic DNA TRBV3-2*02 120gacacagccg
tttcccagac tccaaaatac ctggtcacac agatgggaaa aaaggagtct 60cttaaatgag
aacaaaatct gggccataat gctatgtatt ggtataaaca ggactctaag 120aaatttctga
agacaatgtt tatctacagt aacaaggagc caattttaaa tgaaacagtt 180ccaaatcgct
tctcacctga ctctccagac aaagttcatt taaatcttca catcaattcc 240ctggagcttg
gtgactctgc tgtgtatttc tgtgccagca gccaaga
287121285DNAArtificial SequenceSynthetic DNA TRBV3-2*03 121gacacagccg
tttcccagac tccaaaatac ctggtcacac agacgggaaa aaaggagtct 60cttaaatgag
aacaaaatct gggccataat gctatgtatt ggtataaaca ggactctaag 120aaatttctga
agacaatgtt tatctacagt aacaaggagc caattttaaa tgaaacagtt 180ccaaatcgct
tctcacctga ctctccagac aaagttcatt taaatcttca catcaattcc 240ctggagcttg
gtgactctgc tgtgtatttc tgtgccagca gccaa
285122287DNAArtificial SequenceSynthetic DNA TRBV4-1*01 122gacactgaag
ttacccagac accaaaacac ctggtcatgg gaatgacaaa taagaagtct 60ttgaaatgtg
aacaacatat ggggcacagg gctatgtatt ggtacaagca gaaagctaag 120aagccaccgg
agctcatgtt tgtctacagc tatgagaaac tctctataaa tgaaagtgtg 180ccaagtcgct
tctcacctga atgccccaac agctctctct taaaccttca cctacacgcc 240ctgcagccag
aagactcagc cctgtatctc tgcgccagca gccaaga
287123258DNAArtificial SequenceSynthetic DNA TRBV4-1*02 123cacctggtca
tgggaatgac aaataagaag tctttgaaat gtgaacaaca tatggggcac 60agggcaatgt
attggtacaa gcagaaagct aagaagccac cggagctcat gtttgtctac 120agctatgaga
aactctctat aaatgaaagt gtgccaagtc gcttctcacc tgaatgcccc 180aacagctctc
tcttaaacct tcacctacac gccctgcagc cagaagactc agccctgtat 240ctctgcgcca
gcagccaa
258124287DNAArtificial SequenceSynthetic DNA TRBV4-2*01 124gaaacgggag
ttacgcagac accaagacac ctggtcatgg gaatgacaaa taagaagtct 60ttgaaatgtg
aacaacatct ggggcataac gctatgtatt ggtacaagca aagtgctaag 120aagccactgg
agctcatgtt tgtctacaac tttaaagaac agactgaaaa caacagtgtg 180ccaagtcgct
tctcacctga atgccccaac agctctcact tattccttca cctacacacc 240ctgcagccag
aagactcggc cctgtatctc tgtgccagca gccaaga
287125282DNAArtificial SequenceSynthetic DNA TRBV4-2*02 125gaaacgggag
ttacgcagac accaagacac ctggtcatgg gaatgacaaa taagaagtct 60ttgaaatgtg
aacaacatct ggggcataac gctatgtatt ggtacaagca aagtgctaag 120aagccactgg
agctcatgtt tgtctacaac tttaaagaac agactgaaaa caacagtgtg 180ccaagtcgct
tctcacctga atgccccaac agctctcact tatgccttca cctacacacc 240ctgcagccag
aagactcggc cctgtatctc tgtgccagca cc
282126287DNAArtificial SequenceSynthetic DNA TRBV4-3*01 126gaaacgggag
ttacgcagac accaagacac ctggtcatgg gaatgacaaa taagaagtct 60ttgaaatgtg
aacaacatct gggtcataac gctatgtatt ggtacaagca aagtgctaag 120aagccactgg
agctcatgtt tgtctacagt cttgaagaac gggttgaaaa caacagtgtg 180ccaagtcgct
tctcacctga atgccccaac agctctcact tattccttca cctacacacc 240ctgcagccag
aagactcggc cctgtatctc tgcgccagca gccaaga
287127282DNAArtificial SequenceSynthetic DNA TRBV4-3*02 127gaaacgggag
ttacgcagac accaagacac ctggtcatgg gaatgacaaa taagaagtct 60ttgaaatgtg
aacaacatct gggtcataac gctatgtatt ggtacaagca aagtgctaag 120aagccactgg
agctcatgtt tgtctacagt cttgaagaac gggttgaaaa caacagtgtg 180ccaagtcgct
tctcacctga atgccccaac agctctcact tatcccttca cctacacacc 240ctgcagccag
aagactcggc cctgtatctc tgcgccagca gc
282128282DNAArtificial SequenceSynthetic DNA TRBV4-3*03 128gaaacgggag
ttacgcagac accaagacac ctggtcatgg gaatgacaaa taagaagtct 60ttgaaatgtg
aacaacatct gggtcataac gctatgtatt ggtacaagca aagtgctaag 120aagccactgg
agctcatgtt tgtctacagt cttgaagaac gtgttgaaaa caacagtgtg 180ccaagtcgct
tctcacctga atgccccaac agctctcact tattccttca cctacacacc 240ctgcagccag
aagactcggc cctgtatctc tgcgccagca gc
282129231DNAArtificial SequenceSynthetic DNA TRBV4-3*04 129aagaagtctt
tgaaatgtga acaacatctg gggcataacg ctatgtattg gtacaagcaa 60agtgctaaga
agccactgga gctcatgttt gtctacagtc ttgaagaacg ggttgaaaac 120aacagtgtgc
caagtcgctt ctcacctgaa tgccccaaca gctctcactt attccttcac 180ctacacaccc
tgcagccaga agactcggcc ctgtatctct gcgccagcag c
231130286DNAArtificial SequenceSynthetic DNA TRBV5-1*01 130aaggctggag
tcactcaaac tccaagatat ctgatcaaaa cgagaggaca gcaagtgaca 60ctgagctgct
cccctatctc tgggcatagg agtgtatcct ggtaccaaca gaccccagga 120cagggccttc
agttcctctt tgaatacttc agtgagacac agagaaacaa aggaaacttc 180cctggtcgat
tctcagggcg ccagttctct aactctcgct ctgagatgaa tgtgagcacc 240ttggagctgg
gggactcggc cctttatctt tgcgccagca gcttgg
286131285DNAArtificial SequenceSynthetic DNA TRBV5-1*02 131agggctgggg
tcactcaaac tccaagacat ctgatcaaaa cgagaggaca gcaagtgaca 60ctgggctgct
cccctatctc tgggcatagg agtgtatcct ggtaccaaca gaccctagga 120cagggccttc
agttcctctt tgaatacttc agtgagacac agagaaacaa aggaaacttc 180cttggtcgat
tctcagggcg ccagttctct aactctcgct ctgagatgaa tgtgagcacc 240ttggagctgg
gggactcggc cctttatctt tgcgccagcg cttgc
285132286DNAArtificial SequenceSynthetic DNA TRBV5-3*01 132gaggctggag
tcacccaaag tcccacacac ctgatcaaaa cgagaggaca gcaagtgact 60ctgagatgct
ctcctatctc tgggcacagc agtgtgtcct ggtaccaaca ggccccgggt 120caggggcccc
agtttatctt tgaatatgct aatgagttaa ggagatcaga aggaaacttc 180cctaatcgat
tctcagggcg ccagttccat gactgttgct ctgagatgaa tgtgagtgcc 240ttggagctgg
gggactcggc cctgtatctc tgtgccagaa gcttgg
286133286DNAArtificial SequenceSynthetic DNA TRBV5-3*02 133gaggctggag
tcacccaaag tcccacacac ctgatcaaaa cgagaggaca gcaagtgact 60ctgagatgct
ctcctatctc tgggcacagc agtgtgtcct ggtaccaaca ggccccgggt 120caggggcccc
agtttatctt tgaatatgct aatgagttaa ggagatcaga aggaaacttc 180cctaatcgat
tctcagggcg ccagttccat gactattgct ctgagatgaa tgtgagtgcc 240ttggagctgg
gggactcggc cctgtatctc tgtgccagaa gcttgg
286134286DNAArtificial SequenceSynthetic DNA TRBV5-4*01 134gagactggag
tcacccaaag tcccacacac ctgatcaaaa cgagaggaca gcaagtgact 60ctgagatgct
cttctcagtc tgggcacaac actgtgtcct ggtaccaaca ggccctgggt 120caggggcccc
agtttatctt tcagtattat agggaggaag agaatggcag aggaaacttc 180cctcctagat
tctcaggtct ccagttccct aattatagct ctgagctgaa tgtgaacgcc 240ttggagctgg
acgactcggc cctgtatctc tgtgccagca gcttgg
286135282DNAArtificial SequenceSynthetic DNA TRBV5-4*02 135gagactggag
tcacccaaag tcccacacac ctgatcaaaa cgagaggaca gcaagtgact 60ctgagatgct
cttctcagtc tgggcacaac actgtgtcct ggtaccaaca ggccctgggt 120caggggcccc
agtttatctt tcagtattat agggaggaag agaatggcag aggaaacttc 180cctcctagat
tctcaggtct ccagttccct aattataact ctgagctgaa tgtgaacgcc 240ttggagctgg
acgactcggc cctgtatctc tgtgccagca gc
282136234DNAArtificial SequenceSynthetic DNA TRBV5-4*03 136cagcaagtga
cactgagatg ctcttctcag tctgggcaca acactgtgtc ctggtaccaa 60caggccctgg
gtcaggggcc ccagtttatc tttcagtatt atagggagga agagaatggc 120agaggaaact
tccctcctag attctcaggt ctccagttcc ctaattatag ctctgagctg 180aatgtgaacg
ccttggagct ggacgactcg gccctgtatc tctgtgccag cagc
234137192DNAArtificial SequenceSynthetic DNA TRBV5-4*04 137actgtgtcct
ggtaccaaca ggccctgggt caggggcccc agtttatctt tcagtattat 60agggaggaag
agaatggcag aggaaactcc cctcctagat tctcaggtct ccagttccct 120aattatagct
ctgagctgaa tgtgaacgcc ttggagctgg acgactcggc cctgtatctc 180tgtgccagca
gc
192138286DNAArtificial SequenceSynthetic DNA TRBV5-5*01 138gacgctggag
tcacccaaag tcccacacac ctgatcaaaa cgagaggaca gcaagtgact 60ctgagatgct
ctcctatctc tgggcacaag agtgtgtcct ggtaccaaca ggtcctgggt 120caggggcccc
agtttatctt tcagtattat gagaaagaag agagaggaag aggaaacttc 180cctgatcgat
tctcagctcg ccagttccct aactatagct ctgagctgaa tgtgaacgcc 240ttgttgctgg
gggactcggc cctgtatctc tgtgccagca gcttgg
286139282DNAArtificial SequenceSynthetic DNA TRBV5-5*02 139gacgctggag
tcacccaaag tcccacacac ctgatcaaaa cgagaggaca gcacgtgact 60ctgagatgct
ctcctatctc tgggcacaag agtgtgtcct ggtaccaaca ggtcctgggt 120caggggcccc
agtttatctt tcagtattat gagaaagaag agagaggaag aggaaacttc 180cctgatcgat
tctcagctcg ccagttccct aactatagct ctgagctgaa tgtgaacgcc 240ttgttgctgg
gggactcggc cctgtatctc tgtgccagca gc
282140282DNAArtificial SequenceSynthetic DNA TRBV5-5*03 140gacgctggag
tcacccaaag tcccacacac ctgatcaaaa cgagaggaca gcaagtgact 60ctgagatgct
ctcctatctc tgagcacaag agtgtgtcct ggtaccaaca ggtcctgggt 120caggggcccc
agtttatctt tcagtattat gagaaagaag agagaggaag aggaaacttc 180cctgatcgat
tctcagctcg ccagttccct aactatagct ctgagctgaa tgtgaacgcc 240ttgttgctgg
gggactcggc cctgtatctc tgtgccagca gc
282141286DNAArtificial SequenceSynthetic DNA TRBV5-6*01 141gacgctggag
tcacccaaag tcccacacac ctgatcaaaa cgagaggaca gcaagtgact 60ctgagatgct
ctcctaagtc tgggcatgac actgtgtcct ggtaccaaca ggccctgggt 120caggggcccc
agtttatctt tcagtattat gaggaggaag agagacagag aggcaacttc 180cctgatcgat
tctcaggtca ccagttccct aactatagct ctgagctgaa tgtgaacgcc 240ttgttgctgg
gggactcggc cctctatctc tgtgccagca gcttgg
286142286DNAArtificial SequenceSynthetic DNA TRBV5-7*01 142gacgctggag
tcacccaaag tcccacacac ctgatcaaaa cgagaggaca gcacgtgact 60ctgagatgct
ctcctatctc tgggcacacc agtgtgtcct cgtaccaaca ggccctgggt 120caggggcccc
agtttatctt tcagtattat gagaaagaag agagaggaag aggaaacttc 180cctgatcaat
tctcaggtca ccagttccct aactatagct ctgagctgaa tgtgaacgcc 240ttgttgctag
gggactcggc cctctatctc tgtgccagca gcttgg
286143286DNAArtificial SequenceSynthetic DNA TRBV5-8*01 143gaggctggag
tcacacaaag tcccacacac ctgatcaaaa cgagaggaca gcaagcgact 60ctgagatgct
ctcctatctc tgggcacacc agtgtgtact ggtaccaaca ggccctgggt 120ctgggcctcc
agttcctcct ttggtatgac gagggtgaag agagaaacag aggaaacttc 180cctcctagat
tttcaggtcg ccagttccct aattatagct ctgagctgaa tgtgaacgcc 240ttggagctgg
aggactcggc cctgtatctc tgtgccagca gcttgg
286144238DNAArtificial SequenceSynthetic DNA TRBV5-8*02 144aggacagcaa
gcgactctga gatgctctcc tatctctggg cacaccagtg tgtactggta 60ccaacaggcc
ctgggtctgg gcctccagct cctcctttgg tatgacgagg gtgaagagag 120aaacagagga
aacttccctc ctagattttc aggtcgccag ttccctaatt atagctctga 180gctgaatgtg
aacgccttgg agctggagga ctcggccctg tatctctgtg ccagcagc
238145287DNAArtificial SequenceSynthetic DNA TRBV6-1*01 145aatgctggtg
tcactcagac cccaaaattc caggtcctga agacaggaca gagcatgaca 60ctgcagtgtg
cccaggatat gaaccataac tccatgtact ggtatcgaca agacccaggc 120atgggactga
ggctgattta ttactcagct tctgagggta ccactgacaa aggagaagtc 180cccaatggct
acaatgtctc cagattaaac aaacgggagt tctcgctcag gctggagtcg 240gctgctccct
cccagacatc tgtgtacttc tgtgccagca gtgaagc
287146287DNAArtificial SequenceSynthetic DNA TRBV6-2*01 146aatgctggtg
tcactcagac cccaaaattc cgggtcctga agacaggaca gagcatgaca 60ctgctgtgtg
cccaggatat gaaccatgaa tacatgtact ggtatcgaca agacccaggc 120atggggctga
ggctgattca ttactcagtt ggtgagggta caactgccaa aggagaggtc 180cctgatggct
acaatgtctc cagattaaaa aaacagaatt tcctgctggg gttggagtcg 240gctgctccct
cccaaacatc tgtgtacttc tgtgccagca gttactc
287147287DNAArtificial SequenceSynthetic DNA TRBV6-3*01 147aatgctggtg
tcactcagac cccaaaattc cgggtcctga agacaggaca gagcatgaca 60ctgctgtgtg
cccaggatat gaaccatgaa tacatgtact ggtatcgaca agacccaggc 120atggggctga
ggctgattca ttactcagtt ggtgagggta caactgccaa aggagaggtc 180cctgatggct
acaatgtctc cagattaaaa aaacagaatt tcctgctggg gttggagtcg 240gctgctccct
cccaaacatc tgtgtacttc tgtgccagca gttactc
287148287DNAArtificial SequenceSynthetic DNA TRBV6-4*01 148attgctggga
tcacccaggc accaacatct cagatcctgg cagcaggacg gcgcatgaca 60ctgagatgta
cccaggatat gagacataat gccatgtact ggtatagaca agatctagga 120ctggggctaa
ggctcatcca ttattcaaat actgcaggta ccactggcaa aggagaagtc 180cctgatggtt
atagtgtctc cagagcaaac acagatgatt tccccctcac gttggcgtct 240gctgtaccct
ctcagacatc tgtgtacttc tgtgccagca gtgactc
287149287DNAArtificial SequenceSynthetic DNA TRBV6-4*02 149actgctggga
tcacccaggc accaacatct cagatcctgg cagcaggacg gagcatgaca 60ctgagatgta
cccaggatat gagacataat gccatgtact ggtatagaca agatctagga 120ctggggctaa
ggctcatcca ttattcaaat actgcaggta ccactggcaa aggagaagtc 180cctgatggtt
atagtgtctc cagagcaaac acagatgatt tccccctcac gttggcgtct 240gctgtaccct
ctcagacatc tgtgtacttc tgtgccagca gtgactc
287150287DNAArtificial SequenceSynthetic DNA TRBV6-5*01 150aatgctggtg
tcactcagac cccaaaattc caggtcctga agacaggaca gagcatgaca 60ctgcagtgtg
cccaggatat gaaccatgaa tacatgtcct ggtatcgaca agacccaggc 120atggggctga
ggctgattca ttactcagtt ggtgctggta tcactgacca aggagaagtc 180cccaatggct
acaatgtctc cagatcaacc acagaggatt tcccgctcag gctgctgtcg 240gctgctccct
cccagacatc tgtgtacttc tgtgccagca gttactc
287151287DNAArtificial SequenceSynthetic DNA TRBV6-6*01 151aatgctggtg
tcactcagac cccaaaattc cgcatcctga agataggaca gagcatgaca 60ctgcagtgta
cccaggatat gaaccataac tacatgtact ggtatcgaca agacccaggc 120atggggctga
agctgattta ttattcagtt ggtgctggta tcactgataa aggagaagtc 180ccgaatggct
acaacgtctc cagatcaacc acagaggatt tcccgctcag gctggagttg 240gctgctccct
cccagacatc tgtgtacttc tgtgccagca gttactc
287152282DNAArtificial SequenceSynthetic DNA TRBV6-6*02 152aatgctggtg
tcactcagac cccaaaattc cgcatcctga agataggaca gagcatgaca 60ctgcagtgtg
cccaggatat gaaccataac tacatgtact ggtatcgaca agacccaggc 120atggggctga
agctgattta ttattcagtt ggtgctggta tcactgacaa aggagaagtc 180ccgaatggct
acaacgtctc cagatcaacc acagaggatt tcccgctcag gctggagttg 240gctgctccct
cccagacatc tgtgtacttc tgtgccagca gt
282153282DNAArtificial SequenceSynthetic DNA TRBV6-6*03 153aatgctggtg
tcactcagac cccaaaattc cgcatcctga agataggaca gagcatgaca 60ctgcagtgtg
cccaggatat gaaccataac tacatgtact ggtatcgaca agacccaggc 120atggggctga
agctgattta ttattcagtt ggtgctggta tcactgataa aggagaagtc 180ccgaatggct
acaacgtctc cagatcaacc acagaggatt tcccgctcag gctggagttg 240gctgctccct
cccagacatc tgtgtacttc tgtgccagca gt
282154285DNAArtificial SequenceSynthetic DNA TRBV6-6*04 154aatgctggtg
tcactcagac cccaaaattc cgcatcctga agataggaca gagcatgaca 60ctgcagtgta
cccaggatat gaaccatgaa tacatgtact ggtatcgaca agacccaggc 120atggggctga
agctgattta ttattcagtt ggtgctggta tcactgataa aggagaagtc 180ccgaatggct
acaatgtctc cagatcaacc acagaggatt tcccgctcag gctggagttg 240gctgctccct
cccagacatc tgtgtacttc tgtgccagca gtcga
285155282DNAArtificial SequenceSynthetic DNA TRBV6-6*05 155aatgctggtg
tcactcagac cccaaaattc cgcatcctga agataggaca gagcatgaca 60ctgcagtgtg
cccaggatat gaaccataac tacatgtact ggtatcgaca agacccaggc 120atggggctga
agctgattta ttattcagtt ggtgctggta tcactgacaa aggagaagtc 180ccgaatggct
acaacgtctc cagatcaacc acagaggatt tcccgctcag gctggagttg 240gctgctgcct
cccagacatc tgtgtacttc tgtgccagca gc
282156287DNAArtificial SequenceSynthetic DNA TRBV6-7*01 156aatgctggtg
tcactcagac cccaaaattc cacgtcctga agacaggaca gagcatgact 60ctgctgtgtg
cccaggatat gaaccatgaa tacatgtatc ggtatcgaca agacccaggc 120aaggggctga
ggctgattta ctactcagtt gctgctgctc tcactgacaa aggagaagtt 180cccaatggct
acaatgtctc cagatcaaac acagaggatt tccccctcaa gctggagtca 240gctgctccct
ctcagacttc tgtttacttc tgtgccagca gttactc
287157284DNAArtificial SequenceSynthetic DNA TRBV6-8*01 157aatgctggtg
tcactcagac cccaaaattc cacatcctga agacaggaca gagcatgaca 60ctgcagtgtg
cccaggatat gaaccatgga tacatgtcct ggtatcgaca agacccaggc 120atggggctga
gactgattta ctactcagct gctgctggta ctactgacaa agaagtcccc 180aatggctaca
atgtctctag attaaacaca gaggatttcc cactcaggct ggtgtcggct 240gctccctccc
agacatctgt gtacttgtgt gccagcagtt actc
284158287DNAArtificial SequenceSynthetic DNA TRBV6-9*01 158aatgctggtg
tcactcagac cccaaaattc cacatcctga agacaggaca gagcatgaca 60ctgcagtgtg
cccaggatat gaaccatgga tacttgtcct ggtatcgaca agacccaggc 120atggggctga
ggcgcattca ttactcagtt gctgctggta tcactgacaa aggagaagtc 180cccgatggct
acaatgtatc cagatcaaac acagaggatt tcccgctcag gctggagtca 240gctgctccct
cccagacatc tgtatacttc tgtgccagca gttattc
287159290DNAArtificial SequenceSynthetic DNA TRBV7-1*01 159ggtgctggag
tctcccagtc cctgagacac aaggtagcaa agaagggaaa ggatgtagct 60ctcagatatg
atccaatttc aggtcataat gccctttatt ggtaccgaca gagcctgggg 120cagggcctgg
agtttccaat ttacttccaa ggcaaggatg cagcagacaa atcggggctt 180ccccgtgatc
ggttctctgc acagaggtct gagggatcca tctccactct gaagttccag 240cgcacacagc
agggggactt ggctgtgtat ctctgtgcca gcagctcagc
290160290DNAArtificial SequenceSynthetic DNA TRBV7-2*01 160ggagctggag
tctcccagtc ccccagtaac aaggtcacag agaagggaaa ggatgtagag 60ctcaggtgtg
atccaatttc aggtcatact gccctttact ggtaccgaca gagcctgggg 120cagggcctgg
agtttttaat ttacttccaa ggcaacagtg caccagacaa atcagggctg 180cccagtgatc
gcttctctgc agagaggact gggggatccg tctccactct gacgatccag 240cgcacacagc
aggaggactc ggccgtgtat ctctgtgcca gcagcttagc
290161290DNAArtificial SequenceSynthetic DNA TRBV7-2*02 161ggagctggag
tctcccagtc ccccagtaac aaggtcacag agaagggaaa ggatgtagag 60ctcaggtgtg
atccaatttc aggtcatact gccctttact ggtaccgaca gaggctgggg 120cagggcctgg
agtttttaat ttacttccaa ggcaacagtg caccagacaa atcagggctg 180cccagtgatc
gcttctctgc agagaggact ggggaatccg tctccactct gacgatccag 240cgcacacagc
aggaggactc ggccgtgtat ctctgtgcca gcagcttagc
290162290DNAArtificial SequenceSynthetic DNA TRBV7-2*03 162ggagctggag
tctcccagtc ccccagtaac aaggtcacag agaagggaaa ggatgtagag 60ctcaggtgtg
atccaatttc aggtcatact gccctttact ggtaccgaca gaggctgggg 120cagggcctgg
agtttttaat ttacttccaa ggcaacagtg caccagacaa atcagggctg 180cccagtgatc
gcttctctgc agagaggact ggggaatccg tctccactct gacgatccag 240cgcacacagc
aggaggactc ggccgtgtat ctctgtacca gcagcttagc
290163288DNAArtificial SequenceSynthetic DNA TRBV7-2*04 163ggagctggag
tttcccagtc ccccagtaac aaggtcacag agaagggaaa ggatgtagag 60ctcaggtgtg
atccaatttc aggtcatact gccctttact ggtaccgaca gagcctgggg 120cagggcctgg
agtttttaat ttacttccaa ggcaacagtg caccagacaa atcagggctg 180cccagtgatc
gcttctctgc agagaggact gggggatccg tctccactct gacgatccag 240cgcacacagc
aggaggactc ggccgtgtat ctctgtgcca gcagctta
288164290DNAArtificial SequenceSynthetic DNA TRBV7-3*01 164ggtgctggag
tctcccagac ccccagtaac aaggtcacag agaagggaaa atatgtagag 60ctcaggtgtg
atccaatttc aggtcatact gccctttact ggtaccgaca aagcctgggg 120cagggcccag
agtttctaat ttacttccaa ggcacgggtg cggcagatga ctcagggctg 180cccaacgatc
ggttctttgc agtcaggcct gagggatccg tctctactct gaagatccag 240cgcacagagc
ggggggactc agccgtgtat ctctgtgcca gcagcttaac
290165290DNAArtificial SequenceSynthetic DNA TRBV7-3*02 165ggtgctggag
tctcccagac ccccagtaac aaggtcacag agaagggaaa agatgtagag 60ctcaggtgtg
atccaatttc aggtcatact gccctttact ggtaccgaca aagcctgggg 120cagggcccag
agtttctaat ttacttccaa ggcacgggtg cggcagatga ctcagggctg 180cccaaagatc
ggttctttgc agtcaggcct gagggatccg tctctactct gaagatccag 240cgcacagagc
agggggactc agccgtgtat ctccgtgcca gcagcttaac
290166288DNAArtificial SequenceSynthetic DNA TRBV7-3*03 166ggtgctggag
tctcccagac ccccagtaac aaggtcacag agaagggaaa agatgtagag 60ctcaggtgtg
atccaatttc aggtcatact gccctttact ggtaccgaca aagcctgggg 120cagggcccag
agtttctaat ttacttccaa ggcacgggtg cggcagatga ctcagggctg 180cccaaagatc
ggttctttgc agtcaggcct gagggatccg tctctactct gaagatccag 240cgcacagagc
agggggactc agccgcgtat ctccgtgcca gcagctta
288167285DNAArtificial SequenceSynthetic DNA TRBV7-3*04 167ggtgctggag
tctcccagac ccccagtaac aaggtcacag agaagggaaa atatgtagag 60ctcaggtgtg
atccaatttc aggtcatact gccctttact ggtaccgaca aagcctgggg 120cagggcccag
agtttctaat ttacttccaa ggcacgggtg cggcagatga ctcagggctg 180cccaacgatc
ggttctttgc agtcaggcct gagggatccg tctctactct gaagatccag 240cgcacagagc
ggggggactc tgccgtgtat ctctgtgcca gcagc
285168231DNAArtificial SequenceSynthetic DNA TRBV7-3*05 168tgggagctca
ggtgtgatcc aatttcaggt catactgccc tttactggta ccgacaaagc 60ctggggcagg
gcccagagct tctaatttac ttccaaggca cgggtgcggc agatgactca 120gggctgccca
acgatcggtt ctttgcagtc aggcctgagg gatccgtctc tactctgaag 180atccagcgca
cagagcgggg ggactcagcc gtgtatctct gtgccagcag c
231169290DNAArtificial SequenceSynthetic DNA TRBV7-4*01 169ggtgctggag
tctcccagtc cccaaggtac aaagtcgcaa agaggggacg ggatgtagct 60ctcaggtgtg
attcaatttc gggtcatgta accctttatt ggtaccgaca gaccctgggg 120cagggctcag
aggttctgac ttactcccag agtgatgctc aacgagacaa atcagggcgg 180cccagtggtc
ggttctctgc agagaggcct gagagatccg tctccactct gaagatccag 240cgcacagagc
agggggactc agctgtgtat ctctgtgcca gcagcttagc
290170288DNAArtificial SequenceSynthetic DNA TRBV7-5*01 170ggtgctggag
tctcccagtc cccaaggtac gaagtcacac agaggggaca ggatgtagct 60cccaggtgtg
atccaatttc gggtcaggta accctttatt ggtaccgaca gaccctgggg 120cagggccaag
agtttctgac ttccttccag gatgaaactc aacaagataa atcagggctg 180ctcagtgatc
aattctccac agagaggtct gaggatcttt ctccacctga agatccagcg 240cacagagcaa
gggcgactcg gctgtgtatc tctgtgccag aagcttag
288171289DNAArtificial SequenceSynthetic DNA TRBV7-5*02 171ggtgctggag
tctcccagtc cccaaggtac gaagtcacac agaggggaca ggatgtagct 60cccaggtgtg
atccaatttc gggtcaggta accctttatt ggtaccgaca gaccctgggg 120cagggccaag
agtttctgac ttccttccag gatgaaactc aacaagataa atcagggctg 180ctcagtgatc
aattctccac agagaggtct gaggatcttt ctccacctga agatccagcg 240cacagagcaa
gggcgactcg gctgtgtatc tctgtgtcag aagcttagc
289172290DNAArtificial SequenceSynthetic DNA TRBV7-6*01 172ggtgctggag
tctcccagtc tcccaggtac aaagtcacaa agaggggaca ggatgtagct 60ctcaggtgtg
atccaatttc gggtcatgta tccctttatt ggtaccgaca ggccctgggg 120cagggcccag
agtttctgac ttacttcaat tatgaagccc aacaagacaa atcagggctg 180cccaatgatc
ggttctctgc agagaggcct gagggatcca tctccactct gacgatccag 240cgcacagagc
agcgggactc ggccatgtat cgctgtgcca gcagcttagc
290173285DNAArtificial SequenceSynthetic DNA TRBV7-6*02 173ggtgctggag
tctcccagtc tcccaggtac aaagtcacaa agaggggaca ggatgtagct 60ctcaggtgtg
atccaatctc gggtcatgta tccctttatt ggtaccgaca ggccctgggg 120cagggcccag
agtttctgac ttacttcaat tatgaagccc aacaagacaa atcagggctg 180cccaatgatc
ggttctctgc agagaggcct gagggatcca tctccactct gacgatccag 240cgcacagagc
agcgggactc ggccatgtat cgctgtgcca gcagc
285174290DNAArtificial SequenceSynthetic DNA TRBV7-7*01 174ggtgctggag
tctcccagtc tcccaggtac aaagtcacaa agaggggaca ggatgtaact 60ctcaggtgtg
atccaatttc gagtcatgca accctttatt ggtatcaaca ggccctgggg 120cagggcccag
agtttctgac ttacttcaat tatgaagctc aaccagacaa atcagggctg 180cccagtgatc
ggttctctgc agagaggcct gagggatcca tctccactct gacgattcag 240cgcacagagc
agcgggactc agccatgtat cgctgtgcca gcagcttagc
290175285DNAArtificial SequenceSynthetic DNA TRBV7-7*02 175ggtgctggag
tctcccagtc tcccaggtac aaagtcacaa agaggggaca ggatgtaact 60ctcaggtgtg
atccaatttc gagtcatgta accctttatt ggtatcaaca ggccctgggg 120cagggcccag
agtttctgac ttacttcaat tatgaagctc aaccagacaa atcagggctg 180cccagtgatc
ggttctctgc agagaggcct gagggatcca tctccactct gacgattcag 240cgcacagagc
agcgggactc agccatgtat cgctgtgcca gcagc
285176290DNAArtificial SequenceSynthetic DNA TRBV7-8*01 176ggtgctggag
tctcccagtc ccctaggtac aaagtcgcaa agagaggaca ggatgtagct 60ctcaggtgtg
atccaatttc gggtcatgta tccctttttt ggtaccaaca ggccctgggg 120caggggccag
agtttctgac ttatttccag aatgaagctc aactagacaa atcggggctg 180cccagtgatc
gcttctttgc agaaaggcct gagggatccg tctccactct gaagatccag 240cgcacacagc
aggaggactc cgccgtgtat ctctgtgcca gcagcttagc
290177290DNAArtificial SequenceSynthetic DNA TRBV7-8*02 177ggtgctggag
tctcccagtc ccctaggtac aaagtcgcaa agagaggaca ggatgtagct 60ctcaggtgtg
atccaatttc gggtcatgta tccctttttt ggtaccaaca ggccctgggg 120caggggccag
agtttctgac ttatttccag aatgaagctc aactagacaa atcggggctg 180cccagtgatc
gcttctttgc agaaaggcct gagggatccg tctccactct gaagatccag 240cgcacacaga
aggaggactc cgccgtgtat ctctgtgcca gcagcttagc
290178288DNAArtificial SequenceSynthetic DNA TRBV7-8*03 178ggtgctggag
tctcccagtc ccctaggtac aaagtcgcaa agagaggaca ggatgtagct 60ctcaggtgtg
atccaatttc gggtcatgta tccctttttt ggtaccaaca ggccctcggg 120caggggccag
agtttctgac ttatttccag aatgaagctc aactagacaa atcggggctg 180cccagtgatc
gcttctttgc agaaaggcct gagggatccg tctccactct gaagatccag 240cgcacacagc
aggaggactc cgccgtgtat ctctgtgcca gcagccga
288179288DNAArtificial SequenceSynthetic DNA TRBV7-9*05 179gatactggag
tctcccagaa ccccagacac aagatcacaa agaggggaca gaatgtaact 60ttcaggtgtg
atccaatttc tgaacacaac cgcctttatt ggtaccgaca gaccctgggg 120cagggcccag
agtttctgac ttacttccag aatgaagctc aactagaaaa atcaaggctg 180ctcagtgatc
ggttctctgc agagaggcct aagggatctc tctccacctt ggagatccag 240cgcacagagc
agggggactc ggccatgtat ctctgtgcca gcaccaaa
288180288DNAArtificial SequenceSynthetic DNA TRBV7-9*06 180gatactggag
tctcccagaa ccccagacac aagatcacaa agaggggaca gaatgtaact 60ttcaggtgtg
atccaatttc tgaacacaac cgcctttatt ggtaccgaca gaccctgggg 120cagggcccag
agtttctgac ttacttccag aatgaagctc aactagaaaa atcaaggctg 180ctcagtgatc
ggttctctgc agagaggcct aagggatctc tttccacctt ggagatccag 240cgcacagagc
agggggactc ggccatgtat ctctgtgcca gcacgttg
288181285DNAArtificial SequenceSynthetic DNA TRBV7-9*03 181gatactggag
tctcccagga ccccagacac aagatcacaa agaggggaca gaatgtaact 60ttcaggtgtg
atccaatttc tgaacacaac cgcctttatt ggtaccgaca gaccctgggg 120cagggcccag
agtttctgac ttacttccag aatgaagctc aactagaaaa atcaaggctg 180ctcagtgatc
ggttctctgc agagaggcct aagggatctt tctccacctt ggagatccag 240cgcacagagc
agggggactc ggccatgtat ctctgtgcca gcagc
285182290DNAArtificial SequenceSynthetic DNA TRBV7-9*01 182gatactggag
tctcccagaa ccccagacac aagatcacaa agaggggaca gaatgtaact 60ttcaggtgtg
atccaatttc tgaacacaac cgcctttatt ggtaccgaca gaccctgggg 120cagggcccag
agtttctgac ttacttccag aatgaagctc aactagaaaa atcaaggctg 180ctcagtgatc
ggttctctgc agagaggcct aagggatctt tctccacctt ggagatccag 240cgcacagagc
agggggactc ggccatgtat ctctgtgcca gcagcttagc
290183288DNAArtificial SequenceSynthetic DNA TRBV7-9*02 183gatactggag
tctcccagaa ccccagacac aacatcacaa agaggggaca gaatgtaact 60ttcaggtgtg
atccaatttc tgaacacaac cgcctttatt ggtaccgaca gaccctgggg 120cagggcccag
agtttctgac ttacttccag aatgaagctc aactagaaaa atcaaggctg 180ctcagtgatc
ggttctctgc agagaggcct aagggatctt tctccacctt ggagatccag 240cgcacagagc
agggggactc ggccatgtat ctctgtgcca gcagctta
288184207DNAArtificial SequenceSynthetic DNA TRBV7-9*07 184cacaaccgcc
tttattggta ccgacagacc ctggggcagg gcccagagtt tctgacttac 60ttccagaatg
aagctcaact agaaaaatca aggctgctca gtgatcggtt ctctgcagag 120aggcctaagg
gatctttctc caccttggag atccagcgca cagaggaggg ggactcggcc 180atgtatctct
gtgccagcag cagcagt
207185288DNAArtificial SequenceSynthetic DNA TRBV7-9*04 185atatctggag
tctcccacaa ccccagacac aagatcacaa agaggggaca gaatgtaact 60ttcaggtgtg
atccaatttc tgaacacaac cgcctttatt ggtaccgaca gaaccctggg 120cagggcccag
agtttctgac ttacttccag aatgaagctc aactggaaaa atcagggctg 180ctcagtgatc
ggatctctgc agagaggcct aagggatctt tctccacctt ggagatccag 240cgcacagagc
agggggactc ggccatgtat ctctgtgcca gcagctct
288186279DNAArtificial SequenceSynthetic DNA TRBV8-1*01 186gaggcaggga
tcagccagat accaagatat cacagacaca cagggaaaaa gatcatcctg 60aaatatgctc
agattaggaa ccattattca gtgttctgtt atcaataaga ccaagaatag 120gggctgaggc
tgatccatta ttcaggtagt attggcagca tgaccaaagg cggtgccaag 180gaagggtaca
atgtctctgg aaacaagctc aagcattttc cctcaaccct ggagtctact 240agcaccagcc
agacctctgt acctctgtgg cagtgcatc
279187271DNAArtificial SequenceSynthetic DNA TRBV8-2*01 187gatgctggga
tcacccagat gccaagatat cacattgtac agaagaaaga gatgatcctg 60gaatgtgctc
aggttaggaa cagtgttctg atatcgacag gacccaagac gggggctgaa 120gcttatccac
tattcaggca gtggtcacag caggaccaaa gttgatgtca cagaggggta 180ctgtgtttct
tgaaacaagc ttgagcattt ccccaatcct ggcatccacc agcaccagcc 240agacctatct
gtaccactgt ggcagcacat c
271188286DNAArtificial SequenceSynthetic DNA TRBV9*01 188gattctggag
tcacacaaac cccaaagcac ctgatcacag caactggaca gcgagtgacg 60ctgagatgct
cccctaggtc tggagacctc tctgtgtact ggtaccaaca gagcctggac 120cagggcctcc
agttcctcat tcagtattat aatggagaag agagagcaaa aggaaacatt 180cttgaacgat
tctccgcaca acagttccct gacttgcact ctgaactaaa cctgagctct 240ctggagctgg
gggactcagc tttgtatttc tgtgccagca gcgtag
286189282DNAArtificial SequenceSynthetic DNA TRBV9*03 189gattctggag
tcacacaaac cccaaagcac ctgatcacag caactggaca gcgagtgacg 60ctgagatgct
cccctaggtc tggagacctc tctgtgtact ggtaccaaca gagcctggac 120cagggcctcc
agttcctcat tcaatattat aatggagaag agagagcaaa aggaaacatt 180cttgaacgat
tctccgcaca acagttccct gacttgcact ctgaactaaa cctgagctct 240ctggagctgg
gggactcagc tttgtatttc tgtgccagca gc
282190286DNAArtificial SequenceSynthetic DNA TRBV9*02 190gattctggag
tcacacaaac cccaaagcac ctgatcacag caactggaca gcgagtgacg 60ctgagatgct
cccctaggtc tggagacctc tctgtgtact ggtaccaaca gagcctggac 120cagggcctcc
agttcctcat tcactattat aatggagaag agagagcaaa aggaaacatt 180cttgaacgat
tctccgcaca acagttccct gacttgcact ctgaactaaa cctgagctct 240ctggagctgg
gggactcagc tttgtatttc tgtgccagca gcgtag
286191287DNAArtificial SequenceSynthetic DNA TRBV10-1*01 191gatgctgaaa
tcacccagag cccaagacac aagatcacag agacaggaag gcaggtgacc 60ttggcgtgtc
accagacttg gaaccacaac aatatgttct ggtatcgaca agacctggga 120catgggctga
ggctgatcca ttactcatat ggtgttcaag acactaacaa aggagaagtc 180tcagatggct
acagtgtctc tagatcaaac acagaggacc tccccctcac tctggagtct 240gctgcctcct
cccagacatc tgtatatttc tgcgccagca gtgagtc
287192282DNAArtificial SequenceSynthetic DNA TRBV10-1*02 192gatgctgaaa
tcacccagag cccaagacac aagatcacag agacaggaag gcaggtgacc 60ttggcgtgtc
accagacttg gaaccacaac aatatgttct ggtatcgaca agacctggga 120catgggctga
ggctgatcca ttactcatat ggtgttcacg acactaacaa aggagaagtc 180tcagatggct
acagtgtctc tagatcaaac acagaggacc tccccctcac tctggagtct 240gctgcctcct
cccagacatc tgtatatttc tgcgccagca gt
282193287DNAArtificial SequenceSynthetic DNA TRBV10-2*01 193gatgctggaa
tcacccagag cccaagatac aagatcacag agacaggaag gcaggtgacc 60ttgatgtgtc
accagacttg gagccacagc tatatgttct ggtatcgaca agacctggga 120catgggctga
ggctgatcta ttactcagca gctgctgata ttacagataa aggagaagtc 180cccgatggct
atgttgtctc cagatccaag acagagaatt tccccctcac tctggagtca 240gctacccgct
cccagacatc tgtgtatttc tgcgccagca gtgagtc
287194217DNAArtificial SequenceSynthetic DNA TRBV10-2*02 194aaggcaggtg
accttgatgt gtcaccagac ttggagccac agctatatgt tctggtatcg 60acaagacctg
ggacatgggc tgaggctgat ctattactca gcagctgctg atattacaga 120taaaggagaa
gtccccgatg gctacgttgt ctccagatcc aagacagaga atttccccct 180cactctggag
tcagctaccc gctcccagac atctgtg
217195273DNAArtificial SequenceSynthetic DNA TRBV10-3*03 195gatgctggaa
tcacccagag cccaagacac aaggtcacag agacaggaac accagtgact 60ctgagatgtc
accagactga gaaccaccgc tacatgtact ggtatcgaca agacccgggg 120catgggctga
ggctaatcca ttactcatat ggtgttaaag atactgacaa aggagaagtc 180tcagatggct
atagtgtctc tagatcaaag acagaggatt tcctcctcac tctggagtcc 240gctaccagct
cccagacatc tgtgtacttc tgt
273196273DNAArtificial SequenceSynthetic DNA TRBV10-3*04 196gatgctggaa
tcacccagag cccaagacac aaggtcacag agacaggaac accagtgact 60ctgagatgtc
accagactga gaaccaccgc tacatgtact ggtatcgaca agacccgggg 120catgggctga
ggctgatcca ttactcatat ggtgttaaag atactgacaa aggagaagtc 180tcagatggct
atagtgtctc tagatcaaag acagaggatt tcctcctcac tctggagtcc 240gctaccagct
cccagacatc tgtgtacttc tgt
273197287DNAArtificial SequenceSynthetic DNA TRBV10-3*01 197gatgctggaa
tcacccagag cccaagacac aaggtcacag agacaggaac accagtgact 60ctgagatgtc
accagactga gaaccaccgc tatatgtact ggtatcgaca agacccgggg 120catgggctga
ggctgatcca ttactcatat ggtgttaaag atactgacaa aggagaagtc 180tcagatggct
atagtgtctc tagatcaaag acagaggatt tcctcctcac tctggagtcc 240gctaccagct
cccagacatc tgtgtacttc tgtgccatca gtgagtc
287198287DNAArtificial SequenceSynthetic DNA TRBV10-3*02 198gatgctggaa
tcacccagag cccaagacac aaggtcacag agacaggaac accagtgact 60ctgagatgtc
atcagactga gaaccaccgc tatatgtact ggtatcgaca agacccgggg 120catgggctga
ggctgatcca ttactcatat ggtgttaaag atactgacaa aggagaagtc 180tcagatggct
atagtgtctc tagatcaaag acagaggatt tcctcctcac tctggagtcc 240gctaccagct
cccagacatc tgtgtacttc tgtgccatca gtgagtc
287199290DNAArtificial SequenceSynthetic DNA TRBV11-1*01 199gaagctgaag
ttgcccagtc ccccagatat aagattacag agaaaagcca ggctgtggct 60ttttggtgtg
atcctatttc tggccatgct accctttact ggtaccggca gatcctggga 120cagggcccgg
agcttctggt tcaatttcag gatgagagtg tagtagatga ttcacagttg 180cctaaggatc
gattttctgc agagaggctc aaaggagtag actccactct caagatccag 240cctgcagagc
ttggggactc ggccatgtat ctctgtgcca gcagcttagc
290200290DNAArtificial SequenceSynthetic DNA TRBV11-3*01 200gaagctggag
tggttcagtc tcccagatat aagattatag agaaaaaaca gcctgtggct 60ttttggtgca
atcctatttc tggccacaat accctttact ggtacctgca gaacttggga 120cagggcccgg
agcttctgat tcgatatgag aatgaggaag cagtagacga ttcacagttg 180cctaaggatc
gattttctgc agagaggctc aaaggagtag actccactct caagatccag 240cctgcagagc
ttggggactc ggccgtgtat ctctgtgcca gcagcttaga
290201285DNAArtificial SequenceSynthetic DNA TRBV11-3*02 201gaagctggag
tggttcagtc tcccagatat aagattatag agaaaaagca gcctgtggct 60ttttggtgca
atcctatttc tggccacaat accctttact ggtaccggca gaacttggga 120cagggcccgg
agcttctgat tcgatatgag aatgaggaag cagtagacga ttcacagttg 180cctaaggatc
gattttctgc agagaggctc aaaggagtag actccactct caagatccag 240cctgcagagc
ttggggactc ggccgtgtat ctctgtgcca gcagc
285202269DNAArtificial SequenceSynthetic DNA TRBV11-3*03 202ggtctcccag
atataagatt atagagaaga aacagcctgt ggctttttgg tgcaatccaa 60tttctggcca
caataccctt tactggtacc tgcagaactt gggacagggc ccggagcttc 120tgattcgata
tgagaatgag gaagcagtag acgattcaca gttgcctaag gatcgatttt 180ctgcagagag
gctcaaagga gtagactcca ctctcaagat ccagccagca gagcttgggg 240actcggccat
gtatctctgt gccagcagc
269203290DNAArtificial SequenceSynthetic DNA TRBV11-2*01 203gaagctggag
ttgcccagtc tcccagatat aagattatag agaaaaggca gagtgtggct 60ttttggtgca
atcctatatc tggccatgct accctttact ggtaccagca gatcctggga 120cagggcccaa
agcttctgat tcagtttcag aataacggtg tagtggatga ttcacagttg 180cctaaggatc
gattttctgc agagaggctc aaaggagtag actccactct caagatccag 240cctgcaaagc
ttgaggactc ggccgtgtat ctctgtgcca gcagcttaga
290204285DNAArtificial SequenceSynthetic DNA TRBV11-2*03 204gaagctggag
ttgcccagtc tcccagatat aagattatag agaaaaggca gagtgtggct 60ttttggtgca
atcctatatc tggccatgct accctttact ggtaccagca gatcctggga 120cagggcccaa
agcttctgat tcagtttcag aataacggtg tagtggatga ttcacagttg 180cctaaggatc
gattttctgc agagaggctc aaaggagtag actccactct caagatccaa 240cctgcaaagc
ttgaggactc ggccgtgtat ctctgtgcca gcagc
285205285DNAArtificial SequenceSynthetic DNA TRBV11-2*02 205gaagctggag
ttgcccagtc tcccagatat aagattatag agaaaaggca gagtgtggct 60ttttggtgca
atcctatatc tggccatgct accctttact ggtaccagca gatcctggga 120cagggcccaa
agcttctgat tcagtttcag aataacggtg tagtggatga ttcacagttg 180cctaaggatc
gattttctgc agagaggctc aaaggagtag actccactct caagatccag 240cctgcaaagc
ttgagaactc ggccgtgtat ctctgtgcca gcagt
285206290DNAArtificial SequenceSynthetic DNA TRBV12-1*01 206gatgctggtg
ttatccagtc acccaggcac aaagtgacag agatgggaca atcagtaact 60ctgagatgcg
aaccaatttc aggccacaat gatcttctct ggtacagaca gacctttgtg 120cagggactgg
aattgctgaa ttacttctgc agctggaccc tcgtagatga ctcaggagtg 180tccaaggatt
gattctcagc acagatgcct gatgtatcat tctccactct gaggatccag 240cccatggaac
ccagggactt gggcctatat ttctgtgcca gcagctttgc
290207290DNAArtificial SequenceSynthetic DNA TRBV12-2*01 207gatgctggca
ttatccagtc acccaagcat gaggtgacag aaatgggaca aacagtgact 60ctgagatgtg
agccaatttt tggccacaat ttccttttct ggtacagaga taccttcgtg 120cagggactgg
aattgctgag ttacttccgg agctgatcta ttatagataa tgcaggtatg 180cccacagagc
gattctcagc tgagaggcct gatggatcat tctctactct gaagatccag 240cctgcagagc
agggggactc ggccgtgtat gtctgtgcaa gtcgcttagc
290208290DNAArtificial SequenceSynthetic DNA TRBV12-4*01 208gatgctggag
ttatccagtc accccggcac gaggtgacag agatgggaca agaagtgact 60ctgagatgta
aaccaatttc aggacacgac taccttttct ggtacagaca gaccatgatg 120cggggactgg
agttgctcat ttactttaac aacaacgttc cgatagatga ttcagggatg 180cccgaggatc
gattctcagc taagatgcct aatgcatcat tctccactct gaagatccag 240ccctcagaac
ccagggactc agctgtgtac ttctgtgcca gcagtttagc
290209288DNAArtificial SequenceSynthetic DNA TRBV12-4*02 209gatgctggag
ttatccagtc accccggcac gaggtgacag agatgggaca agaagtgact 60ctgagatgta
aaccaatttc aggacatgac taccttttct ggtacagaca gaccatgatg 120cggggactgg
agttgctcat ttactttaac aacaacgttc cgatagatga ttcagggatg 180cccgaggatc
gattctcagc taagatgcct aatgcatcat tctccactct gaggatccag 240ccctcagaac
ccagggactc agctgtgtac ttctgtgcca gcagttta
288210290DNAArtificial SequenceSynthetic DNA TRBV12-3*01 210gatgctggag
ttatccagtc accccgccat gaggtgacag agatgggaca agaagtgact 60ctgagatgta
aaccaatttc aggccacaac tcccttttct ggtacagaca gaccatgatg 120cggggactgg
agttgctcat ttactttaac aacaacgttc cgatagatga ttcagggatg 180cccgaggatc
gattctcagc taagatgcct aatgcatcat tctccactct gaagatccag 240ccctcagaac
ccagggactc agctgtgtac ttctgtgcca gcagtttagc
290211290DNAArtificial SequenceSynthetic DNA TRBV12-5*01 211gatgctagag
tcacccagac accaaggcac aaggtgacag agatgggaca agaagtaaca 60atgagatgtc
agccaatttt aggccacaat actgttttct ggtacagaca gaccatgatg 120caaggactgg
agttgctggc ttacttccgc aaccgggctc ctctagatga ttcggggatg 180ccgaaggatc
gattctcagc agagatgcct gatgcaactt tagccactct gaagatccag 240ccctcagaac
ccagggactc agctgtgtat ttttgtgcta gtggtttggt
290212287DNAArtificial SequenceSynthetic DNA TRBV13*01 212gctgctggag
tcatccagtc cccaagacat ctgatcaaag aaaagaggga aacagccact 60ctgaaatgct
atcctatccc tagacacgac actgtctact ggtaccagca gggtccaggt 120caggaccccc
agttcctcat ttcgttttat gaaaagatgc agagcgataa aggaagcatc 180cctgatcgat
tctcagctca acagttcagt gactatcatt ctgaactgaa catgagctcc 240ttggagctgg
gggactcagc cctgtacttc tgtgccagca gcttagg
287213282DNAArtificial SequenceSynthetic DNA TRBV13*02 213gctgctggag
tcatccagtc cccaagacat ctgatcagag aaaagaggga aacagccact 60ctgaaatgct
atcctatccc tagacacgac actgtctact ggtaccagca gggcccaggt 120caggaccccc
agttcttcat ttcgttttat gaaaagatgc agagcgataa aggaagcatc 180cctgatcgat
tctcagctca acagttcagt gactatcatt ctgaactgaa catgagctcc 240ttggagctgg
gggactcagc cctgtacttc tgtgccagca gc
282214290DNAArtificial SequenceSynthetic DNA TRBV14*01 214gaagctggag
ttactcagtt ccccagccac agcgtaatag agaagggcca gactgtgact 60ctgagatgtg
acccaatttc tggacatgat aatctttatt ggtatcgacg tgttatggga 120aaagaaataa
aatttctgtt acattttgtg aaagagtcta aacaggatga gtccggtatg 180cccaacaatc
gattcttagc tgaaaggact ggagggacgt attctactct gaaggtgcag 240cctgcagaac
tggaggattc tggagtttat ttctgtgcca gcagccaaga
290215285DNAArtificial SequenceSynthetic DNA TRBV14*02 215gaagctggag
ttactcagtt ccccagccac agcgtaatag agaagggcca gactgtgact 60ctgagatgtg
acccaatttc tggacatgat aatctttatt ggtatcgacg tgttatggga 120aaagaaataa
aatttctgtt acattttgtg aaagagtcta aacaggatga atccggtatg 180cccaacaatc
gattcttagc tgaaaggact ggagggacgt attctactct gaaggtgcag 240cctgcagaac
tggaggattc tggagtttat ttctgtgcca gcagc
285216287DNAArtificial SequenceSynthetic DNA TRBV15*01 216gatgccatgg
tcatccagaa cccaagatac caggttaccc agtttggaaa gccagtgacc 60ctgagttgtt
ctcagacttt gaaccataac gtcatgtact ggtaccagca gaagtcaagt 120caggccccaa
agctgctgtt ccactactat gacaaagatt ttaacaatga agcagacacc 180cctgataact
tccaatccag gaggccgaac acttctttct gctttcttga catccgctca 240ccaggcctgg
gggacacagc catgtacctg tgtgccacca gcagaga
287217282DNAArtificial SequenceSynthetic DNA TRBV15*03 217gatgccatgg
tcatccagaa cccaagatac cgggttaccc agtttggaaa gccagtgacc 60ctgagttgtt
ctcagacttt gaaccataac gtcatgtact ggtaccagca gaagtcaagt 120caggccccaa
agctgctgtt ccactactat aacaaagatt ttaacaatga agcagacacc 180cctgataact
tccaatccag gaggccgaac acttctttct gctttctaga catccgctca 240ccaggcctgg
gggacgcagc catgtaccag tgtgccacca gc
282218282DNAArtificial SequenceSynthetic DNA TRBV15*02 218gatgccatgg
tcatccagaa cccaagatac caggttaccc agtttggaaa gccagtgacc 60ctgagttgtt
ctcagacttt gaaccataac gtcatgtact ggtaccagca gaagtcaagt 120caggccccaa
agctgctgtt ccactactat gacaaagatt ttaacaatga agcagacacc 180cctgataact
tccaatccag gaggccgaac acttctttct gctttcttga catccgctca 240ccaggcctgg
gggacgcagc catgtacctg tgtgccacca gc
282219290DNAArtificial SequenceSynthetic DNA TRBV16*01 219ggtgaagaag
tcgcccagac tccaaaacat cttgtcagag gggaaggaca gaaagcaaaa 60ttatattgtg
ccccaataaa aggacacagt tatgtttttt ggtaccaaca ggtcctgaaa 120aacgagttca
agttcttgat ttccttccag aatgaaaatg tctttgatga aacaggtatg 180cccaaggaaa
gattttcagc taagtgcctc ccaaattcac cctgtagcct tgagatccag 240gctacgaagc
ttgaggattc agcagtgtat ttttgtgcca gcagccaatc
290220290DNAArtificial SequenceSynthetic DNA TRBV16*02 220ggtgaagaag
tcgcccagac tccaaaacat cttgtcagag gggaaggaca gaaagcaaaa 60ttatattgtg
ccccaataaa aggacacagt taggtttttt ggtaccaaca ggtcctgaaa 120aacgagttca
agttcttgat ttccttccag aatgaaaatg tctttgatga aacaggtatg 180cccaaggaaa
gattttcagc taagtgcctc ccaaattcac cctgtagcct tgagatccag 240gctacgaagc
ttgaggattc agcagtgtat ttttgtgcca gcagccaatc
290221285DNAArtificial SequenceSynthetic DNA TRBV16*03 221ggtgaagaag
tcgcccagac tccaaaacat cttgtcagag gggaaggaca gaaagcaaaa 60ttatattgtg
ccccaataaa aggacacagt tatgtttttt ggtaccaaca ggtcctgaaa 120aacgagttca
agttcttggt ttccttccag aatgaaaatg tctttgatga aacaggtatg 180cccaaggaaa
gattttcagc taagtgcctc ccaaattcac cctgtagcct tgagatccag 240gctacgaagc
ttgaggattc agcagtgtat ttttgtgcca gcagc
285222287DNAArtificial SequenceSynthetic DNA TRBV17*01 222gagcctggag
tcagccagac ccccagacac aaggtcacca acatgggaca ggaggtgatt 60ctgaggtgcg
atccatcttc tggtcacatg tttgttcact ggtaccgaca gaatctgagg 120caagaaatga
agttgctgat ttccttccag taccaaaaca ttgcagttga ttcagggatg 180cccaaggaac
gattcacagc tgaaagacct aacggaacgt cttccacgct gaagatccat 240cccgcagagc
cgagggactc agccgtgtat ctctacagta gcggtgg
287223290DNAArtificial SequenceSynthetic DNA TRBV18*01 223aatgccggcg
tcatgcagaa cccaagacac ctggtcagga ggaggggaca ggaggcaaga 60ctgagatgca
gcccaatgaa aggacacagt catgtttact ggtatcggca gctcccagag 120gaaggtctga
aattcatggt ttatctccag aaagaaaata tcatagatga gtcaggaatg 180ccaaaggaac
gattttctgc tgaatttccc aaagagggcc ccagcatcct gaggatccag 240caggtagtgc
gaggagattc ggcagcttat ttctgtgcca gctcaccacc
290224287DNAArtificial SequenceSynthetic DNA TRBV19*01 224gatggtggaa
tcactcagtc cccaaagtac ctgttcagaa aggaaggaca gaatgtgacc 60ctgagttgtg
aacagaattt gaaccacgat gccatgtact ggtaccgaca ggacccaggg 120caagggctga
gattgatcta ctactcacag atagtaaatg actttcagaa aggagatata 180gctgaagggt
acagcgtctc tcgggagaag aaggaatcct ttcctctcac tgtgacatcg 240gcccaaaaga
acccgacagc tttctatctc tgtgccagta gtataga
287225287DNAArtificial SequenceSynthetic DNA TRBV19*02 225gatggtggaa
tcactcagtc cccaaagtac ctgttcagaa aggaaggaca gaatgtgacc 60ctgagttgtg
aacagaattt gaaccacgat gccatgtact ggtaccgaca ggtcccaggg 120caagggctga
gattgatcta ctactcacac atagtaaatg actttcagaa aggagatata 180gctgaagggt
acagcgtctc tcgggagaag aaggaatcct ttcctctcac tgtgacatcg 240gcccaaaaga
acccgacagc tttctatctc tgtgccagta gtataga
287226282DNAArtificial SequenceSynthetic DNA TRBV19*03 226gatggtggaa
tcactcagtc cccaaagtac ctgttcagaa aggaaggaca gaatgtgacc 60ctgagttgtg
aacagaattt gaaccacgat gccatgtact ggtaccgaca ggacccaggg 120caagggctga
gattgatcta ctactcacac atagtaaatg actttcagaa aggagatata 180gctgaagggt
acagcgtctc tcgggagaag aaggaatcct ttcctctcac tgtgacatcg 240gcccaaaaga
acccgacagc tttctatctc tgtgccagta gc
282227291DNAArtificial SequenceSynthetic DNA TRBV20-1*05 227ggtgctgtcg
tctctcaaca tccgagcagg gttatctgta agagtggaac ctctgtgaag 60atcgagtgcc
gttccctgga ctttcaggcc acaactatgt tttggtatcg tcagttcccg 120aaaaagagtc
tcatgctgat ggcaacttcc aatgagggct ccaaggccac atacgagcaa 180ggcgtcgaga
aggacaagtt tctcatcaac catgcaagcc tgaccttgtc cactctgaca 240gtgaccagtg
cccatcctga agacagcagc ttctacatct gcagtgctag a
291228291DNAArtificial SequenceSynthetic DNA TRBV20-1*07 228ggtgctgtcg
tctctcaaca tccgagcagg gttatctgta agagtggaac ctctgtgaag 60atcgagtgcc
gttccctgga ctttcaggcc acaactatgt tttggtatcg tcagttcccg 120aaaaagagtc
tcatgcagat cgcaacttcc aatgagggct ccaaggccac atacgagcaa 180ggcgtcgaga
aggacaagtt tctcatcaac catgcaagcc tgaccttgtc cactctgaca 240gtgaccagtg
cccatcctga agacagcagc ttctacatct gcagtgctag a
291229291DNAArtificial SequenceSynthetic DNA TRBV20-1*04 229ggtgctgtcg
tctctcaaca tccgagcagg gttatctgta agagtggaac ctctgtgaag 60atcgagtgcc
gttccttgga ctttcaggcc acaactatgt tttggtatcg tcagttcccg 120aaaaagagtc
tcatgctgat ggcaacttcc aatgagggct ccaaggccac atacgagcaa 180ggcgtcgaga
aggacaagtt tctcatcaac catgcaagcc tgaccttgtc cactctgaca 240gtgaccagtg
cccatcctga agacagcagc ttctacatct gcagtgctag t
291230288DNAArtificial SequenceSynthetic DNA TRBV20-1*06 230ggtgctgtcg
tctctcaaca tccgagtagg gttatctgta agagtggaac ctctgtgaag 60atcgagtgcc
gttccctgga ctttcaggcc acaactatgt tttggtatcg tcagttcccg 120aaaaagagtc
tcatgctgat ggcaacttcc aatgagggct ccaaggccac atacgagcaa 180ggcgtcgaga
aggacaagtt tctcatcaac catgcaagcc tgaccttgtc cactctgaca 240gtgaccagtg
cccatcctga agacagcagc ttctacatct gcagtgct
288231288DNAArtificial SequenceSynthetic DNA TRBV20-1*02 231ggtgctgtcg
tctctcaaca tccgagcagg gttatctgta agagtggaac ctctgtgaag 60atcgagtgcc
gttccctgga ctttcaggcc acaactatgt tttggtatcg tcagttcccg 120aaacagagtc
tcatgctgat ggcaacttcc aatgagggct ccaaggccac atacgagcaa 180ggcgtcgaga
aggacaagtt tctcatcaac catgcaagcc tgaccttgtc cactctgaca 240gtgaccagtg
cccatcctga agacagcagc ttctacatct gcagtgct
288232293DNAArtificial SequenceSynthetic DNA TRBV20-1*01 232ggtgctgtcg
tctctcaaca tccgagctgg gttatctgta agagtggaac ctctgtgaag 60atcgagtgcc
gttccctgga ctttcaggcc acaactatgt tttggtatcg tcagttcccg 120aaacagagtc
tcatgctgat ggcaacttcc aatgagggct ccaaggccac atacgagcaa 180ggcgtcgaga
aggacaagtt tctcatcaac catgcaagcc tgaccttgtc cactctgaca 240gtgaccagtg
cccatcctga agacagcagc ttctacatct gcagtgctag aga
293233288DNAArtificial SequenceSynthetic DNA TRBV20-1*03 233ggtgctgtcg
tctctcaaca tccgagctgg gttatctgta agagtggaac ctctgtgaag 60atcgagtgcc
gttccctgga ctttcaggcc acaactatgt tttggtatcg tcagttcccg 120aaacagagtc
tcatgctgat ggcaacttcc aatgagggct gcaaggccac atacgagcaa 180ggcgtcgaga
aggacaagtt tctcatcaac catgcaagcc tgaccttgtc cactctgaca 240gtgaccagtg
cccatcctga agacagcagc ttctacatct gcagtgct
288234290DNAArtificial SequenceSynthetic DNA TRBV21-1*01 234gacaccaagg
tcacccagag acctagactt ctggtcaaag caagtgaaca gaaagcaaag 60atggattgtg
ttcctataaa agcacatagt tatgtttact ggtatcgtaa gaagctggaa 120gaagagctca
agtttttggt ttactttcag aatgaagaac ttattcagaa agcagaaata 180atcaatgagc
gatttttagc ccaatgctcc aaaaactcat cctgtacctt ggagatccag 240tccacggagt
caggggacac agcactgtat ttctgtgcca gcagcaaagc
290235288DNAArtificial SequenceSynthetic DNA TRBV22-1*01 235gatgctgaca
tctatcagat gccattccag ctcactgggg ctggatggga tgtgactctg 60gagtggaaac
ggaatttgag acacaatgac atgtactgct actggtactg gcaggaccca 120aagcaaaatc
tgagactgat ctattactca agggttgaaa aggatattca gagaggagat 180ctaactgaag
gctacgtgtc tgccaagagg agaaggggct atttcttctc agggtgaagt 240tggcccacac
cagccaaaca gctttgtact tctgtcctgg gagcgcac
288236290DNAArtificial SequenceSynthetic DNA TRBV23-1*01 236catgccaaag
tcacacagac tccaggacat ttggtcaaag gaaaaggaca gaaaacaaag 60atggattgta
cccccgaaaa aggacatact tttgtttatt ggtatcaaca gaatcagaat 120aaagagttta
tgcttttgat ttcctttcag aatgaacaag ttcttcaaga aacggagatg 180cacaagaagc
gattctcatc tcaatgcccc aagaacgcac cctgcagcct ggcaatcctg 240tcctcagaac
cgggagacac ggcactgtat ctctgcgcca gcagtcaatc
290237288DNAArtificial SequenceSynthetic DNA TRBV24-1*01 237gatgctgatg
ttacccagac cccaaggaat aggatcacaa agacaggaaa gaggattatg 60ctggaatgtt
ctcagactaa gggtcatgat agaatgtact ggtatcgaca agacccagga 120ctgggcctac
ggttgatcta ttactccttt gatgtcaaag atataaacaa aggagagatc 180tctgatggat
acagtgtctc tcgacaggca caggctaaat tctccctgtc cctagagtct 240gccatcccca
accagacagc tctttacttc tgtgccacca gtgatttg
288238287DNAArtificial SequenceSynthetic DNA TRBV25-1*01 238gaagctgaca
tctaccagac cccaagatac cttgttatag ggacaggaaa gaagatcact 60ctggaatgtt
ctcaaaccat gggccatgac aaaatgtact ggtatcaaca agatccagga 120atggaactac
acctcatcca ctattcctat ggagttaatt ccacagagaa gggagatctt 180tcctctgagt
caacagtctc cagaataagg acggagcatt ttcccctgac cctggagtct 240gccaggccct
cacatacctc tcagtacctc tgtgccagca gtgaata
287239287DNAArtificial SequenceSynthetic DNA TRBV26*01 239gatgctgtag
ttacacaatt cccaagacac agaatcattg ggacaggaaa ggaattcatt 60ctacagtgtt
cccagaatat gaatcatgtt acaatgtact ggtatcgaca ggacccagga 120cttggactga
agctggtcta ttattcacct ggcactggga gcactgaaaa aggagatatc 180tctgaggggt
atcatgtttc ttgaaatact atagcatctt ttcccctgac cctgaagtct 240gccagcacca
accagacatc tgtgtatctc tatgccagca gttcatc
287240287DNAArtificial SequenceSynthetic DNA TRBV27*01 240gaagcccaag
tgacccagaa cccaagatac ctcatcacag tgactggaaa gaagttaaca 60gtgacttgtt
ctcagaatat gaaccatgag tatatgtcct ggtatcgaca agacccaggg 120ctgggcttaa
ggcagatcta ctattcaatg aatgttgagg tgactgataa gggagatgtt 180cctgaagggt
acaaagtctc tcgaaaagag aagaggaatt tccccctgat cctggagtcg 240cccagcccca
accagacctc tctgtacttc tgtgccagca gtttatc
287241287DNAArtificial SequenceSynthetic DNA TRBV28*01 241gatgtgaaag
taacccagag ctcgagatat ctagtcaaaa ggacgggaga gaaagttttt 60ctggaatgtg
tccaggatat ggaccatgaa aatatgttct ggtatcgaca agacccaggt 120ctggggctac
ggctgatcta tttctcatat gatgttaaaa tgaaagaaaa aggagatatt 180cctgaggggt
acagtgtctc tagagagaag aaggagcgct tctccctgat tctggagtcc 240gccagcacca
accagacatc tatgtacctc tgtgccagca gtttatg
287242290DNAArtificial SequenceSynthetic DNA TRBV29-1*01 242agtgctgtca
tctctcaaaa gccaagcagg gatatctgtc aacgtggaac ctccctgacg 60atccagtgtc
aagtcgatag ccaagtcacc atgatgttct ggtaccgtca gcaacctgga 120cagagcctga
cactgatcgc aactgcaaat cagggctctg aggccacata tgagagtgga 180tttgtcattg
acaagtttcc catcagccgc ccaaacctaa cattctcaac tctgactgtg 240agcaacatga
gccctgaaga cagcagcata tatctctgca gcgttgaaga
290243288DNAArtificial SequenceSynthetic DNA TRBV29-1*02 243agtgctgtca
tctctcaaaa gccaagcagg gatatctgtc aacgtggaac ctccctgacg 60atccagtgtc
aagtcgatag ccaagtcacc atgatgttct ggtaccgtca gcaacctgga 120cagagcctga
cactgatcgc aactgcaaat cagggctctg aggccacata tgagagtgga 180tttgtcattg
acaagtttcc catcagccgc ccaaacctaa cattctcaag tctgactgtg 240agcaacatga
gccctgaaga cagcagcata tatctctgca gcgttgaa
288244231DNAArtificial SequenceSynthetic DNA TRBV29-1*03 244acgatccagt
gtcaagtcga tagccaagtc accatgatat tctggtaccg tcagcaacct 60ggacagagcc
tgacactgat cgcaactgca aatcagggct ctgaggccac atatgagagt 120ggatttgtca
ttgacaagtt tcccatcagc cgcccaaacc taacattctc aactctgact 180gtgagcaaca
tgagccctga agacagcagc atatatctct gcagcgcggg c
231245284DNAArtificial SequenceSynthetic DNA TRBV30*02 245tctcagacta
ttcatcaatg gccagcgacc ctggtgcagc ctgtgggcag cccgctctct 60ctggagtgca
ctgtggaggg aacatcaaac cccaacctat actggtaccg acaggctgca 120ggcaggggcc
tccagctgct cttctactcc gttggtattg gccagatcag ctctgaggtg 180ccccagaatc
tctcagcctc cagaccccag gaccggcagt tcatcctgag ttctaagaag 240ctcctcctca
gtgactctgg cttctatctc tgtgcctgga gtgt
284246282DNAArtificial SequenceSynthetic DNA TRBV30*05 246tctcagacta
ttcatcaatg gccagcgacc ctggtgcagc ctgtgggcag cccgctctcc 60ctggagtgca
ctgtggaggg aacatcaaac cccaacctat actggtaccg acaggctgca 120ggacggggcc
tccagctgct cttctactcc gttggtattg gccagatcag ctctgaggtg 180ccccagaatc
tctcagcctc cagaccccag gaccggcagt tcatcctgag ttctaagaag 240ctccttctca
gtgactctgg cttctatctc tgtgcctggg ga
282247284DNAArtificial SequenceSynthetic DNA TRBV30*01 247tctcagacta
ttcatcaatg gccagcgacc ctggtgcagc ctgtgggcag cccgctctct 60ctggagtgca
ctgtggaggg aacatcaaac cccaacctat actggtaccg acaggctgca 120ggcaggggcc
tccagctgct cttctactcc gttggtattg gccagatcag ctctgaggtg 180ccccagaatc
tctcagcctc cagaccccag gaccggcagt tcatcctgag ttctaagaag 240ctccttctca
gtgactctgg cttctatctc tgtgcctgga gtgt
284248276DNAArtificial SequenceSynthetic DNA TRBV30*04 248actattcatc
aatggccagc gaccctggtg cagcctgtgg gcagcccgct ctctctggag 60tgcactgtgg
agggaacatc aaaccccaac ctatactggt accgacaggc tgcaggcagg 120ggcctccagc
tgctcttcta ctccattggt attgaccaga tcagctctga ggtgccccag 180aatctctcag
cctccagacc ccaggaccgg cagttcattc tgagttctaa gaagctcctc 240ctcagtgact
ctggcttcta tctctgtgcc tggagt
276249448DNAArtificial SequenceSynthetic DNA TCRBJ1S1 249ttgaaaaagg
aacctaggac cctgtggatg gactctgtca ttctccatgg tcctaaaaag 60caaaagtcaa
agtgttcttc tgtgtaatac ccataaagca caggaggaga tttcttagct 120cactgtcctc
catcctagcc agggccctct cccctctcta tgccttcaat gtgattttca 180ccttgacccc
tgtcactgtg tgaacactga agctttcttt ggacaaggca ccagactcac 240agttgtaggt
aagacatttt tcaggttctt ttgcagatcc gtcacaggga aaagtgggtc 300cacagtgtcc
cttttagagt ggctatattc ttatgtgcta actatggcta caccttcggt 360tcggggacca
ggttaaccgt tgtaggtaag gctgggggtc tctaggaggg gtgcgatgag 420ggaggactct
gtcctgggaa atgtcaaa
448250448DNAArtificial SequenceSynthetic DNA TCRBJ1S2 250gccagggccc
tctcccctct ctatgccttc aatgtgattt tcaccttgac ccctgtcact 60gtgtgaacac
tgaagctttc tttggacaag gcaccagact cacagttgta ggtaagacat 120ttttcaggtt
cttttgcaga tccgtcacag ggaaaagtgg gtccacagtg tcccttttag 180agtggctata
ttcttatgtg ctaactatgg ctacaccttc ggttcgggga ccaggttaac 240cgttgtaggt
aaggctgggg gtctctagga ggggtgcgat gagggaggac tctgtcctgg 300gaaatgtcaa
agagaacaga gatcccagct cccggagcca gactgaggga gacgtcatgt 360catgtcccgg
gattgagttc aggggaggct ccctgtgagg gcgaatccac ccaggcttcc 420cagaggctct
gagcagtcac agctgagc
448251450DNAArtificial SequenceSynthetic DNA TCRBJ1S3 251gattttatag
gaggccactc tgtgtctctt tttgtcacct gcctgagtct tgggcaagct 60ctggaaggga
acacagagta ctggaagcag agctgctgtc cctgtgaggg aagagttccc 120atgaactccc
aacctctgcc tgaatcccag ctgtgctcag cagagactgg ggggttttga 180agtggccctg
ggaggctgtg ctctggaaac accatatatt ttggagaggg aagttggctc 240actgttgtag
gtgagtaagt caaggctgga cagctgggaa cttgcaaaaa ggggctggaa 300tccagacgga
gcctttgtct ctagtgctta ggtgaaagtg tatttttgtc aggaaggcct 360atgaggcaga
tgaggagggg atagcctccc tctcctctcg actattttgt agactgcctg 420tgccaagtta
ggttccccta ctgagagatg
450252451DNAArtificial SequenceSynthetic DNA TCRBJ1S4 252cagaagaggg
aacttggggg atcacacggg gcctaattgg tctgctgacc accgcatttt 60gggttgtacc
attgtctacc cctctaccca ccagggttaa aattctacta aggaacagga 120gaggacctgg
caggtggact tggggaggca ggagtggaag gcagcaggtc gcggttttcc 180ttccagtctt
taatgttgtg caactaatga aaaactgttt tttggcagtg gaacccagct 240ctctgtcttg
ggtatgtaaa agacttcttt cgggatagtg tatcataagg tcggagttcc 300aggaggaccc
cttgcgggag ggcagaaact gagaacacag ccaagaaaag ctcataaaat 360gtgggtcagt
ggagtgtgtg gtggggcccc aagagttctg tgtgtaagca gcttctggaa 420ggaagggccc
acaccagctc ctctggggtt t
451253450DNAArtificial SequenceSynthetic DNA TCRBJ1S5 253gatagtgtat
cataaggtcg gagttccagg aggacccctt gcgggagggc agaaactgag 60aacacagcca
agaaaagctc ataaaatgtg ggtcagtgga gtgtgtggtg gggccccaag 120agttctgtgt
gtaagcagct tctggaagga agggcccaca ccagctcctc tggggtttgc 180cacactcatg
atgcactgtg tagcaatcag ccccagcatt ttggtgatgg gactcgactc 240tccatcctag
gtaagttgca gaatcagggt ggtatggcca ttgtcccttg aaggcagagt 300tctctgcttc
tcctcccggt gctggtgagg cagattgagt aaaatctctt accccatggg 360gtaagagctg
tgcctgtgcc tgcgttccct ttggtgtgtc ttggttgact cctctatttc 420tcttctctaa
gtcttcagtc cataatctgc
450254453DNAArtificial SequenceSynthetic DNA TCRBJ1S6 254atggctctgc
ctctcctaag cctcttcctc ttgcgcctta tgctgcacag tatgcttagg 60cctttttcct
aacagaatcc ctttggtcca gagccatgaa tccaggcaga gaaaggcagc 120catcctgctg
tcagggagct aagacttgcc ctctgactgg agatcgccgg gtgggtttta 180tctaagcctc
tgcagctgtg ctcctataat tcacccctcc actttgggaa cgggaccagg 240ctcactgtga
caggtatggg ggctccactc ttgactcggg ggtgcctggg tttgactgca 300atgatcagtt
gctgggaagg gaattgagtg taagaacgga ggtcagggtc accccttctt 360acctggagca
ctgtgccctc tcctcccctc cctggagctc ttccagcttg ttgctctgct 420gtgttgcctg
cagttcctca gctgtagagc tcc
453255449DNAArtificial SequenceSynthetic DNA TCRBJ2S1 255aatccactgt
gttgtccccc agccaagtgg attctcctct gcaaattggt ggtggcctca 60tgcaagatcc
agttaccgtg tccagctaac tcgagacagg aaaagatagg ctcaggaaag 120agaggaaggg
tgtgccctct gtctgtgcta agggaggtgg ggaaggagaa ggaattctgg 180gcagcccctt
cccactgtgc tcctacaatg agcagttctt cgggccaggg acacggctca 240ccgtgctagg
taagaagggg gctccaggtg ggagagaggg tgagcagccc agcctgcacg 300accccagaac
cctgttctta ggggagtgga cactgggcaa tccagggccc tcctcgaggg 360aagcggggtt
tgcgccaggg tccccagggc tgtgcgaaca ccggggagct gttttttgga 420gaaggctcta
ggctgaccgt actgggtaa
449256451DNAArtificial SequenceSynthetic DNA TCRBJ2S2 256ctgtgctcct
acaatgagca gttcttcggg ccagggacac ggctcaccgt gctaggtaag 60aagggggctc
caggtgggag agagggtgag cagcccagcc tgcacgaccc cagaaccctg 120ttcttagggg
agtggacact gggcaatcca gggccctcct cgagggaagc ggggtttgcg 180ccagggtccc
cagggctgtg cgaacaccgg ggagctgttt tttggagaag gctctaggct 240gaccgtactg
ggtaaggagg cggttggggc tccggagagc tccgagaggg cgggatgggc 300agaggtaagc
agctgcccca ctctgagagg ggctgtgctg agaggcgctg ctgggcgtct 360gggcggagga
ctcctggttc tgggtgctgg gagagcgatg gggctctcag cggtgggaag 420gacccgagct
gagtctggga cagcagagcg g
451257449DNAArtificial SequenceSynthetic DNA TCRBJ2S3 257gggcgggatg
ggcagaggta agcagctgcc ccactctgag aggggctgtg ctgagaggcg 60ctgctgggcg
tctgggcgga ggactcctgg ttctgggtgc tgggagagcg atggggctct 120cagcggtggg
aaggacccga gctgagtctg ggacagcaga gcgggcagca ccggtttttg 180tcctgggcct
ccaggctgtg agcacagata cgcagtattt tggcccaggc acccggctga 240cagtgctcgg
taagcggggg ctcccgctga agccccggaa ctggggaggg ggcgccccgg 300gacgccgggg
gcgtcgcagg gccagtttct gtgccgcgtc tcggggctgt gagccaaaaa 360cattcagtac
ttcggcgccg ggacccggct ctcagtgctg ggtaagctgg ggccgccggg 420ggaccgggga
cgagactgcg ctcgggttt
449258450DNAArtificial SequenceSynthetic DNA TCRBJ2S4 258gacagcagag
cgggcagcac cggtttttgt cctgggcctc caggctgtga gcacagatac 60gcagtatttt
ggcccaggca cccggctgac agtgctcggt aagcgggggc tcccgctgaa 120gccccggaac
tggggagggg gcgccccggg acgccggggg cgtcgcaggg ccagtttctg 180tgccgcgtct
cggggctgtg agccaaaaac attcagtact tcggcgccgg gacccggctc 240tcagtgctgg
gtaagctggg gccgccgggg gaccggggac gagactgcgc tcgggttttt 300gtgcggggct
cgggggccgt gaccaagaga cccagtactt cgggccaggc acgcggctcc 360tggtgctcgg
tgagcgcggg ctgctggggc gcgggcgcgg gcggcttggg tctggttttt 420gcggggagtc
cccgggctgt gctctggggc
450259448DNAArtificial SequenceSynthetic DNA TCRBJ2S5 259ccccggaact
ggggaggggg cgccccggga cgccgggggc gtcgcagggc cagtttctgt 60gccgcgtctc
ggggctgtga gccaaaaaca ttcagtactt cggcgccggg acccggctct 120cagtgctggg
taagctgggg ccgccggggg accggggacg agactgcgct cgggtttttg 180tgcggggctc
gggggccgtg accaagagac ccagtacttc gggccaggca cgcggctcct 240ggtgctcggt
gagcgcgggc tgctggggcg cgggcgcggg cggcttgggt ctggtttttg 300cggggagtcc
ccgggctgtg ctctggggcc aacgtcctga ctttcggggc cggcagcagg 360ctgaccgtgc
tgggtgagtt ttcgcgggac cacccgggcg gcgggattca ggtggaaggc 420ggcggctgct
tcgcggcacc cggtccgg
448260453DNAArtificial SequenceSynthetic DNA TCRBJ2S6 260cagtgctggg
taagctgggg ccgccggggg accggggacg agactgcgct cgggtttttg 60tgcggggctc
gggggccgtg accaagagac ccagtacttc gggccaggca cgcggctcct 120ggtgctcggt
gagcgcgggc tgctggggcg cgggcgcggg cggcttgggt ctggtttttg 180cggggagtcc
ccgggctgtg ctctggggcc aacgtcctga ctttcggggc cggcagcagg 240ctgaccgtgc
tgggtgagtt ttcgcgggac cacccgggcg gcgggattca ggtggaaggc 300ggcggctgct
tcgcggcacc cggtccggcc ctgtgctggg agacctgggc tgggtcccca 360gggtgggcag
gagctcgggg agccttagag gtttgcatgc gggggtgcac ctccgtgctc 420ctacgagcag
tacttcgggc cgggcaccag gct
453261447DNAArtificial SequenceSynthetic DNA TCRBJ2S7 261tgactttcgg
ggccggcagc aggctgaccg tgctgggtga gttttcgcgg gaccacccgg 60gcggcgggat
tcaggtggaa ggcggcggct gcttcgcggc acccggtccg gccctgtgct 120gggagacctg
ggctgggtcc ccagggtggg caggagctcg gggagcctta gaggtttgca 180tgcgggggtg
cacctccgtg ctcctacgag cagtacttcg ggccgggcac caggctcacg 240gtcacaggtg
agattcgggc gtctccccac cttccagccc ctcggtcccc ggagtcggag 300ggtggaccgg
agctggagga gctgggtgtc cggggtcagc tctgcaaggt cacctccccg 360ctcctgggga
aagactgggg aagagggagg gggtggggag gtgctcagag tccggaaagc 420tgagcagagg
gcgaggccac ttttaat
447262296DNAArtificial SequenceSynthetic DNA IGHV1-46*02 262caggtgcagc
tggtgcagtc tggggctgag gtgaagaagc ctggggcctc agtgaaggtt 60tcctgcaagg
catctggata caccttcaac agctactata tgcactgggt gcgacaggcc 120cctggacaag
ggcttgagtg gatgggaata atcaacccta gtggtggtag cacaagctac 180gcacagaagt
tccagggcag agtcaccatg accagggaca cgtccacgag cacagtctac 240atggagctga
gcagcctgag atctgaggac acggccgtgt attactgtgc gagaga
296263260DNAArtificial SequenceSynthetic DNA IGHV1/OR15-5*01
263agaagcctgg ggcctcagtg aaggtctcct gcaaggcttc tggatacacc ttcaccagct
60actgtatgca ctgggtgcac caggtccatg cacaagggct tgagtggatg ggattggtgt
120gccctagtga tggcagcaca agctatgcac agaagttcca ggccagagtc accataacca
180gggacacatc catgagcaca gcctacatgg agctaagcag tctgagatct gaggacacgg
240ccatgtatta ctgtgtgaga
260264294DNAArtificial SequenceSynthetic DNA IGHV1/OR15-5*03
264caggtacagc tggtgcagtc tggggctgag gtgaagaagc ctggggcctc agtgaaggtc
60tcctgcaagg cttctggata caccttcacc aactactgta tgcactgggt gcgccaggtc
120catgcacaag ggcttgagtg gatgggattg gtgtgcccta gtgatggcag cacaagctat
180gcacaaaagt tccaggccag agtcaccata accagggaca catccatgag cacagcctac
240atggagctaa gcagtctgag atctgaggac acggccatgt attactgtgt gaga
294265296DNAArtificial SequenceSynthetic DNA IGHV1/OR15-9*01
265caggtacagc tgatgcagtc tggggctgag gtgaagaagc ctggggcctc agtgaggatc
60tcctgcaagg cttctggata caccttcacc agctactgta tgcactgggt gtgccaggcc
120catgcacaag ggcttgagtg gatgggattg gtgtgcccta gtgatggcag cacaagctat
180gcacagaagt tccagggcag agtcaccata accagggaca catccatggg cacagcctac
240atggagctaa gcagcctgag atctgaggac acggccatgt attactgtgt gagaga
296266260DNAArtificial SequenceSynthetic DNA IGHV1-c*01 266ggaagtctgg
ggcctcagtg aaagtctcct gtagtttttc tgggtttacc atcaccagct 60acggtataca
ttgggtgcaa cagtcccctg gacaagggct tgagtggatg ggatggatca 120accctggcaa
tggtagccca agctatgcca agaagtttca gggcagattc accatgacca 180gggacatgtc
cacaaccaca gcctacacag acctgagcag cctgacatct gaggacatgg 240ctgtgtatta
ctatgcaaga
260267296DNAArtificial SequenceSynthetic DNA IGHV1-NL1*01 267caggttcagc
tgttgcagcc tggggtccag gtgaagaagc ctgggtcctc agtgaaggtc 60tcctgctagg
cttccagata caccttcacc aaatacttta cacggtgggt gtgacaaagc 120cctggacaag
ggcatnagtg gatgggatga atcaaccctt acaacgataa cacacactac 180gcacagacgt
tctggggcag agtcaccatt accagtgaca ggtccatgag cacagcctac 240atggagctga
gcngcctgag atccgaagac atggtcgtgt attactgtgt gagaga
296268296DNAArtificial SequenceSynthetic DNA IGHV1-58*01 268caaatgcagc
tggtgcagtc tgggcctgag gtgaagaagc ctgggacctc agtgaaggtc 60tcctgcaagg
cttctggatt cacctttact agctctgctg tgcagtgggt gcgacaggct 120cgtggacaac
gccttgagtg gataggatgg atcgtcgttg gcagtggtaa cacaaactac 180gcacagaagt
tccaggaaag agtcaccatt accagggaca tgtccacaag cacagcctac 240atggagctga
gcagcctgag atccgaggac acggccgtgt attactgtgc ggcaga
296269296DNAArtificial SequenceSynthetic DNA IGHV1-58*02 269caaatgcagc
tggtgcagtc tgggcctgag gtgaagaagc ctgggacctc agtgaaggtc 60tcctgcaagg
cttctggatt cacctttact agctctgcta tgcagtgggt gcgacaggct 120cgtggacaac
gccttgagtg gataggatgg atcgtcgttg gcagtggtaa cacaaactac 180gcacagaagt
tccaggaaag agtcaccatt accagggaca tgtccacaag cacagcctac 240atggagctga
gcagcctgag atccgaggac acggccgtgt attactgtgc ggcaga
296270275DNAArtificial SequenceSynthetic DNA IGHV1-69*03 270caggtgcagc
tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60tcctgcaagg
cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120cctggacaag
ggcttgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180gcacagaagt
tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240atggagctga
gcagcctgag atctgatgac acggc
275271233DNAArtificial SequenceSynthetic DNA IGHV1-69*07 271agaagcctgg
gtcctcggtg aaggtctcct gcaaggcttc tggaggcacc ttcagcagct 60atgctatcag
ctgggtgcga caggcccctg gacaagggct tgagtggatg ggaaggatca 120tccctatctt
tggtacagca aactacgcac agaagttcca gggcagagtc acgattaccg 180cggacgaatc
cacgagcaca gcctacatgg agctgagcag cctgagatct gag
233272296DNAArtificial SequenceSynthetic DNA IGHV1-69*12 272caggtccagc
tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60tcctgcaagg
cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120cctggacaag
ggcttgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180gcacagaagt
tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240atggagctga
gcagcctgag atctgaggac acggccgtgt attactgtgc gagaga
296273294DNAArtificial SequenceSynthetic DNA IGHV1-69*05 273caggtccagc
tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60tcctgcaagg
cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120cctggacaag
ggcttgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180gcacagaagt
tccagggcag agtcacgatt accacggacg aatccacgag cacagcctac 240atggagctga
gcagcctgag atctgaggac acggccgtgt attactgtgc gaga
294274296DNAArtificial SequenceSynthetic DNA IGHV1-69*13 274caggtccagc
tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc agtgaaggtc 60tcctgcaagg
cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120cctggacaag
ggcttgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180gcacagaagt
tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240atggagctga
gcagcctgag atctgaggac acggccgtgt attactgtgc gagaga
296275296DNAArtificial SequenceSynthetic DNA IGHV1-69*01 275caggtgcagc
tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60tcctgcaagg
cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120cctggacaag
ggcttgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180gcacagaagt
tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240atggagctga
gcagcctgag atctgaggac acggccgtgt attactgtgc gagaga
296276296DNAArtificial SequenceSynthetic DNA IGHV1-69*06 276caggtgcagc
tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60tcctgcaagg
cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120cctggacaag
ggcttgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180gcacagaagt
tccagggcag agtcacgatt accgcggaca aatccacgag cacagcctac 240atggagctga
gcagcctgag atctgaggac acggccgtgt attactgtgc gagaga
296277294DNAArtificial SequenceSynthetic DNA IGHV1-69*02 277caggtccagc
tggtgcaatc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60tcctgcaagg
cttctggagg caccttcagc agctatacta tcagctgggt gcgacaggcc 120cctggacaag
ggcttgagtg gatgggaagg atcatcccta tccttggtat agcaaactac 180gcacagaagt
tccagggcag agtcacgatt accgcggaca aatccacgag cacagcctac 240atggagctga
gcagcctgag atctgaggac acggccgtgt attactgtgc gaga
294278296DNAArtificial SequenceSynthetic DNA IGHV1-69*08 278caggtccagc
tggtgcaatc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60tcctgcaagg
cttctggagg caccttcagc agctatacta tcagctgggt gcgacaggcc 120cctggacaag
ggcttgagtg gatgggaagg atcatcccta tccttggtac agcaaactac 180gcacagaagt
tccagggcag agtcacgatt accgcggaca aatccacgag cacagcctac 240atggagctga
gcagcctgag atctgaggac acggccgtgt attactgtgc gagaga
296279296DNAArtificial SequenceSynthetic DNA IGHV1-69*04 279caggtccagc
tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60tcctgcaagg
cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120cctggacaag
ggcttgagtg gatgggaagg atcatcccta tccttggtat agcaaactac 180gcacagaagt
tccagggcag agtcacgatt accgcggaca aatccacgag cacagcctac 240atggagctga
gcagcctgag atctgaggac acggccgtgt attactgtgc gagaga
296280296DNAArtificial SequenceSynthetic DNA IGHV1-69*11 280caggtccagc
tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60tcctgcaagg
cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120cctggacaag
ggcttgagtg gatgggaagg atcatcccta tccttggtac agcaaactac 180gcacagaagt
tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac 240atggagctga
gcagcctgag atctgaggac acggccgtgt attactgtgc gagaga
296281296DNAArtificial SequenceSynthetic DNA IGHV1-69*09 281caggtgcagc
tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60tcctgcaagg
cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120cctggacaag
ggcttgagtg gatgggaagg atcatcccta tccttggtat agcaaactac 180gcacagaagt
tccagggcag agtcacgatt accgcggaca aatccacgag cacagcctac 240atggagctga
gcagcctgag atctgaggac acggccgtgt attactgtgc gagaga
296282296DNAArtificial SequenceSynthetic DNA IGHV1-69*10 282caggtccagc
tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc agtgaaggtc 60tcctgcaagg
cttctggagg caccttcagc agctatgcta tcagctgggt gcgacaggcc 120cctggacaag
ggcttgagtg gatgggaggg atcatcccta tccttggtat agcaaactac 180gcacagaagt
tccagggcag agtcacgatt accgcggaca aatccacgag cacagcctac 240atggagctga
gcagcctgag atctgaggac acggccgtgt attactgtgc gagaga
296283294DNAArtificial SequenceSynthetic DNA IGHV1-f*01 283gaggtccagc
tggtacagtc tggggctgag gtgaagaagc ctggggctac agtgaaaatc 60tcctgcaagg
tttctggata caccttcacc gactactaca tgcactgggt gcaacaggcc 120cctggaaaag
ggcttgagtg gatgggactt gttgatcctg aagatggtga aacaatatac 180gcagagaagt
tccagggcag agtcaccata accgcggaca cgtctacaga cacagcctac 240atggagctga
gcagcctgag atctgaggac acggccgtgt attactgtgc aaca
294284233DNAArtificial SequenceSynthetic DNA IGHV1-f*02 284agaagcctgg
ggctacagtg aaaatctcct gcaaggtttc tggatacacc ttcaccgact 60actacatgca
ctgggtgcaa caggcccctg gaaaagggct tgagtggatg ggacttgttg 120atcctgaaga
tggtgaaaca atatatgcag agaagttcca gggcagagtc accataaccg 180cggacacgtc
tacagacaca gcctacatgg agctgagcag cctgagatct gag
233285296DNAArtificial SequenceSynthetic DNA IGHV1-24*01 285caggtccagc
tggtacagtc tggggctgag gtgaagaagc ctggggcctc agtgaaggtc 60tcctgcaagg
tttccggata caccctcact gaattatcca tgcactgggt gcgacaggct 120cctggaaaag
ggcttgagtg gatgggaggt tttgatcctg aagatggtga aacaatctac 180gcacagaagt
tccagggcag agtcaccatg accgaggaca catctacaga cacagcctac 240atggagctga
gcagcctgag atctgaggac acggccgtgt attactgtgc aacaga
296286294DNAArtificial SequenceSynthetic DNA IGHV7-4-1*01 286caggtgcagc
tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt 60tcctgcaagg
cttctggata caccttcact agctatgcta tgaattgggt gcgacaggcc 120cctggacaag
ggcttgagtg gatgggatgg atcaacacca acactgggaa cccaacgtat 180gcccagggct
tcacaggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat 240ctgcagatct
gcagcctaaa ggctgaggac actgccgtgt attactgtgc gaga
294287274DNAArtificial SequenceSynthetic DNA IGHV7-4-1*03 287caggtgcagc
tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt 60tcctgcaagg
cttctggata caccttcact agctatgcta tgaattgggt gcgacaggcc 120cctggacaag
ggcttgagtg gatgggatgg atcaacacca acactgggaa cccaacgtat 180gcccagggct
tcacaggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat 240ctgcagatca
gcacgctaaa ggctgaggac actg
274288296DNAArtificial SequenceSynthetic DNA IGHV7-4-1*02 288caggtgcagc
tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt 60tcctgcaagg
cttctggata caccttcact agctatgcta tgaattgggt gcgacaggcc 120cctggacaag
ggcttgagtg gatgggatgg atcaacacca acactgggaa cccaacgtat 180gcccagggct
tcacaggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat 240ctgcagatca
gcagcctaaa ggctgaggac actgccgtgt attactgtgc gagaga
296289296DNAArtificial SequenceSynthetic DNA IGHV7-81*01 289caggtgcagc
tggtgcagtc tggccatgag gtgaagcagc ctggggcctc agtgaaggtc 60tcctgcaagg
cttctggtta cagtttcacc acctatggta tgaattgggt gccacaggcc 120cctggacaag
ggcttgagtg gatgggatgg ttcaacacct acactgggaa cccaacatat 180gcccagggct
tcacaggacg gtttgtcttc tccatggaca cctctgccag cacagcatac 240ctgcagatca
gcagcctaaa ggctgaggac atggccatgt attactgtgc gagata
296290289DNAArtificial SequenceSynthetic DNA IGHV7-40*03 290ctgcagctgg
tgcagtctgg gcctgaggtg aagaagcctg gggcctcagt gaaggtctcc 60tataagtctt
ctggttacac cttcaccatc tatggtatga attgggtatg atagacccct 120ggacagggct
ttgagtggat gtgatggatc atcacctaca ctgggaaccc aacgtatacc 180cacggcttca
caggatggtt tgtcttctcc atggacacgt ctgtcagcac ggcgtgtctt 240cagatcagca
gcctaaaggc tgaggacacg gccgagtatt actgtgcga
289291296DNAArtificial SequenceSynthetic DNA IGHV5-51*01 291gaggtgcagc
tggtgcagtc tggagcagag gtgaaaaagc ccggggagtc tctgaagatc 60tcctgtaagg
gttctggata cagctttacc agctactgga tcggctgggt gcgccagatg 120cccgggaaag
gcctggagtg gatggggatc atctatcctg gtgactctga taccagatac 180agcccgtcct
tccaaggcca ggtcaccatc tcagccgaca agtccatcag caccgcctac 240ctgcagtgga
gcagcctgaa ggcctcggac accgccatgt attactgtgc gagaca
296292245DNAArtificial SequenceSynthetic DNA IGHV5-51*05 292aaaagcccgg
ggagtctctg aagatctcct gtaagggttc tggatacagc tttaccagct 60actggatcgg
ctgggtgcgc cagatgccca ggaaaggcct ggagtggatg gggatcatct 120atcctggtga
ctctgatacc agatacagcc cgtccttcca aggccaggtc accatctcag 180ccgacaagtc
catcagcacc gcctacctgc agtggagcag cctgaaggcc tcggacaccg 240ccatg
245293296DNAArtificial SequenceSynthetic DNA IGHV5-51*02 293gaggtgcagc
tggtgcagtc tggagcagag gtgaaaaagc ccggggagtc tctgaagatc 60tcctgtaagg
gttctggata cagctttacc agctactgga ccggctgggt gcgccagatg 120cccgggaaag
gcttggagtg gatggggatc atctatcctg gtgactctga taccagatac 180agcccgtcct
tccaaggcca ggtcaccatc tcagccgaca agtccatcag caccgcctac 240ctgcagtgga
gcagcctgaa ggcctcggac accgccatgt attactgtgc gagaca
296294294DNAArtificial SequenceSynthetic DNA IGHV5-51*03 294gaggtgcagc
tggtgcagtc tggagcagag gtgaaaaagc cgggggagtc tctgaagatc 60tcctgtaagg
gttctggata cagctttacc agctactgga tcggctgggt gcgccagatg 120cccgggaaag
gcctggagtg gatggggatc atctatcctg gtgactctga taccagatac 180agcccgtcct
tccaaggcca ggtcaccatc tcagccgaca agtccatcag caccgcctac 240ctgcagtgga
gcagcctgaa ggcctcggac accgccatgt attactgtgc gaga
294295294DNAArtificial SequenceSynthetic DNA IGHV5-51*04 295gaggtgcagc
tggtgcagtc tggagcagag gtgaaaaagc cgggggagtc tctgaagatc 60tcctgtaagg
gttctggata cagctttacc agctactgga tcggctgggt gcgccagatg 120cccgggaaag
gcctggagtg gatggggatc atctatcctg gtgactctga taccagatac 180agcccgtcct
tccaaggcca ggtcaccatc tcagccgaca agcccatcag caccgcctac 240ctgcagtgga
gcagcctgaa ggcctcggac accgccatgt attactgtgc gaga
294296294DNAArtificial SequenceSynthetic DNA IGHV5-a*01 296gaagtgcagc
tggtgcagtc tggagcagag gtgaaaaagc ccggggagtc tctgaggatc 60tcctgtaagg
gttctggata cagctttacc agctactgga tcagctgggt gcgccagatg 120cccgggaaag
gcctggagtg gatggggagg attgatccta gtgactctta taccaactac 180agcccgtcct
tccaaggcca cgtcaccatc tcagctgaca agtccatcag cactgcctac 240ctgcagtgga
gcagcctgaa ggcctcggac accgccatgt attactgtgc gaga
294297294DNAArtificial SequenceSynthetic DNA IGHV5-a*03 297gaagtgcagc
tggtgcagtc cggagcagag gtgaaaaagc ccggggagtc tctgaggatc 60tcctgtaagg
gttctggata cagctttacc agctactgga tcagctgggt gcgccagatg 120cccgggaaag
gcctggagtg gatggggagg attgatccta gtgactctta taccaactac 180agcccgtcct
tccaaggcca cgtcaccatc tcagctgaca agtccatcag cactgcctac 240ctgcagtgga
gcagcctgaa ggcctcggac accgccatgt attactgtgc gaga
294298294DNAArtificial SequenceSynthetic DNA IGHV5-a*04 298gaagtgcagc
tggtgcagtc tggagcagag gtgaaaaagc ccggggagtc tctgaggatc 60tcctgtaagg
gttctggata cagctttacc agctactgga tcagctgggt gcgccagatg 120cccgggaaag
gcctggagtg gatggggagg attgatccta gtgactctta taccaactac 180agcccgtcct
tccaaggcca ggtcaccatc tcagctgaca agtccatcag cactgcctac 240ctgcagtgga
gcagcctgaa ggcctcggac accgccatgt attactgtgc gaga
294299295DNAArtificial SequenceSynthetic DNA IGHV5-a*02 299gaagtgcagc
tggtgcagtc tggagcagag gtgaaaaagc ccggggagtc tctgaggatc 60tcctgtaagg
gttctggata cagctttacc agctactgga tcagctgggt gcgccagatg 120cccgggaaag
gcttggagtg gatggggagg attgatccta gtgactctta taccaactac 180agcccgtcct
tccaaggcca cgtcaccatc tcagctgaca agtccatcag cactgcctac 240ctgcagtgga
gcagcctgaa ggctcggaca ccgccatgta ttactgtgcg agaca
295300294DNAArtificial SequenceSynthetic DNA IGHV5-78*01 300gaggtgcagc
tgttgcagtc tgcagcagag gtgaaaagac ccggggagtc tctgaggatc 60tcctgtaaga
cttctggata cagctttacc agctactgga tccactgggt gcgccagatg 120cccgggaaag
aactggagtg gatggggagc atctatcctg ggaactctga taccagatac 180agcccatcct
tccaaggcca cgtcaccatc tcagccgaca gctccagcag caccgcctac 240ctgcagtgga
gcagcctgaa ggcctcggac gccgccatgt attattgtgt gaga
294301296DNAArtificial SequenceSynthetic DNA IGHV3-11*01 301caggtgcagc
tggtggagtc tgggggaggc ttggtcaagc ctggagggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt gactactaca tgagctggat ccgccaggct 120ccagggaagg
ggctggagtg ggtttcatac attagtagta gtggtagtac catatactac 180gcagactctg
tgaagggccg attcaccatc tccagggaca acgccaagaa ctcactgtat 240ctgcaaatga
acagcctgag agccgaggac acggccgtgt attactgtgc gagaga
296302294DNAArtificial SequenceSynthetic DNA IGHV3-11*03 302caggtgcagc
tgttggagtc tgggggaggc ttggtcaagc ctggagggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt gactactaca tgagctggat ccgccaggct 120ccagggaagg
ggctggagtg ggtttcatac attagtagta gtagtagtta cacaaactac 180gcagactctg
tgaagggccg attcaccatc tccagagaca acgccaagaa ctcactgtat 240ctgcaaatga
acagcctgag agccgaggac acggccgtgt attactgtgc gaga
294303296DNAArtificial SequenceSynthetic DNA IGHV3-21*01 303gaggtgcagc
tggtggagtc tgggggaggc ctggtcaagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatagca tgaactgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtctcatcc attagtagta gtagtagtta catatactac 180gcagactcag
tgaagggccg attcaccatc tccagagaca acgccaagaa ctcactgtat 240ctgcaaatga
acagcctgag agccgaggac acggctgtgt attactgtgc gagaga
296304296DNAArtificial SequenceSynthetic DNA IGHV3-21*02 304gaggtgcaac
tggtggagtc tgggggaggc ctggtcaagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatagca tgaactgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtctcatcc attagtagta gtagtagtta catatactac 180gcagactcag
tgaagggccg attcaccatc tccagagaca acgccaagaa ctcactgtat 240ctgcaaatga
acagcctgag agccgaggac acggctgtgt attactgtgc gagaga
296305296DNAArtificial SequenceSynthetic DNA IGHV3-48*01 305gaggtgcagc
tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatagca tgaactgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtttcatac attagtagta gtagtagtac catatactac 180gcagactctg
tgaagggccg attcaccatc tccagagaca atgccaagaa ctcactgtat 240ctgcaaatga
acagcctgag agccgaggac acggctgtgt attactgtgc gagaga
296306296DNAArtificial SequenceSynthetic DNA IGHV3-48*02 306gaggtgcagc
tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatagca tgaactgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtttcatac attagtagta gtagtagtac catatactac 180gcagactctg
tgaagggccg attcaccatc tccagagaca atgccaagaa ctcactgtat 240ctgcaaatga
acagcctgag agacgaggac acggctgtgt attactgtgc gagaga
296307293DNAArtificial SequenceSynthetic DNA IGHV3-h*01 307gaggtgcagc
tggtggagtc tgggggaggc ttggtaaagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt gactactaca tgaactgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtctcatcc attagtagta gtagtaccat atactacgca 180gactctgtga
agggccgatt caccatctcc agagacaacg ccaagaactc actgtatctg 240caaatgaaca
gcctgagagc cgaggacacg gctgtgtatt actgtgcgag aga
293308293DNAArtificial SequenceSynthetic DNA IGHV3-h*02 308gaggtgcagc
tggtggagtc tgggggaggc ttggtaaagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt gactactaca tgaactgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtctcatcc attagtagta gtagtaccat atactacgca 180gactctgtga
agggccgatt caccatctcc agagacaacg ccaagaactc actgtatctg 240caaatgaaca
gcctgagagc cgaggacacg gctgtttatt actgtgcgag aga
293309296DNAArtificial SequenceSynthetic DNA IGHV3-48*03 309gaggtgcagc
tggtggagtc tgggggaggc ttggtacagc ctggagggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agttatgaaa tgaactgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtttcatac attagtagta gtggtagtac catatactac 180gcagactctg
tgaagggccg attcaccatc tccagagaca acgccaagaa ctcactgtat 240ctgcaaatga
acagcctgag agccgaggac acggctgttt attactgtgc gagaga
296310292DNAArtificial SequenceSynthetic DNA IGHV3/OR16-8*01
310gaggtgcagc tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactg
60tcctgtccag cctctggatt caccttcagt aaccactaca tgagctgggt ccgccaggct
120ccagggaagg gactggagtg ggtttcatac attagtggtg atagtggtta cacaaactac
180gcagactctg tgaagggccg attcaccatc tccagggaca acgccaataa ctcaccgtat
240ctgcaaatga acagcctgag agctgaggac acggctgtgt attactgtgt ga
292311292DNAArtificial SequenceSynthetic DNA IGHV3/OR16-9*01
311gaggtgcagc tggtggagtc tggaggaggc ttggtacagc ctggggggtc cctgagactc
60tcctgtgcag cctctggatt caccttcagt aaccactaca cgagctgggt ccgccaggct
120ccagggaagg gactggagtg ggtttcatac agtagtggta atagtggtta cacaaactac
180gcagactctg tgaaaggccg attcaccatc tccagggaca acgccaagaa ctcactgtat
240ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgt ga
292312293DNAArtificial SequenceSynthetic DNA IGHV3-13*01 312gaggtgcagc
tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctacgaca tgcactgggt ccgccaagct 120acaggaaaag
gtctggagtg ggtctcagct attggtactg ctggtgacac atactatcca 180ggctccgtga
agggccgatt caccatctcc agagaaaatg ccaagaactc cttgtatctt 240caaatgaaca
gcctgagagc cggggacacg gctgtgtatt actgtgcaag aga
293313291DNAArtificial SequenceSynthetic DNA IGHV3-13*03 313gaggtgcagc
tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag
cctgtggatt caccttcagt agctacgaca tgcactgggt ccgccaagct 120acaggaaaag
gtctggagtg ggtctcagct attggtactg ctggtgacac atactatcca 180ggctccgtga
agggccaatt caccatctcc agagaaaatg ccaagaactc cttgtatctt 240caaatgaaca
gcctgagagc cggggacacg gctgtgtatt actgtgcaag a
291314293DNAArtificial SequenceSynthetic DNA IGHV3-13*02 314gaggtgcatc
tggtggagtc tgggggaggc ttggtacagc ctgggggggc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt aactacgaca tgcactgggt ccgccaagct 120acaggaaaag
gtctggagtg ggtctcagcc aatggtactg ctggtgacac atactatcca 180ggctccgtga
aggggcgatt caccatctcc agagaaaatg ccaagaactc cttgtatctt 240caaatgaaca
gcctgagagc cggggacacg gctgtgtatt actgtgcaag aga
293315123DNAArtificial SequenceSynthetic DNA IGHV3-47*02 315atactatgca
gactccgtga tgggccgatt caccatctcc agagacaacg ccaagaagtc 60cttgtatctt
caaatgaaca gcctgatagc tgaggacatg gctgtgtatt attgtgcaag 120aga
123316282DNAArtificial SequenceSynthetic DNA IGHV3-47*03 316gaggatcagc
tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagaccc 60tcctgtgcag
cctctggatt cgccttcagt agctatgttc tgcactgggt tcgccgggct 120ccagggaagg
gtccggagtg ggtatcagct attggtactg gtggtgatac atactatgca 180gactccgtga
tgggccgatt caccatctcc agagacaacg ccaagaagtc cttgtatctc 240aaatgaacag
cctgatagct gaggacatgg ctgtgtatta tg
282317291DNAArtificial SequenceSynthetic DNA IGHV3-47*01 317gaggatcagc
tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgcgaccc 60tcctgtgcag
cctctggatt cgccttcagt agctatgctc tgcactgggt tcgccgggct 120ccagggaagg
gtctggagtg ggtatcagct attggtactg gtggtgatac atactatgca 180gactccgtga
tgggccgatt caccatctcc agagacaacg ccaagaagtc cttgtatctt 240catatgaaca
gcctgatagc tgaggacatg gctgtgtatt attgtgcaag a
291318291DNAArtificial SequenceSynthetic DNA IGHV3/OR16-10*01
318gaggttcagc tggtgcagtc tgggggaggc ttggtacatc ctggggggtc cctgagactc
60tcctgtgcag gctctggatt caccttcagt agctatgcta tgcactgggt tcgccaggct
120ccaggaaaag gtctggagtg ggtatcagct attggtactg gtggtggcac atactatgca
180gactccgtga agggccgatt caccatctcc agagacaatg ccaagaactc cttgtatctt
240caaatgaaca gcctgagagc cgaggacatg gctgtgtatt actgtgcaag a
291319291DNAArtificial SequenceSynthetic DNA IGHV3/OR16-10*02
319gaggttcagc tggtgcagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc
60tcctgtgcag gctctggatt caccttcagt agctatgcta tgcactgggt tcgccaggct
120ccaggaaaag gtctggagtg ggtatcagct attggtactg gtggtggcac atactatgca
180gactccgtga agggccgatt caccatctcc agagacaatg ccaagaactc cttgtatctt
240caaatgaaca gcctgagagc cgaggacatg gctgtgtatt actgtgcaag a
291320296DNAArtificial SequenceSynthetic DNA IGHV3-62*01 320gaggtgcagc
tggtggagtc tggggaaggc ttggtccagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctctgcta tgcactgggt ccgccaggct 120ccaagaaagg
gtttgtagtg ggtctcagtt attagtacaa gtggtgatac cgtactctac 180acagactctg
tgaagggccg attcaccatc tccagagaca atgcccagaa ttcactgtct 240ctgcaaatga
acagcctgag agccgagggc acagttgtgt actactgtgt gaaaga
296321296DNAArtificial SequenceSynthetic DNA IGHV3-64*01 321gaggtgcagc
tggtggagtc tgggggaggc ttggtccagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatgcta tgcactgggt ccgccaggct 120ccagggaagg
gactggaata tgtttcagct attagtagta atgggggtag cacatattat 180gcaaactctg
tgaagggcag attcaccatc tccagagaca attccaagaa cacgctgtat 240cttcaaatgg
gcagcctgag agctgaggac atggctgtgt attactgtgc gagaga
296322296DNAArtificial SequenceSynthetic DNA IGHV3-64*02 322gaggtgcagc
tggtggagtc tggggaaggc ttggtccagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatgcta tgcactgggt ccgccaggct 120ccagggaagg
gactggaata tgtttcagct attagtagta atgggggtag cacatattat 180gcagactctg
tgaagggcag attcaccatc tccagagaca attccaagaa cacgctgtat 240cttcaaatgg
gcagcctgag agctgaggac atggctgtgt attactgtgc gagaga
296323296DNAArtificial SequenceSynthetic DNA IGHV3-64*03 323gaggtgcagc
tggtggagtc tgggggaggc ttggtccagc ctggggggtc cctgagactc 60tcctgttcag
cctctggatt caccttcagt agctatgcta tgcactgggt ccgccaggct 120ccagggaagg
gactggaata tgtttcagct attagtagta atgggggtag cacatactac 180gcagactcag
tgaagggcag attcaccatc tccagagaca attccaagaa cacgctgtat 240gtccaaatga
gcagtctgag agctgaggac acggctgtgt attactgtgt gaaaga
296324296DNAArtificial SequenceSynthetic DNA IGHV3-64*05 324gaggtgcagc
tggtggagtc tgggggaggc ttggtccagc ctggggggtc cctgagactc 60tcctgttcag
cctctggatt caccttcagt agctatgcta tgcactgggt ccgccaggct 120ccagggaagg
gactggaata tgtttcagct attagtagta atgggggtag cacatactac 180gcagactcag
tgaagggcag attcaccatc tccagagaca attccaagaa cacgctgtat 240gttcaaatga
gcagtctgag agctgaggac acggctgtgt attactgtgt gaaaga
296325296DNAArtificial SequenceSynthetic DNA IGHV3-64*04 325caggtgcagc
tggtggagtc tgggggaggc ttggtccagc ctggggggtc cctgagactc 60tcctgttcag
cctctggatt caccttcagt agctatgcta tgcactgggt ccgccaggct 120ccagggaagg
gactggaata tgtttcagct attagtagta atgggggtag cacatactac 180gcagactcag
tgaagggcag attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agctgaggac acggctgtgt attactgtgc gagaga
296326296DNAArtificial SequenceSynthetic DNA IGHV3-16*01 326gaggtacaac
tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt aacagtgaca tgaactgggc ccgcaaggct 120ccaggaaagg
ggctggagtg ggtatcgggt gttagttgga atggcagtag gacgcactat 180gtggactccg
tgaagcgccg attcatcatc tccagagaca attccaggaa ctccctgtat 240ctgcaaaaga
acagacggag agccgaggac atggctgtgt attactgtgt gagaaa
296327296DNAArtificial SequenceSynthetic DNA IGHV3-16*02 327gaggtgcagc
tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt aacagtgaca tgaactgggc ccgcaaggct 120ccaggaaagg
ggctggagtg ggtatcgggt gttagttgga atggcagtag gacgcactat 180gtggactccg
tgaagcgccg attcatcatc tccagagaca attccaggaa ctccctgtat 240ctgcaaaaga
acagacggag agccgaggac atggctgtgt attactgtgt gagaaa
296328294DNAArtificial SequenceSynthetic DNA IGHV3/OR16-15*02
328gaggtgcagc tggtggagtc tgggggaggc ttggtccagc ctggggggtc cctgagacac
60tcctgtgcag cctctggatt caccttcagt aacagtgaca tgaactgggt cctctaggct
120ccaggaaagg ggctggagtg ggtctcgggt attagttgga atggcggtaa gacgcactat
180gtggactccg tgaagggcca atttaccatc tccagagaca attccagcaa gtccctgtat
240ctgcaaaaga acagacagag agccaaagac atggccgtgt attactgtgt gaga
294329294DNAArtificial SequenceSynthetic DNA IGHV3/OR16-16*01
329gaggtgcagc tggtggagtc tgggggaggc ttggtccagc ctggggggtc cctgagacac
60tcctgtgcag cctctggatt caccttcagt aacagtgaca tgaactgggt cctctaggct
120ccaggaaagg ggctggagtg ggtctcggat attagttgga atggcggtaa gacgcactat
180gtggactccg tgaagggcca atttaccatc tccagagaca attccagcaa gtccctgtat
240ctgcaaaaga acagacagag agccaaggac atggccgtgt attactgtgt gaga
294330296DNAArtificial SequenceSynthetic DNA IGHV3/OR16-15*01
330gaagtgcagc tggtggagtc tgggggaggc ttggtccagc ctggggggtc cctgagactc
60tcctgtgcag cctctgtatt caccttcagt aacagtgaca taaactgggt cctctaggct
120ccaggaaagg ggctggagtg ggtctcgggt attagttgga atggcggtaa gacgcactat
180gtggactccg tgaagggcca attttccatc tccagagaca attccagcaa gtccctgtat
240ctgcaaaaga acagacagag agccaaggac atggccgtgt attactgtgt gagaaa
296331296DNAArtificial SequenceSynthetic DNA IGHV3-19*01 331acagtgcagc
tggtggagtc tgggggaggc ttggtagagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt aacagtgaca tgaactgggt ccgccaggct 120ccaggaaagg
ggctggagtg ggtatcgggt gttagttgga atggcagtag gacgcactat 180gcagactctg
tgaagggccg attcatcatc tccagagaca attccaggaa cttcctgtat 240cagcaaatga
acagcctgag gcccgaggac atggctgtgt attactgtgt gagaaa
296332296DNAArtificial SequenceSynthetic DNA IGHV3-35*01 332gaggtgcagc
tggtggagtc tgggggaggc ttggtacagc ctgggggatc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt aacagtgaca tgaactgggt ccatcaggct 120ccaggaaagg
ggctggagtg ggtatcgggt gttagttgga atggcagtag gacgcactat 180gcagactctg
tgaagggccg attcatcatc tccagagaca attccaggaa caccctgtat 240ctgcaaacga
atagcctgag ggccgaggac acggctgtgt attactgtgt gagaaa
296333298DNAArtificial SequenceSynthetic DNA IGHV3-43*01 333gaagtgcagc
tggtggagtc tgggggagtc gtggtacagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt cacctttgat gattatacca tgcactgggt ccgtcaagct 120ccggggaagg
gtctggagtg ggtctctctt attagttggg atggtggtag cacatactat 180gcagactctg
tgaagggccg attcaccatc tccagagaca acagcaaaaa ctccctgtat 240ctgcaaatga
acagtctgag aactgaggac accgccttgt attactgtgc aaaagata
298334294DNAArtificial SequenceSynthetic DNA IGHV3-43*02 334gaagtgcagc
tggtggagtc tgggggaggc gtggtacagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt cacctttgat gattatgcca tgcactgggt ccgtcaagct 120ccagggaagg
gtctggagtg ggtctctctt attagtgggg atggtggtag cacatactat 180gcagactctg
tgaagggccg attcaccatc tccagagaca acagcaaaaa ctccctgtat 240ctgcaaatga
acagtctgag aactgaggac accgccttgt attactgtgc aaaa
294335298DNAArtificial SequenceSynthetic DNA IGHV3-9*01 335gaagtgcagc
tggtggagtc tgggggaggc ttggtacagc ctggcaggtc cctgagactc 60tcctgtgcag
cctctggatt cacctttgat gattatgcca tgcactgggt ccggcaagct 120ccagggaagg
gcctggagtg ggtctcaggt attagttgga atagtggtag cataggctat 180gcggactctg
tgaagggccg attcaccatc tccagagaca acgccaagaa ctccctgtat 240ctgcaaatga
acagtctgag agctgaggac acggccttgt attactgtgc aaaagata
298336296DNAArtificial SequenceSynthetic DNA IGHV3-20*01 336gaggtgcagc
tggtggagtc tgggggaggt gtggtacggc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt cacctttgat gattatggca tgagctgggt ccgccaagct 120ccagggaagg
ggctggagtg ggtctctggt attaattgga atggtggtag cacaggttat 180gcagactctg
tgaagggccg attcaccatc tccagagaca acgccaagaa ctccctgtat 240ctgcaaatga
acagtctgag agccgaggac acggccttgt atcactgtgc gagaga
296337296DNAArtificial SequenceSynthetic DNA IGHV3-74*01 337gaggtgcagc
tggtggagtc cgggggaggc ttagttcagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctactgga tgcactgggt ccgccaagct 120ccagggaagg
ggctggtgtg ggtctcacgt attaatagtg atgggagtag cacaagctac 180gcggactccg
tgaagggccg attcaccatc tccagagaca acgccaagaa cacgctgtat 240ctgcaaatga
acagtctgag agccgaggac acggctgtgt attactgtgc aagaga
296338294DNAArtificial SequenceSynthetic DNA IGHV3-74*02 338gaggtgcagc
tggtggagtc tgggggaggc ttagttcagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctactgga tgcactgggt ccgccaagct 120ccagggaagg
ggctggtgtg ggtctcacgt attaatagtg atgggagtag cacaagctac 180gcggactccg
tgaagggccg attcaccatc tccagagaca acgccaagaa cacgctgtat 240ctgcaaatga
acagtctgag agccgaggac acggctgtgt attactgtgc aaga
294339296DNAArtificial SequenceSynthetic DNA IGHV3-74*03 339gaggtgcagc
tggtggagtc cgggggaggc ttagttcagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctactgga tgcactgggt ccgccaagct 120ccagggaagg
ggctggtgtg ggtctcacgt attaatagtg atgggagtag cacaacgtac 180gcggactccg
tgaagggccg attcaccatc tccagagaca acgccaagaa cacgctgtat 240ctgcaaatga
acagtctgag agccgaggac acggctgtgt attactgtgc aagaga
296340294DNAArtificial SequenceSynthetic DNA IGHV3/OR16-13*01
340gaggtgcagc tggtggagtc tgggggaggc ttagtacagc ctggagggtc cctgagactc
60tcctgtgcag cctctggatt caccttcagt agctactgga tgcactgggt ccgccaagct
120ccagggaagg ggctggtgtg ggtctcacgt attaatagtg atgggagtag cacaagctac
180gcagactcca tgaagggcca attcaccatc tccagagaca atgctaagaa cacgctgtat
240ctgcaaatga acagtctgag agctgaggac atggctgtgt attactgtac taga
294341294DNAArtificial SequenceSynthetic DNA IGHV3/OR16-14*01
341gaggtgcagc tggaggagtc tgggggaggc ttagtacagc ctggagggtc cctgagactc
60tcctgtgcag cctctggatt caccttcagt agctactgga tgcactgggt ccgccaatct
120ccagggaagg ggctggtgtg agtctcacgt attaatagtg atgggagtag cacaagctac
180gcagactcct tgaagggcca attcaccatc tccagagaca atgctaagaa cacgctgtat
240ctgcaaatga acagtctgag agctgaggac atggctgtgt attactgtac taga
294342296DNAArtificial SequenceSynthetic DNA IGHV3-30*01 342caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatgcta tgcactgggt ccgccaggct 120ccaggcaagg
ggctagagtg ggtggcagtt atatcatatg atggaagtaa taaatactac 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agctgaggac acggctgtgt attactgtgc gagaga
296343294DNAArtificial SequenceSynthetic DNA IGHV3-30*08 343caggtgcagc
tggtggactc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cctctgcatt caccttcagt agctatgcta tgcactgggt ccgccaggct 120ccaggcaagg
ggctagagtg ggtggcagtt atatcatatg atggaagtaa taaatactac 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agctgaggac acggctgtgt attactgtgc gaga
294344296DNAArtificial SequenceSynthetic DNA IGHV3-30*17 344caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatgcta tgcactgggt ccgccaggct 120ccgggcaagg
ggctagagtg ggtggcagtt atatcatatg atggaagtaa taaatactac 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agctgaggac acggctgtgt attactgtgc gagaga
296345296DNAArtificial SequenceSynthetic DNA IGHV3-30*11 345caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cgtctggatt caccttcagt agctatgcta tgcactgggt ccgccaggct 120ccaggcaagg
ggctagagtg ggtggcagtt atatcatatg atggaagtaa taaatactac 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agctgaggac acggctgtgt attactgtgc gagaga
296346296DNAArtificial SequenceSynthetic DNA IGHV3-30*10 346caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatgcta tgcactgggt ccgccaggct 120ccaggcaagg
ggctagagtg ggtggcagtt atatcatatg atggaagtaa taaatactac 180acagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agctgaggac acggctgtgt attactgtgc gagaga
296347296DNAArtificial SequenceSynthetic DNA IGHV3-30*16 347caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatgcta tgcactgggt ccgccaggcc 120ccaggcaagg
ggctagagtg ggtggcagtt atatcatatg atggaagtaa taaatactac 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agctgaggac acggctgtgt attactgtgc gagaga
296348296DNAArtificial SequenceSynthetic DNA IGHV3-30*15 348caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatgcta tgcactgggt ccgccaggct 120ccaggcaagg
ggctagagtg ggtggcagtt atatcatatg atggaagtaa taaatactac 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
gcagcctgag agctgaggac acggctgtgt attactgtgc gagaga
296349296DNAArtificial SequenceSynthetic DNA IGHV3-30*07 349caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatgcta tgcactgggt ccgccaggct 120ccaggcaagg
ggctagagtg ggtggcagtt atatcatatg atggaagtaa taaatactac 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agccgaggac acggctgtgt attactgtgc gagaga
296350296DNAArtificial SequenceSynthetic DNA IGHV3-30*04 350caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatgcta tgcactgggt ccgccaggct 120ccaggcaagg
ggctggagtg ggtggcagtt atatcatatg atggaagtaa taaatactac 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agctgaggac acggctgtgt attactgtgc gagaga
296351296DNAArtificial SequenceSynthetic DNA IGHV3-30*09 351caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatgcta tgcactgggt ccgccaggct 120ccaggcaagg
ggctggagtg ggtggcagtt atatcatatg atggaagtaa taaatactac 180gcagactccg
tgaagggccg attcgccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agctgaggac acggctgtgt attactgtgc gagaga
296352296DNAArtificial SequenceSynthetic DNA IGHV3-30*14 352caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatgcta tgcactgggt ccgccaggct 120ccaggcaagg
ggctggagtg ggtggcagtt atatcatatg atggaagtaa taaatactac 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240cttcaaatga
acagcctgag agctgaggac acggctgtgt attactgtgc gagaga
296353294DNAArtificial SequenceSynthetic DNA IGHV3-30-3*01 353caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatgcta tgcactgggt ccgccaggct 120ccaggcaagg
ggctggagtg ggtggcagtt atatcatatg atggaagcaa taaatactac 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agctgaggac acggctgtgt attactgtgc gaga
294354296DNAArtificial SequenceSynthetic DNA IGHV3-30-3*02 354caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cgtctggatt caccttcagt agctatgcta tgcactgggt ccgccaggct 120ccaggcaagg
ggctggagtg ggtggcagtt atatcatatg atggaagcaa taaatactac 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agctgaggac acggctgtgt attactgtgc gaaaga
296355296DNAArtificial SequenceSynthetic DNA IGHV3-30*03 355caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120ccaggcaagg
ggctggagtg ggtggcagtt atatcatatg atggaagtaa taaatactat 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agctgaggac acggctgtgt attactgtgc gagaga
296356296DNAArtificial SequenceSynthetic DNA IGHV3-30*18 356caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120ccaggcaagg
ggctggagtg ggtggcagtt atatcatatg atggaagtaa taaatactat 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agctgaggac acggctgtgt attactgtgc gaaaga
296357296DNAArtificial SequenceSynthetic DNA IGHV3-30*06 357caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cgtctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120ccaggcaagg
ggctagagtg ggtggcagtt atatcatatg atggaagtaa taaatactac 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agctgaggac acggctgtgt attactgtgc gagaga
296358296DNAArtificial SequenceSynthetic DNA IGHV3-30*12 358caggtgcagc
tggtggagtc tggggggggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cgtctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120ccaggcaagg
ggctagagtg ggtggcagtt atatcatatg atggaagtaa taaatactac 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agccgaggac acggctgtgt attactgtgc gagaga
296359296DNAArtificial SequenceSynthetic DNA IGHV3-30*19 359caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cgtctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120ccaggcaagg
ggctggagtg ggtggcagtt atatcatatg atggaagtaa taaatactac 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agctgaggac acggctgtgt attactgtgc gagaga
296360296DNAArtificial SequenceSynthetic DNA IGHV3-33*05 360caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cgtctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120ccaggcaagg
ggctggagtg ggtggcagtt atatcatatg atggaagtaa taaatactat 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agccgaggac acggctgtgt attactgtgc gagaga
296361296DNAArtificial SequenceSynthetic DNA IGHV3-30*05 361caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120ccaggcaagg
ggctagagtg ggtggcagtt atatcatatg atggaagtaa taaatactac 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agctgagggc acggctgtgt attactgtgc gagaga
296362296DNAArtificial SequenceSynthetic DNA IGHV3-30*13 362caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120ccaggcaagg
ggctagagtg ggtggcagtt atatcatatg atggaagtaa taaatactac 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa caggctgtat 240ctgcaaatga
acagcctgag agctgaggac acggctgtgt attactgtgc gagaga
296363296DNAArtificial SequenceSynthetic DNA IGHV3-33*01 363caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cgtctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120ccaggcaagg
ggctggagtg ggtggcagtt atatggtatg atggaagtaa taaatactat 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agccgaggac acggctgtgt attactgtgc gagaga
296364296DNAArtificial SequenceSynthetic DNA IGHV3-33*04 364caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cgtctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120ccaggcaagg
ggctagagtg ggtggcagtt atatggtatg acggaagtaa taaatactat 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agccgaggac acggctgtgt attactgtgc gagaga
296365296DNAArtificial SequenceSynthetic DNA IGHV3-33*02 365caggtacagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cgtctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120ccaggcaagg
ggctggagtg ggtggcagtt atatggtatg atggaagtaa taaatactat 180gcagactccg
cgaagggccg attcaccatc tccagagaca attccacgaa cacgctgttt 240ctgcaaatga
acagcctgag agccgaggac acggctgtgt attactgtgc gagaga
296366296DNAArtificial SequenceSynthetic DNA IGHV3-33*03 366caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cgtctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120ccaggcaagg
ggctggagtg ggtggcagtt atatggtatg atggaagtaa taaatactat 180gcagactccg
tgaagggccg attcaccatc tccagagaca actccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agccgaggac acggctgtgt attactgtgc gaaaga
296367296DNAArtificial SequenceSynthetic DNA IGHV3-30*02 367caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctggggggtc cctgagactc 60tcctgtgcag
cgtctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120ccaggcaagg
ggctggagtg ggtggcattt atacggtatg atggaagtaa taaatactat 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agctgaggac acggctgtgt attactgtgc gaaaga
296368296DNAArtificial SequenceSynthetic DNA IGHV3-52*01 368gaggtgcagc
tggtggagtc tgggtgaggc ttggtacagc ctggagggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctcctgga tgcactgggt ctgccaggct 120ccggagaagg
ggctggagtg ggtggccgac ataaagtgtg acggaagtga gaaatactat 180gtagactctg
tgaagggccg attgaccatc tccagagaca atgccaagaa ctccctctat 240ctgcaagtga
acagcctgag agctgaggac atgaccgtgt attactgtgt gagagg
296369294DNAArtificial SequenceSynthetic DNA IGHV3-52*02 369gaggtgcagc
tggtggagtc tgggtgaggc ttggtacagc ctggagggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctcctgga tgcactgggt ctgccaggct 120ccggagaagg
ggcaggagtg ggtggccgac ataaagtgtg acggaagtga gaaatactat 180gtagactctg
tgaagggccg attgaccatc tccagagaca atgccaagaa ctccctctat 240ctgcaagtga
acagcctgag agctgaggac atgaccgtgt attactgtgt gaga
294370294DNAArtificial SequenceSynthetic DNA IGHV3-52*03 370gaggtgcagc
tggtcgagtc tgggtgaggc ttggtacagc ctggagggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctcctgga tgcactgggt ctgccaggct 120ccggagaagg
ggctggagtg ggtggccgac ataaagtgtg acggaagtga gaaatactat 180gtagactctg
tgaagggccg attgaccatc tccagagaca atgccaagaa ctccctctat 240ctgcaagtga
acagcctgag agctgaggac atgaccgtgt attactgtgt gaga
294371296DNAArtificial SequenceSynthetic DNA IGHV3-7*01 371gaggtgcagc
tggtggagtc tgggggaggc ttggtccagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt cacctttagt agctattgga tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtggccaac ataaagcaag atggaagtga gaaatactat 180gtggactctg
tgaagggccg attcaccatc tccagagaca acgccaagaa ctcactgtat 240ctgcaaatga
acagcctgag agccgaggac acggctgtgt attactgtgc gagaga
296372294DNAArtificial SequenceSynthetic DNA IGHV3-7*02 372gaggtgcagc
tggtggagtc tgggggaggc ttggtccagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt cacctttagt agctattgga tgagctgggt ccgccaggct 120ccagggaaag
ggctggagtg ggtggccaac ataaagcaag atggaagtga gaaatactat 180gtggactctg
tgaagggccg attcaccatc tccagagaca acgccaagaa ctcactgtat 240ctgcaaatga
acagcctgag agccgaggac acggctgtgt attactgtgc gaga
294373296DNAArtificial SequenceSynthetic DNA IGHV3-23*01 373gaggtgcagc
tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt cacctttagc agctatgcca tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180gcagactccg
tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agccgaggac acggccgtat attactgtgc gaaaga
296374296DNAArtificial SequenceSynthetic DNA IGHV3-23*04 374gaggtgcagc
tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt cacctttagc agctatgcca tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180gcagactccg
tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agccgaggac acggccgtat attactgtgc gaaaga
296375296DNAArtificial SequenceSynthetic DNA IGHV3-23*02 375gaggtgcagc
tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt cacctttagc agctatgcca tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180ggagactccg
tgaagggccg gttcaccatc tcaagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agccgaggac acggccgtat attactgtgc gaaaga
296376294DNAArtificial SequenceSynthetic DNA IGHV3-23*03 376gaggtgcagc
tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt cacctttagc agctatgcca tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtctcagtt atttatagcg gtggtagtag cacatactat 180gcagactccg
tgaagggccg gttcaccatc tccagagata attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agccgaggac acggccgtat attactgtgc gaaa
294377294DNAArtificial SequenceSynthetic DNA IGHV3-23*05 377gaggtgcagc
tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt cacctttagc agctatgcca tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtctcagct atttatagca gtggtagtag cacatactat 180gcagactccg
tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agccgaggac acggccgtat attactgtgc gaaa
294378293DNAArtificial SequenceSynthetic DNA IGHV3-53*01 378gaggtgcagc
tggtggagtc tggaggaggc ttgatccagc ctggggggtc cctgagactc 60tcctgtgcag
cctctgggtt caccgtcagt agcaactaca tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtctcagtt atttatagcg gtggtagcac atactacgca 180gactccgtga
agggccgatt caccatctcc agagacaatt ccaagaacac gctgtatctt 240caaatgaaca
gcctgagagc cgaggacacg gccgtgtatt actgtgcgag aga
293379291DNAArtificial SequenceSynthetic DNA IGHV3-53*02 379gaggtgcagc
tggtggagac tggaggaggc ttgatccagc ctggggggtc cctgagactc 60tcctgtgcag
cctctgggtt caccgtcagt agcaactaca tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtctcagtt atttatagcg gtggtagcac atactacgca 180gactccgtga
agggccgatt caccatctcc agagacaatt ccaagaacac gctgtatctt 240caaatgaaca
gcctgagagc cgaggacacg gccgtgtatt actgtgcgag a
291380293DNAArtificial SequenceSynthetic DNA IGHV3-66*03 380gaggtgcagc
tggtggagtc tggaggaggc ttgatccagc ctggggggtc cctgagactc 60tcctgtgcag
cctctgggtt caccgtcagt agcaactaca tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtctcagtt atttatagct gtggtagcac atactacgca 180gactccgtga
agggccgatt caccatctcc agagacaatt ccaagaacac gctgtatctt 240caaatgaaca
gcctgagagc tgaggacacg gctgtgtatt actgtgcgag aga
293381293DNAArtificial SequenceSynthetic DNA IGHV3-53*03 381gaggtgcagc
tggtggagtc tggaggaggc ttgatccagc ctggggggtc cctgagactc 60tcctgtgcag
cctctgggtt caccgtcagt agcaactaca tgagctgggt ccgccagcct 120ccagggaagg
ggctggagtg ggtctcagtt atttatagcg gtggtagcac atactacgca 180gactctgtga
agggccgatt caccatctcc agagacaatt ccaagaacac gctgtatctt 240caaatgaaca
gcctgagagc cgaggacacg gccgtgtatt actgtgctag gga
293382293DNAArtificial SequenceSynthetic DNA IGHV3-66*01 382gaggtgcagc
tggtggagtc tgggggaggc ttggtccagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccgtcagt agcaactaca tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtctcagtt atttatagcg gtggtagcac atactacgca 180gactccgtga
agggcagatt caccatctcc agagacaatt ccaagaacac gctgtatctt 240caaatgaaca
gcctgagagc cgaggacacg gctgtgtatt actgtgcgag aga
293383293DNAArtificial SequenceSynthetic DNA IGHV3-66*04 383gaggtgcagc
tggtggagtc tgggggaggc ttggtccagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccgtcagt agcaactaca tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtctcagtt atttatagcg gtggtagcac atactacgca 180gactccgtga
agggcagatt caccatctcc agagacaatt ccaagaacac gctgtatctt 240caaatgaaca
gcctgagagc cgaggacacg gctgtgtatt actgtgcgag aca
293384291DNAArtificial SequenceSynthetic DNA IGHV3-66*02 384gaggtgcagc
tggtggagtc tgggggaggc ttggtccagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccgtcagt agcaactaca tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtctcagtt atttatagcg gtggtagcac atactacgca 180gactccgtga
agggccgatt caccatctcc agagacaatt ccaagaacac gctgtatctt 240caaatgaaca
gcctgagagc tgaggacacg gctgtgtatt actgtgcgag a
291385292DNAArtificial SequenceSynthetic DNA IGHV3-38*01 385gaggtgcagc
tggtggagtc tgggggaggc ttggtacagc ctagggggtc cctgagactc 60tcctgtgcag
cctctggatt caccgtcagt agcaatgaga tgagctggat ccgccaggct 120ccagggaagg
ggctggagtg ggtctcatcc attagtggtg gtagcacata ctacgcagac 180tccaggaagg
gcagattcac catctccaga gacaattcca agaacacgct gtatcttcaa 240atgaacaacc
tgagagctga gggcacggcc gcgtattact gtgccagata ta
292386292DNAArtificial SequenceSynthetic DNA IGHV3-38*02 386gaggtgcagc
tggtggagtc tgggggaggc ttggtacagc ctagggggtc cctgagactc 60tcctgtgcag
cctctggatt caccgtcagt agcaatgaga tgagctggat ccgccaggct 120ccagggaagg
ggctggagtg ggtctcatcc attagtggtg gtagcacata ctacgcagac 180tccaggaagg
gcagattcac catctccaga gacaattcca agaacacgct gtatcttcaa 240atgaacaacc
tgagagctga gggcacggcc gtgtattact gtgccagata ta
292387288DNAArtificial SequenceSynthetic DNA IGHV3-d*01 387gaggtgcagc
tggtggagtc tcggggagtc ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccgtcagt agcaatgaga tgagctgggt ccgccaggct 120ccagggaagg
gtctggagtg ggtctcatcc attagtggtg gtagcacata ctacgcagac 180tccaggaagg
gcagattcac catctccaga gacaattcca agaacacgct gcatcttcaa 240atgaacagcc
tgagagctga ggacacggct gtgtattact gtaagaaa
288388294DNAArtificial SequenceSynthetic DNA IGHV3/OR16-12*01
388gaggtgcagc tggtagagtc tgggagaggc ttggcccagc ctggggggta cctaaaactc
60tccggtgcag cctctggatt caccgtcggt agctggtaca tgagctggat ccaccaggct
120ccagggaagg gtctggagtg ggtctcatac attagtagta gtggttgtag cacaaactac
180gcagactctg tgaagggcag attcaccatc tccacagaca actcaaagaa cacgctctac
240ctgcaaatga acagcctgag agtggaggac acggccgtgt attactgtgc aaga
294389302DNAArtificial SequenceSynthetic DNA IGHV3-15*01 389gaggtgcagc
tggtggagtc tgggggaggc ttggtaaagc ctggggggtc ccttagactc 60tcctgtgcag
cctctggatt cactttcagt aacgcctgga tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggttggccgt attaaaagca aaactgatgg tgggacaaca 180gactacgctg
cacccgtgaa aggcagattc accatctcaa gagatgattc aaaaaacacg 240ctgtatctgc
aaatgaacag cctgaaaacc gaggacacag ccgtgtatta ctgtaccaca 300ga
302390302DNAArtificial SequenceSynthetic DNA IGHV3-15*02 390gaggtgcagc
tggtggagtc tgggggagcc ttggtaaagc ctggggggtc ccttagactc 60tcctgtgcag
cctctggatt cactttcagt aacgcctgga tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggttggccgt attaaaagca aaactgatgg tgggacaaca 180gactacgctg
cacccgtgaa aggcagattc accatctcaa gagatgattc aaaaaacacg 240ctgtatctgc
aaatgaacag cctgaaaacc gaggacacag ccgtgtatta ctgtaccaca 300ga
302391302DNAArtificial SequenceSynthetic DNA IGHV3-15*04 391gaggtgcagc
tggtggagtc tgggggaggc ttggtaaagc ctggggggtc ccttagactc 60tcctgtgcag
cctctggatt cactttcagt aacgcctgga tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggttggccgt attgaaagca aaactgatgg tgggacaaca 180gactacgctg
cacccgtgaa aggcagattc accatctcaa gagatgattc aaaaaacacg 240ctgtatctgc
aaatgaacag cctgaaaacc gaggacacag ccgtgtatta ctgtaccaca 300ga
302392302DNAArtificial SequenceSynthetic DNA IGHV3-15*05 392gaggtgcagc
tggtggagtc tgggggaggc ttggtaaagc ctggggggtc ccttagactc 60tcctgtgcag
cctctggatt cactttcagt aacgcctgga tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggttggccgt attaaaagca aaactgatgg tgggacaaca 180gactacgctg
cacccgtgaa aggcagattc accatctcaa gagatgattc aaaaaacacg 240ctgtatctgc
aaatgaacag tctgaaaacc gaggacacag ccgtgtatta ctgtaccaca 300ga
302393302DNAArtificial SequenceSynthetic DNA IGHV3-15*06 393gaggtgcagc
tggtggagtc tgggggaggc ttggtaaagc ctggggggtc ccttagactc 60tcctgtgcag
cctctggatt cactttcagt aacgcctgga tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtcggccgt attaaaagca aaactgatgg tgggacaaca 180aactacgctg
cacccgtgaa aggcagattc accatctcaa gagatgattc aaaaaacacg 240ctgtatctgc
aaatgaacag cctgaaaacc gaggacacag ccgtgtatta ctgtaccaca 300ga
302394302DNAArtificial SequenceSynthetic DNA IGHV3-15*07 394gaggtgcagc
tggtggagtc tgggggaggc ttggtaaagc ctggggggtc ccttagactc 60tcctgtgcag
cctctggttt cactttcagt aacgcctgga tgaactgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtcggccgt attaaaagca aaactgatgg tgggacaaca 180gactacgctg
cacccgtgaa aggcagattc accatctcaa gagatgattc aaaaaacacg 240ctgtatctgc
aaatgaacag cctgaaaacc gaggacacag ccgtgtatta ctgtaccaca 300ga
302395302DNAArtificial SequenceSynthetic DNA IGHV3-15*03 395gaggtgcagc
tggtggagtc tgccggagcc ttggtacagc ctggggggtc ccttagactc 60tcctgtgcag
cctctggatt cacttgcagt aacgcctgga tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggttggccgt attaaaagca aagctaatgg tgggacaaca 180gactacgctg
cacctgtgaa aggcagattc accatctcaa gagttgattc aaaaaacacg 240ctgtatctgc
aaatgaacag cctgaaaacc gaggacacag ccgtgtatta ctgtaccaca 300ga
302396302DNAArtificial SequenceSynthetic DNA IGHV3-15*08 396gaggtgcagc
tggtggagtc tgcgggaggc ttggtacagc ctggggggtc ccttagactc 60tcctgtgcag
cctctggatt cacttgcagt aacgcctgga tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggttggctgt attaaaagca aagctaatgg tgggacaaca 180gactacgctg
cacctgtgaa aggcagattc accatctcaa gagatgattc aaaaaacacg 240ctgtatctgc
aaatgatcag cctgaaaacc gaggacacgg ccgtgtatta ctgtaccaca 300gg
302397302DNAArtificial SequenceSynthetic DNA IGHV3-72*01 397gaggtgcagc
tggtggagtc tgggggaggc ttggtccagc ctggagggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt gaccactaca tggactgggt ccgccaggct 120ccagggaagg
ggctggagtg ggttggccgt actagaaaca aagctaacag ttacaccaca 180gaatacgccg
cgtctgtgaa aggcagattc accatctcaa gagatgattc aaagaactca 240ctgtatctgc
aaatgaacag cctgaaaacc gaggacacgg ccgtgtatta ctgtgctaga 300ga
302398165DNAArtificial SequenceSynthetic DNA IGHV3-72*02 398accttcagtg
accactacat ggactgggtc cgccaggctc cagggaaggg gctggagtgg 60gttggccgta
ctagaaacaa agctaacagc tacaccacag aatacgccgc gtctgtgaaa 120ggcagattca
ccatctcaag agatgattca aagaactcac tgtat
165399300DNAArtificial SequenceSynthetic DNA IGHV3/OR15-7*01
399gaggtgcagc tggtggagtc tgggggaggc ttggtccagc ctgggggttc tctgagactc
60tcatgtgcag cctctggatt caccttcagt gaccactaca tgagctgggt ccgccaggct
120caagggaaag ggctagagtt ggtaggttta ataagaaaca aagctaacag ttacacgaca
180gaatatgctg cgtctgtgaa aggcagactt accatctcaa gagaggattc aaagaacacg
240atgtatctgc aaatgagcaa cctgaaaacc gaggacttgg ccgtgtatta ctgtgctaga
300400300DNAArtificial SequenceSynthetic DNA IGHV3/OR15-7*03
400gaggtgcagc tggtggagtc tgggggaggc ttggtccagc ctgggggttc tctgagactc
60tcatgtgcag cctctggatt caccttcagt gaccactaca tgagctgggt ccgccaggct
120caagggaaag ggctagagtt ggtaggttta ataagaaaca aagctaacag ttacacgaca
180gaatatgctg cgtctgtgaa aggcagactt accatctcaa gagaggattc aaagaacacg
240ctgtatctgc aaatgagcag cctgaaaacc gaggacttgg ccgtgtatta ctgtgctaga
300401300DNAArtificial SequenceSynthetic DNA IGHV3/OR15-7*02
401gaggtgcagc tgttggagtc tgggggaggc ttggtccagc ctgggggttc tctgagactc
60tcatgtgctg cctctggatt caccttcagt gaccactaca tgagctgggt ccgccaggct
120caagggaaag ggctagagtt ggtaggttta ataagaaaca aagctaacag ttacacgaca
180gaatatgctg cgtctgtgaa aggcagactt accatctcaa gagaggattc aaagaacacg
240ctgtatctgc aaatgagcag cctgaaaacc gaggacttgg ccgtgtatta ctgtgctaga
300402302DNAArtificial SequenceSynthetic DNA IGHV3-73*01 402gaggtgcagc
tggtggagtc tgggggaggc ttggtccagc ctggggggtc cctgaaactc 60tcctgtgcag
cctctgggtt caccttcagt ggctctgcta tgcactgggt ccgccaggct 120tccgggaaag
ggctggagtg ggttggccgt attagaagca aagctaacag ttacgcgaca 180gcatatgctg
cgtcggtgaa aggcaggttc accatctcca gagatgattc aaagaacacg 240gcgtatctgc
aaatgaacag cctgaaaacc gaggacacgg ccgtgtatta ctgtactaga 300ca
302403302DNAArtificial SequenceSynthetic DNA IGHV3-73*02 403gaggtgcagc
tggtggagtc cgggggaggc ttggtccagc ctggggggtc cctgaaactc 60tcctgtgcag
cctctgggtt caccttcagt ggctctgcta tgcactgggt ccgccaggct 120tccgggaaag
ggctggagtg ggttggccgt attagaagca aagctaacag ttacgcgaca 180gcatatgctg
cgtcggtgaa aggcaggttc accatctcca gagatgattc aaagaacacg 240gcgtatctgc
aaatgaacag cctgaaaacc gaggacacgg ccgtgtatta ctgtactaga 300ca
302404302DNAArtificial SequenceSynthetic DNA IGHV3-22*01 404gaggtgcatc
tggtggagtc tgggggagcc ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt tactactaca tgagcggggt ccgccaggct 120cccgggaagg
ggctggaatg ggtaggtttc attagaaaca aagctaatgg tgggacaaca 180gaatagacca
cgtctgtgaa aggcagattc acaatctcaa gagatgattc caaaagcatc 240acctatctgc
aaatgaagag cctgaaaacc gaggacacgg ccgtgtatta ctgttccaga 300ga
302405302DNAArtificial SequenceSynthetic DNA IGHV3-22*02 405gaggtgcagc
tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt tactactaca tgagcggggt ccgccaggct 120cccgggaagg
ggctggaatg ggtaggtttc attagaaaca aagctaatgg tgggacaaca 180gaatagacca
cgtctgtgaa aggcagattc acaatctcaa gagatgattc caaaagcatc 240acctatctgc
aaatgaagag cctgaaaacc gaggacacgg ccgtgtatta ctgttccaga 300ga
302406302DNAArtificial SequenceSynthetic DNA IGHV3-71*01 406gaggtgcagc
tggtggagtc cgggggaggc ttggtccagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt gactactaca tgagctgggt ccgccaggct 120cccgggaagg
ggctggagtg ggtaggtttc attagaaaca aagctaatgg tgggacaaca 180gaatagacca
cgtctgtgaa aggcagattc acaatctcaa gagatgattc caaaagcatc 240acctatctgc
aaatgaacag cctgagagcc gaggacacgg ccgtgtatta ctgtgcgaga 300ga
302407302DNAArtificial SequenceSynthetic DNA IGHV3-49*03 407gaggtgcagc
tggtggagtc tgggggaggc ttggtacagc cagggcggtc cctgagactc 60tcctgtacag
cttctggatt cacctttggt gattatgcta tgagctggtt ccgccaggct 120ccagggaagg
ggctggagtg ggtaggtttc attagaagca aagcttatgg tgggacaaca 180gaatacgccg
cgtctgtgaa aggcagattc accatctcaa gagatgattc caaaagcatc 240gcctatctgc
aaatgaacag cctgaaaacc gaggacacag ccgtgtatta ctgtactaga 300ga
302408302DNAArtificial SequenceSynthetic DNA IGHV3-49*05 408gaggtgcagc
tggtggagtc tgggggaggc ttggtaaagc cagggcggtc cctgagactc 60tcctgtacag
cttctggatt cacctttggt gattatgcta tgagctggtt ccgccaggct 120ccagggaagg
ggctggagtg ggtaggtttc attagaagca aagcttatgg tgggacaaca 180gaatacgccg
cgtctgtgaa aggcagattc accatctcaa gagatgattc caaaagcatc 240gcctatctgc
aaatgaacag cctgaaaacc gaggacacag ccgtgtatta ctgtactaga 300ga
302409302DNAArtificial SequenceSynthetic DNA |IGHV3-49*01 409gaggtgcagc
tggtggagtc tgggggaggc ttggtacagc cagggcggtc cctgagactc 60tcctgtacag
cttctggatt cacctttggt gattatgcta tgagctggtt ccgccaggct 120ccagggaagg
ggctggagtg ggtaggtttc attagaagca aagcttatgg tgggacaaca 180gaatacaccg
cgtctgtgaa aggcagattc accatctcaa gagatggttc caaaagcatc 240gcctatctgc
aaatgaacag cctgaaaacc gaggacacag ccgtgtatta ctgtactaga 300ga
302410302DNAArtificial SequenceSynthetic DNA IGHV3-49*04 410gaggtgcagc
tggtggagtc tgggggaggc ttggtacagc cagggcggtc cctgagactc 60tcctgtacag
cttctggatt cacctttggt gattatgcta tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtaggtttc attagaagca aagcttatgg tgggacaaca 180gaatacgccg
cgtctgtgaa aggcagattc accatctcaa gagatgattc caaaagcatc 240gcctatctgc
aaatgaacag cctgaaaacc gaggacacag ccgtgtatta ctgtactaga 300ga
302411302DNAArtificial SequenceSynthetic DNA IGHV3-49*02 411gaggtgcagc
tggtggagtc tgggggaggc ttggtacagc cagggccgtc cctgagactc 60tcctgtacag
cttctggatt cacctttggg tattatccta tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtaggtttc attagaagca aagcttatgg tgggacaaca 180gaatacgccg
cgtctgtgaa aggcagattc accatctcaa gagatgattc caaaagcatc 240gcctatctgc
aaatgaacag cctgaaaacc gaggacacag ccgtgtatta ctgtactaga 300ga
302412296DNAArtificial SequenceSynthetic DNA IGHV3-25*01 412gagatgcagc
tggtggagtc tgggggaggc ttgcaaaagc ctgcgtggtc cccgagactc 60tcctgtgcag
cctctcaatt caccttcagt agctactaca tgaactgtgt ccgccaggct 120ccagggaatg
ggctggagtt ggtttgacaa gttaatccta atgggggtag cacatacctc 180atagactccg
gtaaggaccg attcaatacc tccagagata acgccaagaa cacacttcat 240ctgcaaatga
acagcctgaa aaccgaggac acggccctct attagtgtac cagaga
296413296DNAArtificial SequenceSynthetic DNA IGHV3-25*02 413gagatgcagc
tggtggagtc tgggggaggc ttggcaaagc ctgcgtggtc cccgagactc 60tcctgtgcag
cctctcaatt caccttcagt agctactaca tgaactgtgt ccgccaggct 120ccagggaatg
ggctggagtt ggtttgacaa gttaatccta atgggggtag cacatacctc 180atagactccg
gtaaggaccg attcaatacc tccagagata acgccaagaa cacacttcat 240ctgcaaatga
acagcctgaa aaccgaggac acggccctct attagtgtac cagaga
296414294DNAArtificial SequenceSynthetic DNA IGHV3-25*03 414gagatgcagc
tggtggagtc tgggggaggc ttggcaaagc ctgcgtggtc cccgagactc 60tcctgtgcag
cctctcaatt caccttcagt agctactaca tgaactgtgt ccgccaggct 120ccagggaatg
ggctggagtt ggttggacaa gttaatccta atgggggtag cacatacctc 180atagactccg
gtaaggaccg attcaatacc tccagagata acgccaagaa cacacttcat 240ctgcaaatga
acagcctgaa aaccgaggac acggccctgt attagtgtac caga
294415298DNAArtificial SequenceSynthetic DNA IGHV3-63*01 415gaggtggagc
tgatagagtc catagagggc ctgagacaac ttgggaagtt cctgagactc 60tcctgtgtag
cctctggatt caccttcagt agctactgaa tgagctgggt caatgagact 120ctagggaagg
ggctggaggg agtaatagat gtaaaatatg atggaagtca gatataccat 180gcagactctg
tgaagggcag attcaccatc tccaaagaca atgctaagaa ctcaccgtat 240ctccaaacga
acagtctgag agctgaggac atgaccatgc atggctgtac ataaggtt
298416294DNAArtificial SequenceSynthetic DNA IGHV3-63*02 416gaggtggagc
tgatagagtc catagagggc ctgagacaac ttgggaagtt cctgagactc 60tcctgtgtag
cctctggatt caccttcagt agctactgaa tgagctgggt caatgagact 120ctagggaagg
ggctggaggg agtaatagat gtaaaatatg atggaagtca gatataccat 180gcagactctg
tgaagggcag attcaccatc tccaaagaca atgctaagaa ctcaccgtat 240ctgcaaacga
acagtctgag agctgaggac atgaccatgc atggctgtac ataa
294417298DNAArtificial SequenceSynthetic DNA IGHV3-32*01 417gaggtggagc
tgatagagtc catagaggac ctgagacaac ctgggaagtt cctgagactc 60tcctgtgtag
cctctagatt cgccttcagt agcttctgaa tgagccgagt tcaccagtct 120ccaggcaagg
ggctggagtg agtaatagat ataaaagatg atggaagtca gatacaccat 180gcagactctg
tgaagggcag attctccatc tccaaagaca atgctaagaa ctctctgtat 240ctgcaaatga
acactcagag agctgaggac gtggccgtgt atggctatac ataaggtc
298418296DNAArtificial SequenceSynthetic DNA IGHV3-54*01 418gaggtacagc
tggtggagtc tgaagaaaac caaagacaac ttgggggatc cctgagactc 60tcctgtgcag
actctggatt aaccttcagt agctactgaa tgagctcaga ttcccaagct 120ccagggaagg
ggctggagtg agtagtagat atatagtagg atagaagtca gctatgttat 180gcacaatctg
tgaagagcag attcaccatc tccaaagaaa atgccaagaa ctcactctgt 240ttgcaaatga
acagtctgag agcagagggc acggccgtgt attactgtat gtgagy
296419296DNAArtificial SequenceSynthetic DNA IGHV3-54*04 419gaggtacagc
tggtggagtc tgaagaaaac caaagacaac ttgggggatc cctgagactc 60tcctgtgcag
actctggatt aaccttcagt agctactgaa tgagctcaga ttcccaggct 120ccagggaagg
ggctggagtg agtagtagat atatagtagg atagaagtca gctatgttat 180gcacaatctg
tgaagagcag attcaccatc tccaaagaaa atgccaagaa ctcactctgt 240ttgcaaatga
acagtctgag agcagagggc acggccgtgt attactgtat gtgagt
296420207DNAArtificial SequenceSynthetic DNA IGHV3-54*02 420tagctactga
atgagctcag attcccaggc tccagggaag gggctggagt gagtagtaga 60tatatagtac
gatagaagtc agatatgtta tgcacaatct gtgaagagca gattcaccat 120ctccaaagaa
aatgccaaga actcactccg tttgcaaatg aacagtctga gagcagaggg 180cacggccgtg
tattactgta tgtgagg
20742131DNAArtificial SequenceSynthetic DNA >IGHJ4_1 421tgaggagacg
gtgaccaggg ttccttggcc c
3142231DNAArtificial SequenceSynthetic DNA >IGHJ4_3 422tgaggagacg
gtgaccaggg tcccttggcc c
3142331DNAArtificial SequenceSynthetic DNA >IGHJ4_2 423tgaggagacg
gtgaccaggg ttccctggcc c
3142432DNAArtificial SequenceSynthetic DNA >IGHJ3_12 424ctgaagagac
ggtgaccatt gtcccttggc cc
3242532DNAArtificial SequenceSynthetic DNA >IGHJ6_1 425ctgaggagac
ggtgaccgtg gtcccttgcc cc
3242631DNAArtificial SequenceSynthetic DNA >IGHJ6_2 426tgaggagacg
gtgaccgtgg tcccttggcc c
3142732DNAArtificial SequenceSynthetic DNA >IGHJ6_34 427ctgaggagac
ggtgaccgtg gtccctttgc cc
3242832DNAArtificial SequenceSynthetic DNA >IGHJ2_1 428ctgaggagac
agtgaccagg gtgccacggc cc
3242932DNAArtificial SequenceSynthetic DNA >IGHJ5_1 429ctgaggagac
ggtgaccagg gttccttggc cc
3243032DNAArtificial SequenceSynthetic DNA >IGHJ5_2 430ctgaggagac
ggtgaccagg gttccctggc cc
3243132DNAArtificial SequenceSynthetic DNA >IGHJ1_1 431ctgaggagac
ggtgaccagg gtgccctggc cc
3243234DNAArtificial SequenceSynthetic DNA >IGHJSEQ4_1 432tgaggagacg
gtgaccaggg ttccttggcc ccag
3443334DNAArtificial SequenceSynthetic DNA >IGHJSEQ4_3 433tgaggagacg
gtgaccaggg tcccttggcc ccag
3443434DNAArtificial SequenceSynthetic DNA >IGHJSEQ4_2 434tgaggagacg
gtgaccaggg ttccctggcc ccag
3443535DNAArtificial SequenceSynthetic DNA >IGHJSEQ3_12 435ctgaagagac
ggtgaccatt gtcccttggc cccag
3543635DNAArtificial SequenceSynthetic DNA >IGHJSEQ6_1 436ctgaggagac
ggtgaccgtg gtcccttgcc cccag
3543734DNAArtificial SequenceSynthetic DNA >IGHJSEQ6_2 437tgaggagacg
gtgaccgtgg tcccttggcc ccag
3443835DNAArtificial SequenceSynthetic DNA >IGHJSEQ6_34 438ctgaggagac
ggtgaccgtg gtccctttgc cccag
3543935DNAArtificial SequenceSynthetic DNA >IGHJSEQ2_1 439ctgaggagac
agtgaccagg gtgccacggc cccag
3544035DNAArtificial SequenceSynthetic DNA >IGHJSEQ5_1 440ctgaggagac
ggtgaccagg gttccttggc cccag
3544135DNAArtificial SequenceSynthetic DNA >IGHJSEQ5_2 441ctgaggagac
ggtgaccagg gttccctggc cccag
3544235DNAArtificial SequenceSynthetic DNA >IGHJSEQ1_1 442ctgaggagac
ggtgaccagg gtgccctggc cccag
3544336DNAArtificial SequenceSynthetic DNA >IGHV1 443tgggtgcacc
aggtccangn acaagggctt gagtgg
3644436DNAArtificial SequenceSynthetic DNA >IGHV2 444tgggtgcgac
aggctcgngn acaacgcctt gagtgg
3644536DNAArtificial SequenceSynthetic DNA >IGHV3 445tgggtgcgcc
agatgccngn gaaaggcctg gagtgg
3644636DNAArtificial SequenceSynthetic DNA >IGHV4 446tgggtccgcc
agscyccngn gaaggggctg gagtgg
3644736DNAArtificial SequenceSynthetic DNA >IGHV5 447tgggtccgcc
aggctccngn aaaggggctg gagtgg
3644836DNAArtificial SequenceSynthetic DNA >IGHV6 448tgggtctgcc
aggctccngn gaaggggcag gagtgg
3644937DNAArtificial SequenceSynthetic DNA >IGH7_3.25p 449tgtgtccgcc
aggctccagg gaatgggctg gagttgg
3745037DNAArtificial SequenceSynthetic DNA >IGH8_3.54p 450tcagattccc
aagctccagg gaaggggctg gagtgag
3745137DNAArtificial SequenceSynthetic DNA >IGH9_3.63p 451tgggtcaatg
agactctagg gaaggggctg gagggag
3745248DNAArtificial SequenceSynthetic DNA >IGHJ4*01/1-48
452actactttga ctactggggc caaggaaccc tggtcaccgt ctcctcag
4845348DNAArtificial SequenceSynthetic DNA >IGHJ4*03/1-48
453gctactttga ctactggggc caagggaccc tggtcaccgt ctcctcag
4845448DNAArtificial SequenceSynthetic DNA >IGHJ4*02/1-48
454actactttga ctactggggc cagggaaccc tggtcaccgt ctcctcag
4845550DNAArtificial SequenceSynthetic DNA >IGHJ3*01/1-50
455tgatgctttt gatgtctggg gccaagggac aatggtcacc gtctcttcag
5045650DNAArtificial SequenceSynthetic DNA >IGHJ3*02/1-50
456tgatgctttt gatatctggg gccaagggac aatggtcacc gtctcttcag
5045763DNAArtificial SequenceSynthetic DNA >IGHJ6*01/1-63
457attactacta ctactacggt atggacgtct gggggcaagg gaccacggtc accgtctcct
60cag
6345863DNAArtificial SequenceSynthetic DNA >IGHJ6*02/1-62
458attactacta ctactacggt atggacgtct ggggccaagg gaccacggtc accgtctcct
60cag
6345963DNAArtificial SequenceSynthetic DNA >IGHJ6*04/1-63
459attactacta ctactacggt atggacgtct ggggcaaagg gaccacggtc accgtctcct
60cag
6346063DNAArtificial SequenceSynthetic DNA >IGHJ6*03/1-62
460attactacta ctactactac atggacgtct ggggcaaagg gaccacggtc accgtctcct
60cag
6346153DNAArtificial SequenceSynthetic DNA >IGHJ2*01/1-53
461ctactggtac ttcgatctct ggggccgtgg caccctggtc actgtctcct cag
5346251DNAArtificial SequenceSynthetic DNA >IGHJ5*01/1-51
462acaactggtt cgactcctgg ggccaaggaa ccctggtcac cgtctcctca g
5146351DNAArtificial SequenceSynthetic DNA >IGHJ5*02/1-51
463acaactggtt cgacccctgg ggccagggaa ccctggtcac cgtctcctca g
5146452DNAArtificial SequenceSynthetic DNA >IGHJ1*01/1-52
464gctgaatact tccagcactg gggccagggc accctggtca ccgtctcctc ag
5246561DNAArtificial SequenceSynthetic DNA >IGHJ2P*01/1-61
465ctacaagtgc ttggagcact ggggcagggc agcccggaca ccgtctccct gggaacgtca
60g
6146654DNAArtificial SequenceSynthetic DNA >IGHJ1P*01/1-54
466aaaggtgctg ggggtcccct gaacccgacc cgccctgaga ccgcagccac atca
5446752DNAArtificial SequenceSynthetic DNA >IGHJ3P*01/1-52
467cttgcggttg gacttcccag ccgacagtgg tggtctggct tctgaggggt ca
5246858DNAArtificial SequenceSynthetic DNA TRBJ1-2 468aatgatacgg
cgaccaccga gatctaccta caacggttaa cctggtcccc gaaccgaa
58469284DNAArtificial SequenceSynthetic DNA TRBV2*02 469gaacctgaag
tcacccagac tcccagccat caggtcacac agatgggaca ggaagtgatc 60ttgcactgtg
tccccatctc taatcactta tacttctatt ggtacagaca aatcttgggg 120cagaaagtcg
agtttctggt ttccttttat aataatgaaa tctcagagaa gtctgaaata 180ttcgatgatc
aattctcagt tgaaaggcct gatggatcaa atttcactct gaagatccgg 240tccacaaagc
tggaggactc agccatgtac ttctgtgcca gcag
28447033DNAArtificial SequenceSynthetic DNA Jseq 1-1 470acaactgtga
gtctggtgcc ttgtccaaag aaa
3347133DNAArtificial SequenceSynthetic DNA Jseq 1-2 471acaacggtta
acctggtccc cgaaccgaag gtg
3347233DNAArtificial SequenceSynthetic DNA Jseq 1-3 472acaacagtga
gccaacttcc ctctccaaaa tat
3347333DNAArtificial SequenceSynthetic DNA Jseq 1-4 473aagacagaga
gctgggttcc actgccaaaa aac
3347433DNAArtificial SequenceSynthetic DNA Jseq 1-5 474aggatggaga
gtcgagtccc atcaccaaaa tgc
3347533DNAArtificial SequenceSynthetic DNA Jseq 1-6 475gtcacagtga
gcctggtccc gttcccaaag tgg
3347633DNAArtificial SequenceSynthetic DNA Jseq 2-1 476agcacggtga
gccgtgtccc tggcccgaag aac
3347733DNAArtificial SequenceSynthetic DNA Jseq 2-2 477agtacggtca
gcctagagcc ttctccaaaa aac
3347833DNAArtificial SequenceSynthetic DNA Jseq 2-3 478agcactgtca
gccgggtgcc tgggccaaaa tac
3347933DNAArtificial SequenceSynthetic DNA Jseq 2-4 479agcactgaga
gccgggtccc ggcgccgaag tac
3348033DNAArtificial SequenceSynthetic DNA Jseq 2-5 480agcaccagga
gccgcgtgcc tggcccgaag tac
3348133DNAArtificial SequenceSynthetic DNA Jseq 2-6 481agcacggtca
gcctgctgcc ggccccgaaa gtc
3348233DNAArtificial SequenceSynthetic DNA Jseq 2-7 482gtgaccgtga
gcctggtgcc cggcccgaag tac
3348334DNAArtificial SequenceSynthetic DNA TRBJ1-5 483nacctaggat
ggagagtcga gtcccatcac caaa
3448458DNAArtificial SequenceSynthetic DNA TRBJ1-5 484aatgatacgg
cgaccaccga gatctaccta ggatggagag tcgagtccca tcaccaaa 58
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