Abstract:
The invention relates to new members of the GAGE family referred to as GAGE-7B and GAGE-8. There are differences between these two molecules and the previously described members of the GAGE family on the genomic DNA, complementary DNA, and amino acid level.

Description:
FIELD OF THE INVENTION  
         [0001]    This invention relates to a nucleic acid molecule which codes for a tumor rejection antigen precursor. More particularly, the invention concerns genes, whose tumor rejection antigen precursor is processed, inter alia, into at least one tumor rejection antigen that is presented by MHC molecules. The genes in question are members of the GAGE family of genes.  
         BACKGROUND AND PRIOR ART  
         [0002]    The process by which the mammalian immune system recognizes and reacts to foreign or alien materials is a complex one. An important facet of the system is the T lymphocyte, or “T cell” response. This response requires that T cells recognize and interact with complexes of cell surface molecules, referred to as human leukocyte antigens (“HLA”), or major histocompatibility complexes (“MHCs”), and peptides. The peptides are derived from larger molecules which are processed by the cells which also present the HLA/MHC molecule. See in this regard Male et al.,  Advanced Immunology  (J. P. Lipincott Company, 1987), especially chapters 6-10. The interaction of T cells and HLA/peptide complexes is restricted, requiring a T cell specific for a particular combination of an HLA molecule and a peptide. If a specific T cell is not present, there is no T cell response even if its partner complex is present. Similarly, there is no response if the specific complex is absent, but the T cell is present. This mechanism is involved in the immune system&#39;s response to foreign materials, in autoimmune pathologies, and in responses to cellular abnormalities. Much work has focused on the mechanisms by which proteins are processed into the HLA binding peptides. See, in this regard, Barinaga, Science 257: 880 (1992); Fremont et al., Science 257: 919 (1992); Matsumura et al., Science 257: 927 (1992); Latron et al., Science 257: 964 (1992). Also see Engelhard, Ann. Rev. Immunol. 12: 181-207 (1994).  
           [0003]    The mechanism by which T cells recognize cellular abnormalities has also been implicated in cancer. For example, in PCT application PCT/US92/04354, filed May 22, 1992, published on Nov. 26, 1992, and incorporated by reference, a family of genes is disclosed, which are processed into peptides which, in turn, are expressed on cell surfaces, which can lead to lysis of the tumor cells by specific CTLs cytolytic T lymphocytes, or “CTLs” hereafter. The genes are said to code for “tumor rejection antigen precursors” or “TRAP” molecules, and the peptides derived therefrom are referred to as “tumor rejection antigens” or “TRAs”. See Traversari et al., Immunogenetics 35: 145 (1992); van der Bruggen et al., Science 254: 1643 (1991), for further information on this family of genes. Also, see U.S. Pat. No. 5,342,774.  
           [0004]    In U.S. Pat. No. 5,405,940, the disclosure of which is incorporated by reference, it is explained that the MAGE genes code for proteins which are processed to nonapeptides which are then presented by an HLA-A1 molecule. The reference teaches that given the known specificity of particular peptides for particular HLA molecules, one should expect a particular peptide to preferably bind to one HLA molecule. This is important, because different individuals possess different HLA phenotypes. As a result, while identification of a particular peptide as being a partner for a specific HLA molecule has diagnostic and therapeutic ramifications, these are only relevant for individuals with that particular HLA phenotype. There is a need for further work in the area, because cellular abnormalities are not restricted to one particular HLA phenotype, and targeted therapy requires some knowledge of the phenotype of the abnormal cells at issue.  
           [0005]    In U.S. Pat. No. 5,629,166 filed incorporated by reference, the fact that the MAGE-1 expression product is processed to a second TRA is disclosed. This second TRA is presented by HLA-C clone 10 molecules. The disclosure shows that a given TRAP can yield a plurality of TRAs. Also, see U.S. Pat. No.5,554,506, incorporated by reference, teaching peptides which bind to HLA-A2.  
           [0006]    U.S. Pat. Nos. 5,530,096 and 5,487,934 incorporated by reference herein teach that tyrosinase, a molecule which is produced by some normal cells (e.g., melanocytes), is processed in tumor cells to yield peptides presented by HLA-A2 molecules.  
           [0007]    In U.S. patent application Ser. No. 08/032,978, filed Mar. 18, 1993, and incorporated by reference in its entirety, a second TRA, not derived from tyrosinase is taught to be presented by HLA-A2 molecules. The TRA is derived from a TRAP, but is coded for by a non-MAGE gene. This disclosure shows that a particular HLA molecule may present TRAs derived from different sources.  
           [0008]    In U.S. Pat. No. 5,571,711, filed Jun. 17, 1993 and incorporated by reference herein, an unrelated tumor rejection antigen precursor, the so-called “BAGE” precursor, is described. The BAGE precursor is not related to the MAGE family.  
           [0009]    A further family of genes which are processed into tumor rejection antigens is taught by U.S. Pat. Nos. 5,610,013 and 5,648,226, as well as patent applications Ser. Nos. 08/531,662 and 08/602,039, filed on Sep. 21, 1995 and Feb. 15, 1996 respectively, both of which have been allowed, and U.S. patent applications Ser. Nos. 08/669,161 and 09/012,818, filed on Jun. 24, 1996 and Jan. 23, 1998, respectively. All of these applications are incorporated by reference. They reveal that there is a family of genes, the “GAGE” genes, which are related to each other. Six members of the GAGE family are described in these references.  
           [0010]    It has now been found that there are at least two further members of the GAGE family, referred to hereafter as GAGE-7 and GAGE-8. These genes, as well as other aspects of the inventions, will be described in detail in the disclosure which follows.  
         DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS 
     
    
    
     EXAMPLE 1  
       [0011]    Melanoma cell line MZ2-MEL and cell lines derived therefrom are known. See, e.g., U.S. Pat. No. 5,342,774, incorporated by reference. One subclone, i.e., MZ2-MEL 3.0 was obtained by limiting dilution, and is described in the &#39;774 patent. A subline, i.e., MZ2-ME2.43 was derived by limiting dilution of MZ2-MEL 3.0 cells which had survived mutagen treatment. See Herin, et al, Int. J. Canc. 39:390-396 (1987); Van den Eynde, et al, Int J. Canc. 44:634-640 (1980). This subline had been used as a source of cDNA from which nucleic acid molecules encoding GAGE 1-6 were isolated. See U.S. Pat. Nos. 5,610,013; 5,648,226; application Ser. Nos. 08/531,662; 08/602,039; 08/669,661; and 09/012,818 cited supra, and Van den Eynde, et al, J. Exp. Med. 182:689-698 (1995), all of which are incorporated by reference.  
         [0012]    The cDNA library from MZ2-MEL.43 was rescreened, using the same protocols as are set forth in the above referenced patent and 1995 paper. Two additional positive clones were identified. These molecules were named GAGE-7B and GAGE-8. They are discussed further, infra. The nucleotide sequences for cDNA for these molecules are set forth as SEQ ID NO: 1 (GAGE-8), and SEQ ID NOS: 2 and 3 (GAGE-7B).  
       EXAMPLE 2  
       [0013]    These experiments describe the isolation of genomic DNA molecules encoding GAGE-7B.  
         [0014]    Peripheral blood lymphocytes (PBLs) were isolated, and grown, using standard methodologies. The genomic DNA was then isolated from the PBLS, partially digested with endonuclease Sau3A1, size fractionated using NaCl density gradient centrifugation, and then ligated into GEM-11 cloning vector, which had been digested with BamHI and EcoRI.  
         [0015]    The phage library was screened, using a probe labeled with α 32 P dCTP, consisting of nucleotides 18-309 of cDNA for GAGE-1. Conditions for this Southern hybridization was standard, as described by Sambrook et al: Molecular Cloning: A Laboratory Manual (Cold Spring Harbor Press, 1989), incorporated by reference. The washing conditions were 0.2×SSC, 0.1% SDS, at 65° C.  
         [0016]    One of the positive clones was analyzed, and found to contain an insert corresponding to GAGE-7B. The sequence, set forth at SEQ ID NO: 3, contains 5 exons, including an open reading frame over exons 2 to 5, which encodes a 117 amino acid product.  
         [0017]    The fourth intron of this sequence includes two regions which show strong homology with a region found only in GAGE-1. There is a 561 base pair segment positioned in between these regions at nucleotides 7109-7659, which corresponds to a truncated, L1 retroposon which belongs to the family of long interspersed repeated elements, or “LINE”; as described by Hutchinson, et al, in Berg, et al, eds, “Mobile DNA” (Am. Soc. Microbiol. 1989), incorporated by reference. The LINE element is flanked by a perfect 13 base pair target site duplication, and contains part of the reverse transcriptase coding region, the 3′-untranslated region, and the poly-A tail of the original retroposon.  
       EXAMPLE 3  
       [0018]    A cosmid library was prepared using genomic DNA from renal cell carcinoma cell line LE9211-RCC, following the methodologies described by Lurquin, et al, Cell 58: 293-303 (1989), and screened using the Southern hybridization method set forth in example 2, using the same probe.  
         [0019]    A cosmid was identified which contained genomic DNA for GAGE-8. Its structure was the same as that of GAGE-7B, including the LINE insertion discussed supra.  
       EXAMPLE 4  
       [0020]    These experiments describe how the chromosomal location of the GAGE genes was determined. Southern blot analysis was carried out on a panel of hamster or mouse×human somatic cell hybrids, obtained from the Human Genetic Mutant Cell Repository. The DNA from these somatic cell hybrids was isolated, digested with EcoRI, and used to prepare Southern blots, in accordance with Arden, at el, Cytogenet. Cell Genet. 53:161-165 (1990), incorporated by reference. The GAGE-1 probe, labeled with α 32 P dCTP, as described supra, was used. A single, EcoRI band of 4.3 kilobases was detected, indicating that the EcoRI sites defining the fragment are conserved in all GAGE genes. The only hybridization signal came from a hybrid containing the human X chromosome. No signal came from hybrids containing human autosomes, or the Y chromosome.  
         [0021]    Experiments were than carried out to refine the localization of the GAGE locus. Somatic cell hybrids containing only a portion of the X chromosome were analyzed via Southern hybridization, as described supra, as well as by PCR.  
         [0022]    For PCR, primers corresponding to nucleotides 453-470 of GAGE-1 cDNA (sense), and nucleotides 613-630 of GAGE-1 cDNA (antisense), were used. These should amplify a 0.7 kb fragment of genomic DNA, and a fragment consisting of nucleotides 453-630 of GAGE-1 cDNA, as set forth in U.S. Pat. No. 5,610,013 at SEQ ID NO: 1. Thirty five cycles of amplification were carried out, each cycle consisting of denaturation at 94° C. (1 minute), annealing at 50° C. (1 minute), and extension at 72° C. for 1 minute. The PCR was preceded by 3 minutes of incubation at 94° C., and was followed by a soak at 72° C. for 10 minutes. Amplified products were electrophoresed on 2% agarose gels, and were visualized by ethidium bromide staining. The analysis revealed that the GAGE genes are located in chromosomal region Xp21-Xq13.  
       EXAMPLE 5  
       [0023]    A further set of experiments were carried out to find the location of the GAGE locus, using fluorescence in situ hybridization, or “FISH”. To accomplish this, PBLs were stimulated with PHA, and cultured for 72 hours. Banded chromosomes were obtained by inoculating some cultures with 5-bromodeoxyuridine, in accordance with Lemieux, et al, Cytogenet. Cell Genet 59:311-312 (1992). Cytogenetic harvests, and slide preparations were prepared using standard methods. Slides were stored at −80° C. until used.  
         [0024]    FISH hybridization to metaphase chromosomes was carried out following Pinkel, et al, Proc. Natl. Acad. Sci USA 83:2934-2938 (1986). Briefly, slides were denatured for 2 minutes in 70% formamide/2×SSC (pH 7.0), and then dehydrated in ice cold ethanol. A cosmid which contained gDNA for GAGE-7B was used as a probe. The probe (100 ng) was labeled with digoxigenin, preannealed with 100 mg of COT-1 DNA, dissolved in buffer (50% formamide, 2×SSC), denatured at 75° C. for 5 minutes, and then applied to slides. The probes were hybridized to the material on the slides, overnight at 37° C., in a humid chamber.  
         [0025]    After the incubation, the slides were washed using standard procedures, and then analyzed using standard FITC-digoxigenin detection methods, together with an amplification protocol for dual color FISH. The slides were counterstained by mounting in an antifade solution containing 1 mg/ml phenylenediamine and 0.3 mg/ml propidium iodide. Spreads were examined, and photographed. A signal was deemed to be specific only if detected on each chromatid of a single chromosome. Chromosome identification was performed via simultaneous hybridization with the satellite repeat probe, or by R-banding, using 5-bromodeoxyuridine in accordance with Lemieux, et at, supra.  
         [0026]    These experiments indicated that the GAGE locus is in the p11.2-p11.4 region of the X chromosome.  
       EXAMPLE 6  
       [0027]    These experiments were designed to determine expression of GAGE genes in various cell and tumor types. For each type of cell assayed, total RNA was extracted, using standard guanidium-isothiocyanate procedures, as taught by e.g., Davis, et al. in Basic Methods In Molecular Biology, Elsevier Science Publishing Co., N.Y. (1986), pp. 130-135. Reverse transcription was carried out on 2 ug samples of the total RNA, using 2 mM of Oligo(dT) 15  primer, in a reaction volume of 20 ul. Portions of the resulting cDNA ({fraction (1/20)} of the product), were used in the PCR amplification. In order to amplify GAGE-1, 2, and 8, the primers used were:  
                                                                         (sense, SEQ ID NO: 4)                5′-GACCAAGACG CTACGTAG-3′           and                            (antisense, SEQ ID NO: 5)                    5′-CCATCAGGAC CATCTTCA-3′              
 
         [0028]    For GAGE-3, 4, 5, 6 &amp; 7B, the primers were:  
         [0029]    5′- GACCAAGGCG CTATGTAC-3′  
         [0030]    (sense, SEQ ID NO: 6)  
         [0031]    and SEQ ID NO: 5  
         [0032]    For all amplifications, the denaturation step was 94° C. for 5 minutes, then 30 cycles of amplification (1 minute at 94° C., 2 minutes at 58° C., 2 minutes at 72° C.), then a f extension step of 72° C. for 15 minutes. The products were analyzed by agarose gel electrophoresis, with RNA integrity being checked by reverse transcription and amplification of β-actin mRNA.  
         [0033]    When these primers are used, SEQ ID NOS: 4 and 5 produce a fragment consisting of nucleotides 107-350 of SEQ ID NO: 3. SEQ ID NOS: 5 and 6 produce a fragment consisting of nucleotides 92-335 of SEQ ID NO: 1.  
         [0034]    Table 1, which follows, shows the results. The highest fraction of positive tumors were found in melanoma, esophageal and lung carcinomas. GAGE 1, 2 and 8 was found in prostate carcinomas, breast carcinomas, and sarcomas. GAGE 3, 4, 5, 6 and 7B were not found in these tumors. No expression of GAGE was found in colorectal and renal carcinoma.  
                                                                                                                                             TABLE 1                           Expression of the GAGE genes in tumors                Number of samples               ALL GAGE                    3-6                            1,2,8   +7B   None   % of samples           Expression of GAGE-1,2,8*   +   +   −   −   expressing GAGE-1,2,7            Tumor   tested   Expression of GAGE-3,4,5,6,7B   +   −   +   −   and/or GAGE-3,4,5,6,8                    Cutaneous melanoma (primaries)   79       22   1   10   46     42%       Cutaneous melanoma (metastases)   211       79   8   26   98     54%       Esophageal squamous cell carcinoma   18       7   1   1   9     50%       Esophageal adenocarcinoma   5       1   0   0   4   20       Lung squamous cell carcinoma   83       28   4   7   44   47       Lung adenocarcinoma   42       13   2   6   21   50       Head &amp; neck carcinoma   92       21   3   5   63   31       Bladder carcinoma (superficial)   35       1   0   0   34    3       Bladder carcinoma (infiltrating)   40       8   2   6   24   40       Leukemia   76       0   0   3   73    4                  
 
       EXAMPLE 7  
       [0035]    In order to determine if expression of GAGE genes could be induced by demethylation, samples of cultured tumor and normal cells were incubated for 72 hours in culture medium containing 1 uM 5-aza-2′-deoxycytidine. SEQ ID NOS: 4 and 5, supra, were used in the amplification protocol. GAGE 1, 2, and 8 were found to have been induced in sarcoma and melanoma cell lines. All GAGE genes were found to be expressed following treatment of PHA stimulated PBLs.  
         [0036]    The foregoing examples set forth the invention, which includes isolated nucleic acid molecules which encode proteins GAGE 7B and GAGE 8. These may be, e.g., those set forth at SEQ ID NO: 1, 2 or 3, as well as all nucleic acid molecules which encode the proteins encoded by theses sequences. When GAGE-7B and GAGE-8 are compared to the other members of the GAGE family, cDNA for GAGE-8 is found to be identical to cDNA for GAGE-2 but for a single nucleotide, at nucleotide 268 (“C” in GAGE-2, versus “G” in GAGE-8). This leads to a change in the amino acid at position 74 (His in GAGE-2, Asp in GAGE-8). GAGE-7B is identical to GAGE-4, but for two nucleotides at positions 268 and 548. This first difference (“G” in GAGE-4, “C” in GAGE-8), results in a change at amino acid 74 as well (Asp in GAGE-4, His in GAGE-7B).  
         [0037]    There are further differences in the organization of the genomic DNA, as explained supra. Specifically, GAGE-8 and GAGE-7B differ from GAGEs 2-6 in that they contain two 2 0 inserts in the fourth intron. These inserts are found in GAGE-1 genomic DNA; however, GAGE-8 and 7B also contain a 561 base pair insert positioned in between these two inserts, which is not found in the genomic DNA of GAGE-1.  
         [0038]    In addition to the nucleic acid molecules discussed supra, other features of the invention include expression vectors which include the nucleic acid molecules of the invention, operably linked to a promoter. Both cDNA and genomic DNA can be used, in expression vectors of various types. These, as well as the isolated nucleic acid molecules of the invention, can be used to make recombinant eukaryotic and prokaryotic cells, which contain either the isolated nucleic acid molecules or the expression vectors of the invention. The choice of which nucleic acid molecule or which expression vector to use will be up to the skilled artisan, depending upon the application of interest.  
         [0039]    The nucleic acid molecules of the invention do include segments which correspond to peptides presented by HLA-Cw6 and HLA-A29, i.e., YRPRPRRY (GAGE 1, 2 and 8), and YYWPRPRRY (GAGE 3, 4, 5, 6 and 7B). Hence, a further aspect of the invention are recombinant cells which, in addition to including molecules which encode GAGE-7B and GAGE-8, also include one or more nucleic acid molecules which encode MHC molecules, such as HLA-Cw6 and/or HLA-A29. It is to be understood that additional genes which are processed to presented antigens may be used as well the GAGE 7B and 8 genes.  
         [0040]    Also a feature of the invention are the proteins encoded by the nucleic acid molecules of the invention. As explained, supra, these proteins are similar, but not identical to other GAGE proteins. Also, part of the invention are fragments of the proteins of the invention. In particular, these fragments compare at least the first 74 amino acids encoded by the SEQ ID NO: 1, 2 or 3, and no more than the entire molecule encoded by these sequences. These proteins are set forth at SEQ ID NOS.: 6 and 7. Also a part of the invention are those peptides, derived form GAGE 7B and/or GAGE 8, which complex to MHC molecules, thereby identify a particular molecule, and also in at least some cases, facilitating the proliferation of cytolytic T cells which recognize complexes of the peptide and the MHC molecule to which it binds. One or more of these peptides can be combined in compositions, which may also include one or more adjuvants, such as GM-CSF, an interleukin, an emulsifying oil such as Vitamin E, a saponin, etc.  
         [0041]    “Minigenes” can also be produced which are nucleic acid molecules that consent of nucleotides that encode these peptides. Constructs can also be prepared, such as expression vectors, which encode one or more of these peptides.  
         [0042]    An exemplary list of such peptides, with the partner MHC molecule, follows:  
                                             GAGE 7B            Position   Sequence   HLA Molecule               43-51   EGEPATQRQ   A1        9-17   YYWPRPRRY   A24       16-24   RYVQPPEMI   A24       24-32   IGPMRPEQF   A24       11-19   WPRPRRYVQ   B7       19-27   QPPEMIGPM   B7       11-19   WPRPRRYVQ   B8       1-9   MSWRGRSTY   B3501       19-27   QPPEMIGPM   B3501       28-36   RPEQFSDEV   B3501       1-9   MSWRGRSTY   B4403       33-41   DEVEPATPE   B4403       56-64   QEGEDEGAS   B4403       108-116   EEGEKQSQC   B4403       16-24   RYVQPPEMI   B5201       19-27   QPPEMIGPM   B5201       24-32   IGPMRPEQF   B5201       28-36   RPEQFSDEV   B5201        97-105   MDPPNPEEV   B5201       19-27   QPPEMIGPM   Cw0602       28-36   RPEQFSDEV   Cw0602                  
 
         [0043]    [0043]                                             GAGE 8            Position   Sequence   HLA Molecule               16-24   YVEPPEMIG   A1       42-50   EGEPATQRQ   A1       8-16   TYRPRPRRY   A24       15-23   RYVEPPEMI   A24       23-31   IGPMRPEQF   A24       10-18   RPRPRRYVE   B7       18-26   EPPEMIGPM   B7       1-9   MSWRGRSTY   B3501       18-26   EPPEMIGPM   B3501       27-35   RPEQFSDEV   B3501       1-9   MSWRGRSTY   B4403       33-41   DEVEPATPE   B4403       56-64   QEGEDEGAS   B4403       108-116   EEGEKQSQC   B4403       18-26   EPPEMIGPM   Cw0602       27-35   RPEQFSDEV   Cw0602                    
         [0044]    Other features of the invention will be clear to the skilled artisan, and will not be set forth here.  
         [0045]    The terms and expressions which have been employed are used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, it being recognized that various modifications are possible within the scope of the invention.  
                                             GAGE-B cDNA                                1   ctgtgaggca gtgctgtgtg gttcctgccg tccggactct ttttcctcta ctgagattca                   61   tctgtgtgaa atatgagttg gcgaggaaga tcgacctatc ggcctagacc aagacgctac               121   gtagagcctc ctgaaatgat tgggcctatg cggcccgagc agttcagtga tgaagtggaa               181   ccagcaacac ctgaagaagg ggaaccagca actcaacgtc aggatcctgc agctgctcag               241   gagggagagg atgagggagc atctgcaggt caagggccga agcctgaagc tgatagccag               301   gaacagggtc acccacagac tgggtgtgag tgtgaagatg gtcctgatgg gcaggagatg               361   gacccgccaa atccagagga ggtgaaaacg cctgaagaag gtgaaaagca atcacagtgt               421   taaaagaaga cacgttgaaa tgatgcaggc tgctcctatg ttggaaattt gttcattaaa               481   attctcccaa taaagcttta cagccttctg caaagaaaaa aaaaaaaaa                  
 
         [0046]    [0046]                                             GAGE 7B cDNA                                 1   tggttcctgc cgtccggact ctttttcctc tactgagatt catctgtgtg aaatatgagt                   61   tggcgaggaa gatcgaccta ttattggcct agaccaaggc gctatgtaca gcctcctgaa               121   atgattgggc ctatgcggcc cgagcagttc agtgatgaag tggaaccagc aacacctgaa               181   gaaggggaac cagcaactca acgtcaggat cctgcagctg ctcaggaggg agaggatgag               241   ggagcatctg caggtcaagg gccgaagcct gaagctcata gccaggaaca gggtcaccca               301   cagactgggt gtgagtgtga agatggtcct gatgggcagg agatggaccc gccaaatcca               361   gaggaggtga aaacgcctga agaaggtgaa aagcaatcac agtgttaaaa gaaggcacgt               421   tgaaatgatg caggctgctc ctatgttgga aatttgttca ttaaaattct cccaataaag               481   ctttacagcc ttctgcaaag aaaaaaaaaa aaaaaaaaaa aaaaaa                    
         [0047]    [0047]                                             GAGE7B gDNA                                1   gagctcgctg cagccttgac ctcctgggct caagcgctcc tcccacctca gcctcctgag                   61   tagctgtgsg tataggtaca tgccaccatg cncagctaat ttttcgatgg tttttttgtt               121   tgttttttgt agtgatgaga ttttctgatg ttgcttaggc tggtctcgaa gtcctgagct               181   caggtgatct ggccagctca gcctcccaaa atactaggat tacaggcgtg anttggcctg               241   gtctggtttt tcttatatag gggtcttatc tatataaaga ctaaagttaa tctgtgcctt               301   tgtgcgggtg ggctaagagc atgatgactt ttatcattct attgatttaa agaaaactgt               361   ccttgactta ccagtgtgta agtccatgaa agcataattc tgttgaaagc atatattgtt               421   aatgggtgtt gggaaccgtg cactttccgc tgctgtggga gcatgtcctt ggaggtacct               481   ttcatctgtt ttctcaactc caaacatctt aggaccatgg gttgtgactg gtaggactat               541   gtatcttgct gctttcaaga cggagtatat tttcacgtgg tgtcactctg gctgtcctgt               601   ttccctaata ctgtcacttc accctctgcg attctgatgc tacaaatgat agatatcgtt               661   ttagcatttt cttacgggtc ctagcgattc tattcatttt tctttcagtc tctttctctg               721   acttgttcac attgaacaat ttccttttgg gataggttgc tatttctgtt ttcgcaggtg               781   gtttacctgt cttcccagcc agtcacagtg gtccttgtcc ccatggtggg tccggggcaa               841   gagagggccc tgggttgggg gtggggttca gttgaagatg gggtgagttt tgaggggagc               901   actacttgag tcccagaggc ataggaaaca gcagagggag gtgggattcc cttatcctca               961   atgaggatgg gcatggaggg tttggggcgt ggcgctggga acggcagccc tccccagccc               1021   acagccgcgc atgctccctg ntcccgcctc agtgcgcatg ttcactgggc gtattctgcc               1081   cggccccttc gcccacgtga agaacgccag ggagctgtga ggcagtgctg tgtggttcct               1141   gccgtccgga ctctttttcc tctactgaga ttcatctggt aggtgtgcag gccagtcatc               1201   ccgggggctg aagtgtgagt gagggtggag agggcctcgg gtgggtcagg cgggtccgtt               1261   cctggtctgt ggcctccgag ggagaagggc cacgaggtta cgtacctcct tacccttcac               1321   aggctgcgag gccaccggcg gcttcgtggt cgtgaagggg cctggacggg gaggaaggtg               1381   ggccgtggag gggaggctgt caggggctca ggtgaagacg gggtgagtgc tgttgggggg               1441   atggaagtcc cgaggtgccg ggatccccga cgacacaggg cagattccct gaatgggccc               1501   ggcgggggcg aggcgggcgg tgaagaaggg gcctggcacc tgggaaggct gcggcctggc               1561   gagcgccccc cccagcggtg tggagtgcgg agcgcccgag tgagaagcac tgcaaggtct               1621   cacctccgcc atggaaggtc cgaaaacagt gggaaggagt gggcgaggca gtgcggtcca               1681   accaaacttg ttgtgagggg gggtgaatgg ctctaggaag tgggagtgtg cccaaagcag               1741   caatcacgag aattgtgatt cactagggtt ttcgtgggga gtgcacttgt gaaactaaac               1801   ctcatcagaa atgacctctg tctgcggggc gcagtggcgc tcgcctacgt agtcccagtt               1861   actggggaca ctgaggtggg aggatccctt gagcgggagg tcgaggctgc agtgagctgt               1921   gatcacgccg ctgcactcca gcctgagcaa cacagcgata ccgcgtgtcc aaaagaaatt               1981   tagaaaaaaa tgtcctctgc cttttgccac acgccttaag atgattgctc tgccagcctg               2041   gccagcagaa gtggctttgt aggcactcag acagcgtaca cacgtatgct taactctggg               2101   acttattttg agagtatttt caaaagtaaa acggaaagtt aacatttatc catggaagtg               2161   atcgaatata gcagccctgt ggagcgcacg ttcccaatca cggttgtctg ttttcagtgt               2221   gaaatatgag ttggcgagga agatcgacct attattggcc tagaccaagg cgctatgtac               2231   agcctcctga aatgattggg cctatgcggg tgagtgctta aacgttaatt cgatgttttc               2341   tattagtaga aattaatttt tgtgatagcg tcgttgcatt agtgtggaaa tgctgataaa               2401   ggtctttcct gctcataaaa aatgaggatg gcatctcatg aaggaaacat tgattctgga               2521   tgtttgttaa tgacagattg tacacatgta ttccaacaca gagtataata gcccccaaag               2581   tcctcgtgcg tcacttttct cacagtaacc tccctgtggg tggagtaacc ttattgggca               2641   tagagcatag agttggagaa atgtctttag gcttagttag gaccagaaat agctatgtat               2701   tctgtgtata tatgtaaaat tttgtatcaa taacgaaact tattttttat ttgcacaccc               2761   acacgtattc cccagcccga gcagttcagt gatgaagtgg aaccagcaac acctgaagaa               2821   ggggaaccag caactcaacg tcaggatcct gcagctgctc aggagggaga ggatgaggga               2881   gcatctgcag gtcaaggtga gggaaaggga agaagaacgt ctgctggtgt gtgcgtgtgt               2941   gtgtgttcgt gtgtgtgtgt gcacgtgtgt gtgtgttagg cattgtcaca taggaggaag               3001   aggaggaaag aaaacaatgg aaagaatgcc tgaaattgac tggaaaagcg aggaggctat               3061   gtagtttgca gcttagctta ggcaaatccc tcactatgat aaaagttctc gactttatga               3121   atgagagaat ggaggtgcca ggattgtgtg ttatccaaga acccttgact ggtgaataca               3181   acatttgtac tgtgttctaa ggtttgtgtc ttoctatcat gtatgttgct ggaaagaagg               3241   aagtgatttt gctgaaaatg cttaaaactc aaaaggcttt actgtaaggt agcttagtac               3301   tgacccaaga atagacccag ttcagaggag caggagcagc tccaaaaacc gagtcgctga               3361   atgttggccc ccgtttcctt tgattgatat ttttatatgg tacgtttgat aaaagctgga               3421   taaatgagga tactgccata caggtagctg gtttagtgat ttttctcagc ggcctttagg               3481   aggtgattaa atccttttat ggttagaaaa goaaaaacgg aattatcctg agattaacgt               3541   gagatggaaa taatttctcc gagataaaat gttttgaaag gaagcattta tgtaacggag               3601   gtcatggatt attccaggga tgcactgtta aaagttccta gaatctgact gacaacaatg               3661   cccattaatt gctgtccgcc cactccctta ttctcagtgc ggggacagta tattttctgt               3721   gattcacaaa caatgttata tttggtgctt tgttcttcac ggggttcatt tatggaatat               3781   tacctttagg accttcggac ctaaatataa ctttatttga acaaagtgaa gtttctcttt               3841   accccgatag gtaatgggtg tcgtgactgt aagatttcca tagtcctcaa atccatccag               3201   ctaatcaatc cttcagaaac tgacattgta attgtaactg aaatcctacc cacgtggtag               3961   acttcagatt tctcagctga cacacactgc tgttggtact ctagggctga atataagcat               4021   tatacatgtc ctgtggttta tccttagatt gtcatttagg agaaaggtct aaagctgggc               4081   tgaatgccat gcactcatag tcccagctac ttgggaggcc gaggtgagag gattgcttga               4141   gtcctggagt tcaagcccag cctgggaaac acagtgagac ctcattgcta ataaataaat               4201   aaatgaataa ataaataaac acataaataa attcattaaa taaataaagt tttcatggta               4261   taggaaaaca cagatgcaaa gtttttgtgc ctagtggctg gtaatgttgc aaacgtaact               4321   ccttagtgaa ctgtaccact ttagttaaga tggtaaattt taggatatct gtatttttta               4381   ccacaattgg aaattccttt cttcctaaag ttcagtgcag ttatcatata ttcttttaaa               4441   tttttactgt atgtatcttc aagacataac attcatagaa aatttgcaca gaatagtaca               4501   atgaactcat atactgttca tctggattca ccaattgtta gtagcctttc gcttcatagg               4561   tttcacatct cttccctccg tctcttaccg tgctgcccac acactcacac acacacactc               4621   acacacacat acggatatat gtttactgtt attaatgctg aattgtctcg ataaagtttc               4681   agggattatg gtcctttacc ctatgtactt gagggtgtgt atatcgtcag aacaaagaga               4741   aagtcatttc ttggatcatc actgcacaaa gataaaaatc aggaaattta acaatgagaa               4801   aatggagtca tttaatcaca gagtgcatac tcaaattttc ccagttcccc agaaaatttc               4861   ttttttcctt ttttttttct ftgttgagac ggagtctccc tctgtgggcc aggttggagg               4921   gcagtagtgc gatctcggct cactgcaacc tacacctccc aggttctagg gattctcatg               4981   cctcagcctc ccgtgtagct gggactacag gcgccggcca ctgcggtctt gaacttctgg               5041   cctcacctgc tctgcccacc ttggcatccc aaaatgtttg gattgcaggc gtgagacccc               5101   acgcccggcc cagataattt tattgatagg atttcttttt ctgatccaga gtccagttga               5161   gaatcacacc ttgcatgtgc ctttcaggtg tttttagttt cctttaacct gtaatgtttc               5221   cttaattttt cttgtcattc acgatacgga catttttgga gaggatagac cagttggttt               5281   gcagaatatt ctgtagtttg ggctttttca tgtatttttt aaaagagttt tctcactcag               5401   gagagacggg atttgagcct tgagtcattt aatacgagaa ggacaatcag aagtagaata               5461   agagagaagt gcaaaggagg cagcaaagtt gtctgagggc agtcttcgga aaggaggagg               5521   gtaatatttc gaacaccttg ttttcctgtt ttctgctaac ggactcctga aataatgttc               5581   ctgggattct tatcaacaca tttattatta cgttagctaa agctctttat ataataatac               5641   cgagagcatg aatatcattt tcttattcat attttatgtt ttactgctta aattgatacg               5701   tattttttat ttttaagggc cgaagcctga agctcatagc caggaacagg gtcacccaca               5761   gactgggtgt gagtgtgaag atggtcctga tgggcaggag atggacccgc caaatccaga               5821   ggaggtgaaa acgcctgaag aaggtaggca atccattagg catgcacatt gtagggtgtc               5881   tgtttccaca gtatcatatt gtaactctta ctatgttttt gagacggagt ctcgctctga               5941   agaccaggct ggagtgcagt ggtgccatct cggctcactg gaaattctgt ctccagggtt               6001   caagtgattc tcctgcctga gcctctggcg gagccgggct tacaggcatg ctccgccgcg               6061   cccagctaat tgttgtattt ttagtagaga cagggtttcg ttatgttgca caggttgttc               6121   ccgaactcct gacctcaggt gatccacctg cctcgaccat tgaaattgcc gggattacag               6181   gcagagccac cgtgcccgac ccagcattat atttttaata acagagaggt aacaatactg               6241   cgtctttagt aacagagttc ttatataaag gttatttgaa acgtagttca ggccccagca               6301   cccggctgat agactgtcag atagggaaac aaagtgagtc aaagctatgt tgaattaaaa               6361   gttttgagta taaatcctta aaccagtagc tcacaatttt cagatgcttt tgtaaaggtc               6421   tgcttttaat caatacataa cacgtttgta acacccatca cttggtgtga aaaatgctga               6481   agcactcatg cgggttctaa taccagctct tacagccttg gcgagattct gagtgagtcc               6541   tttcccttct aaacctatct ttggttctta tgaaaatagt gagtttaagt cagagacttt               6601   aaaaccattt tgcattccgt ttctttcata ctctgatcct gttgcataga atgcgtggga               6661   cacagagatc atctcttcgc atggtttgtt aatcacaaat catgaaaccc tggcccgagt               6721   catctgaaaa tctctgaatt gagatttcat tgtcagtaag acagtgagcg ggccctctgc               6781   ttcatcctag tttttccgtg tggagagctg aatacgtagt ataagatctt gtgaaattgt               6841   gaattctccc tcttcttggt ttgtttgttt gtttgcgaca gagtctcagt gtgtcaccca               6901   ggctggagtg cagtgatgca atttcagctc actgcaactt ctggctccca gctaaagccg               6961   tcctcccacc tcagcctccc gagtggctgg aactacatgc acaagccacc gtgcctgact               7021   acattttttt gttttcattt ttgtagagat gaggtctcac tgtgttgccc aggcagggtt               7081   tctctggctt ttaatgaaca attgcttctt ttttttcctt ttatttattt attatacttt               7141   aagttttagg gtacatgtga cgttgtgcag gttagttaca tacgtataca tgtgccatgc               7201   tgtgcgctgc acccactatc tcatcatcta gcattaggta catctcccag tgctatccct               7261   cccccctccc cccacccgac aacagtcccc agggtgtgat attccccttc ctctgtccat               7321   gtgatctcat tgttcagttc ccacctatga gtgagaatat gcggtgtttg gttttttgtt               7381   cttgcgatag tttactgaga atgatgattt cnagtttcat ccatgtccct acaaaggaca               7441   tgaactcttc attttttagg gctgcatagt attccatagt gtatatgtgc cacattttct               7501   taatccagtc tatcgttgtt ggacatttgg gttggttcca agtctttgct atcgtgaata               7561   atgccgcaat aaacatacgt gtgcaogtgt ctttatagca gcatgattta tagtcctttg               7821   ggtatatacc cagtaatggg atggctgggt caaatggtac aattgcttct taaatctttc               7681   cccacggaaa ccttgagtga ctgaaataaa tatcaaatgg cgagagaccg tttagttcgt               7741   atcatctgtg gcatgtaggt cagtgatgct cagcatgggt gtgagtaaga tgcctgtgct               7801   atgcatgctc cctgccccac tgtoagtctt catgagcoac tatttctaat aagactgtag               7861   acacacatac gatataatca tctctaatca tatcaaatgt tacatgtaag tttcagcttt               7921   agagacatga attgataaga tttaaagttg aaagaccatg actctagtac ttcctgagta               7981   atcaactgaa gtatgcttta cacatgtgtt ttccaaattg ctgactgtta attgtaagtg               8041   cttgtgactt gaaaggaagc acatgatgtt cagggaggaa attcctttta aattctgcag               8101   gtctacgctc aaagtttatg cagaggttca attgcgtgta agacacggga tcacccatag               8161   ggttctgttt ttagtccatt taataaaacc caaactgtag tgtgctttgt atgcctttag               8221   ggtcatctga ataatctgtt gctaagtcat gttcccaatc gttgtgtttc tgttacaggt               8281   gaaaagcaat cacagtgtta aaagaaggca cgttgaaatg atgcaggctg ctcctatgtt               8341   ggaaatttgt tcattaaaat tctcccaata aagctttaca gccttctgca aagaagtctt               8401   gcgcatcttt tgtgaagttt atttctagct ttttgatgct gtgaaatatg tatcattctt               8461   tgaaatcgtg tattgtaact ctctgagctg gtatgtagag acatcgttct tttttttttt               8521   ctttctttct ttgtcctctt ttgagacgga gtcttgctct gtcgcccagg ctggagtgca               8581   gtggcgcgat ctctgctcac tgcaaccccg cctcccggat tcaagcaatt gtctgcctca               8641   gcctcccgag tagctgggat tataggcacc caccagcacg ccctggctaa gttttgtgtt               8701   tttactagag atggtttcgc atcttagccg gggtgctctt gaactcctga cctcgtgatt               8761   cacctgcctt ggcctcccaa agtgctggga ttacaggcat gcacgcctcc gcgcccggtg               8821   gagacataat tcttacatat tggttttcta tccagcggcc ttgtgaaata tgcttgtgaa               8881   ttctaaagtt tacttctagg tcgttttcag tcttcaatat acagaaacat atcatcctgg               8941   aataagagca gttttgtttc cgccattttt ttttgttttt ccttttgtac tttttttgta               9001   gagacggggt tttgccatgt ttcccgggct gttgttgnnn ttttgagtgc aagtgatgca               9061   cccacgtcac ctcccacagt gctgggatta ctggcgtggg ccaggggcca cccgtggcgg               9121   gccccgtcgt tgccattgta aagagtttta tttccttttc tgattttatg gcattgcgca               9181   gacccacccg ttacaatggt gacagtggac atccttgtct tatccctgat gagaaaccga               9241   aaaatttcaa catttcgcca tcctattcac tctccttttt ttgtagacgg actttatcag               9301   agtgagtcat tgcattctgt tccaaatttg ctgagagtat tcatttgaat atatgttgat               9361   tttcatcaaa cagtgcatct atttcgatta ccacagcgtt ttttcccatt catgggttaa               9421   tatagtgaat tcgattgata aatttgtacg tttttaggtt cgattattaa aacttgagac               9481   agcgtctcac tctgtcaccg aggctggagt gcggtggtgt tatcagagct c                    
         [0048]    [0048] 
     
       
       
         1 
         
           
             27  
           
           
             1  
             528  
             DNA  
             Homo sapiens  
             
 
           
            1 

ctgtgaggca gtgctgtgtg gttcctgccg tccggactct ttttcctcta ctgagattca     60 

tctgtgtgaa atatgagttg gcgaggaaga tcgacctatc ggcctagacc aagacgctac    120 

gtagagcctc ctgaaatgat tgggcctatg cggcccgagc agttcagtga tgaagtggaa    180 

ccagcaacac ctgaagaagg ggaaccagca actcaacgtc aggatcctgc agctgctcag    240 

gagggagagg atgagggagc atctgcaggt caagggccga agcctgaagc tgatagccag    300 

gaacagggtc acccacagac tgggtgtgag tgtgaagatg gtcctgatgg gcaggagatg    360 

gacccgccaa atccagagga ggtgaaaacg cctgaagaag gtgaaaagca atcacagtgt    420 

taaaagaaga cacgttgaaa tgatgcaggc tgctcctatg ttggaaattt gttcattaaa    480 

attctcccaa taaagcttta cagccttctg caaagaaaaa aaaaaaaa                 528 

 
           
             2  
             526  
             DNA  
             Homo sapiens  
             
 
           
            2 

tggttcctgc cgtccggact ctttttcctc tactgagatt catctgtgtg aaatatgagt     60 

tggcgaggaa gatcgaccta ttattggcct agaccaaggc gctatgtaca gcctcctgaa    120 

atgattgggc ctatgcggcc cgagcagttc agtgatgaag tggaaccagc aacacctgaa    180 

gaaggggaac cagcaactca acgtcaggat cctgcagctg ctcaggaggg agaggatgag    240 

ggagcatctg caggtcaagg gccgaagcct gaagctcata gccaggaaca gggtcaccca    300 

cagactgggt gtgagtgtga agatggtcct gatgggcagg agatggaccc gccaaatcca    360 

gaggaggtga aaacgcctga agaaggtgaa aagcaatcac agtgttaaaa gaaggcacgt    420 

tgaaatgatg caggctgctc ctatgttgga aatttgttca ttaaaattct cccaataaag    480 

ctttacagcc ttctgcaaag aaaaaaaaaa aaaaaaaaaa aaaaaa                   526 

 
           
             3  
             9531  
             DNA  
             Homo sapiens  
             
               unsure  
               92,232,1041,7412,9038-9040  
               identity of several nucleotides not known  
             
           
            3 

gagctcgctg cagccttgac ctcctgggct caagcgctcc tcccacctca gcctcctgag     60 

tagctgtgag tataggtaca tgccaccatg cncagctaat ttttcgatgg tttttttgtt    120 

tgttttttgt agtgatgaga ttttctgatg ttgcttaggc tggtctcgaa gtcctgagct    180 

caggtgatct ggccagctca gcctcccaaa atactaggat tacaggcgtg anttggcctg    240 

gtctggtttt tcttatatag gggtcttatc tatataaaga ctaaagttaa tctgtgcctt    300 

tgtgcgggtg ggctaagagc atgatgactt ttatcattct attgatttaa agaaaactgt    360 

ccttgactta ccagtgtgta agtccatgaa agcataattc tgttgaaagc atatattgtt    420 

aatgggtgtt gggaaccgtg cactttccgc tgctgtggga gcatgtcctt ggaggtacct    480 

ttcatctgtt ttctcaactc caaacatctt aggaccatgg gttgtgactg gtaggactat    540 

gtatcttgct gctttcaaga cggagtatat tttcacgtgg tgtcactctg gctgtcctgt    600 

ttccctaata ctgtcacttc accctctgcg attctgatgc tacaaatgat agatatcgtt    660 

ttagcatttt cttacgggtc ctagcgattc tattcatttt tctttcagtc tctttctctg    720 

acttgttcac attgaacaat ttccttttgg gataggttgc tatttctgtt ttcgcaggtg    780 

gtttacctgt cttcccagcc agtcacagtg gtccttgtcc ccatggtggg tccggggcaa    840 

gagagggccc tgggttgggg gtggggttca gttgaagatg gggtgagttt tgaggggagc    900 

actacttgag tcccagaggc ataggaaaca gcagagggag gtgggattcc cttatcctca    960 

atgaggatgg gcatggaggg tttggggcgt ggcgctggga acggcagccc tccccagccc   1020 

acagccgcgc atgctccctg ntcccgcctc agtgcgcatg ttcactgggc gtcttctgcc   1080 

cggccccttc gcccacgtga agaacgccag ggagctgtga ggcagtgctg tgtggttcct   1140 

gccgtccgga ctctttttcc tctactgaga ttcatctggt aggtgtgcag gccagtcatc   1200 

ccgggggctg aagtgtgagt gagggtggag agggcctcgg gtgggtcagg cgggtccgtt   1260 

cctggtctgt ggcctccgag ggagaagggc cacgaggtta cgtacctcct tacccttcac   1320 

aggctgcgag gccaccggcg gcttcgtggt cgtgaagggg cctggacggg gaggaaggtg   1380 

ggccgtggag gggaggctgt caggggctca ggtgaagacg gggtgagtgc tgttgggggg   1440 

atggaagtcc cgaggtgccg ggatccccga cgacacaggg cagattccct gaatgggccc   1500 

ggcgggggcg aggcgggcgg tgaagaaggg gcctggcacc tgggaaggct gcggcctggc   1560 

gagcgccccc cccagcggtg tggagtgcgg agcgcccgag tgagaagcac tgcaaggtct   1620 

cacctccgcc atggaaggtc cgaaaacagt gggaaggagt gggcgaggca gtgcggtcca   1680 

accaaacttg ttgtgagggg gggtgaatgg ctctaggaag tgggagtgtg cccaaagcag   1740 

caatcacgag aattgtgatt cactagggtt ttcgtgggga gtgcacttgt gaaactaaac   1800 

ctcatcagaa atgacctctg tctgcggggc gcagtggcgc tcgcctacgt agtcccagtt   1860 

actggggaca ctgaggtggg aggatccctt gagcgggagg tcgaggctgc agtgagctgt   1920 

gatcacgccg ctgcactcca gcctgagcaa cacagcgata ccgcgtgtcc aaaagaaatt   1980 

tagaaaaaaa tgtcctctgc cttttgccac acgccttaag atgattgctc tgccagcctg   2040 

gccagcagaa gtggctttgt aggcactcag acagcgtaca cacgtatgct taactctggg   2100 

acttattttg agagtatttt caaaagtaaa acggcaagtt aacatttatc catggaagtg   2160 

atcgaatata gcagccctgt ggagcgcacg ttcccaatca cggttgtctg ttttcagtgt   2220 

gaaatatgag ttggcgagga agatcgacct attattggcc tagaccaagg cgctatgtac   2280 

agcctcctga aatgattggg cctatgcggg tgagtgctta aacgttaatt cgatgttttc   2340 

tattagtaga aattaatttt tgtgatagcg tcgttgcatt agtgtggaaa tgctgataaa   2400 

ggtctttcct gctcataaaa aatgaggatg gcatctcatg aaggaaacat tgattctgga   2460 

ggattttttt ttttcctctc gtgttcttca gcttttgccc atgacttctt tctccggctt   2520 

tgtttgttaa tgacagattg tacacatgta ttccaacaca gagtataata gcccccaaag   2580 

tcctcgtgcg tcacttttct cacagtaacc tccctgtggg tggagtaacc ttattgggca   2640 

tagagcatag agttggagaa atgtctttag gcttagttag gaccagaaat agctatgtat   2700 

tctgtgtata tatgtaaaat tttgtatcaa taacgaaact tattttttat ttgcacaccc   2760 

acacgtattc cccagcccga gcagttcagt gatgaagtgg aaccagcaac acctgaagaa   2820 

ggggaaccag caactcaacg tcaggatcct gcagctgctc aggagggaga ggatgaggga   2880 

gcatctgcag gtcaaggtga gggaaaggga agaagaacgt ctgctggtgt gtgcgtgtgt   2940 

gtgtgttcgt gtgtgtgtgt gcacgtgtgt gtgtgttagg cattgtcaca taggaggaag   3000 

aggaggaaag aaaacaatgg aaagaatgcc tgaaattgac tggaaaagcg aggaggctat   3060 

gtagtttgca gcttagctta ggcaaatccc tcactatgat aaaagttctc gactttatga   3120 

atgagagaat ggaggtgcca ggattgtgtg ttatccaaga acccttgact ggtgaataca   3180 

acatttgtac tgtgttctaa ggtttgtgtc ttcctatcat gtatgttgct ggaaagaagg   3240 

aagtgatttt gctgaaaatg cttaaaactc aaaaggcttt actgtaaggt agcttagtac   3300 

tgacccaaga atagacccag ttcagaggag caggagcagc tccaaaaacc gagtcgctga   3360 

atgttggccc ccgtttcctt tgattgatat ttttatatgg tacgtttgat aaaagctgga   3420 

taaatgagga tactgccata caggtagctg gtttagtgat ttttctcagc ggcctttagg   3480 

aggtgattaa atccttttat ggttagaaaa gcaaaaacgg aattatcctg agattaacgt   3540 

gagatggaaa taatttctcc gagataaaat gttttgaaag gaagcattta tgtaacggag   3600 

gtcatggatt attccaggga tgcactgtta aaagttccta gaatctgact gacaacaatg   3660 

cccattaatt gctgtccgcc cactccctta ttctcagtgc ggggacagta tattttctgt   3720 

gattcacaaa caatgttata tttggtgctt tgttcttcac ggggttcatt tatggaatat   3780 

tacctttagg accttcggac ctaaatataa ctttatttga acaaagtgaa gtttctcttt   3840 

accccgatag gtaatgggtg tcgtgactgt aagatttcca tagtcctcaa atccatccag   3900 

ctaatcaatc cttcagaaac tgacattgta attgtaactg aaatcctacc cacgtggtag   3960 

acttcagatt tctcagctga cacacactgc tgttggtact ctagggctga atataagcat   4020 

tatacatgtc ctgtggttta tccttagatt gtcatttagg agaaaggtct aaagctgggc   4080 

tgaatgccat gcactcatag tcccagctac ttgggaggcc gaggtgagag gattgcttga   4140 

gtcctggagt tcaagcccag cctgggaaac acagtgagac ctcattgcta ataaataaat   4200 

aaatgaataa ataaataaac acataaataa attcattaaa taaataaagt tttcatggta   4260 

taggaaaaca cagatgcaaa gtttttgtgc ctagtggctg gtaatgttgc aaacgtaact   4320 

ccttagtgaa ctgtaccact ttagttaaga tggtaaattt taggatatct gtatttttta   4380 

ccacaattgg aaattccttt cttcctaaag ttcagtgcag ttatcatata ttcttttaaa   4440 

tttttactgt atgtatcttc aagacataac attcatagaa aatttgcaca gaatagtaca   4500 

atgaactcat atactgttca tctggattca ccaattgtta gtagcctttc gcttcatagg   4560 

tttcacatct cttccctccg tctcttaccg tgctgcccac acactcacac acacacactc   4620 

acacacacat acggatatat gtttactgtt attaatgctg aattgtctcg ataaagtttc   4680 

agggattatg gtcctttacc ctatgtactt gagggtgtgt atatcgtcag aacaaagaga   4740 

aagtcatttc ttggatcatc actgcacaaa gataaaaatc aggaaattta acaatgagaa   4800 

aatggagtca tttaatcaca gagtgcatac tcaaattttc ccagttcccc agaaaatttc   4860 

ttttttcctt ttttttttct ttgttgagac ggagtctccc tctgtgggcc aggttggagg   4920 

gcagtagtgc gatctcggct cactgcaacc tacacctccc aggttctagg gattctcatg   4980 

cctcagcctc ccgtgtagct gggactacag gcgccggcca ctgcggtctt gaacttctgg   5040 

cctcacctgc tctgcccacc ttggcatccc aaaatgtttg gattgcaggc gtgagacccc   5100 

acgcccggcc cagataattt tattgatagg atttcttttt ctgatccaga gtccagttga   5160 

gaatcacacc ttgcatgtgc ctttcaggtg tttttagttt cctttaacct gtaatgtttc   5220 

cttaattttt cttgtcattc acgatacgga catttttgga gaggatagac cagttggttt   5280 

gcagaatatt ctgtagtttg ggctttttca tgtatttttt aaaagagttt tctcactcag   5340 

cgtttattgg tggctactca tgccatgtaa gagtctaagc gctaggagtg taagtgctgt   5400 

gagagacggg atttgagcct tgagtcattt aatacgagaa ggacaatcag aagtagaata   5460 

agagagaagt gcaaaggagg cagcaaagtt gtctgagggc agtcttcgga aaggaggagg   5520 

gtaatatttc gaacaccttg ttttcctgtt ttctgctaac ggactcctga aataatgttc   5580 

ctgggattct tatcaacaca tttattatta cgttagctaa agctctttat ataataatac   5640 

cgagagcatg aatatcattt tcttattcat attttatgtt ttactgctta aattgatacg   5700 

tattttttat ttttaagggc cgaagcctga agctcatagc caggaacagg gtcacccaca   5760 

gactgggtgt gagtgtgaag atggtcctga tgggcaggag atggacccgc caaatccaga   5820 

ggaggtgaaa acgcctgaag aaggtaggca atccattagg catgcacatt gtagggtgtc   5880 

tgtttccaca gtatcatatt gtaactctta ctatgttttt gagacggagt ctcgctctga   5940 

agaccaggct ggagtgcagt ggtgccatct cggctcactg gaaattctgt ctccagggtt   6000 

caagtgattc tcctgcctga gcctctggcg gagccgggct tacaggcatg ctccgccgcg   6060 

cccagctaat tgttgtattt ttagtagaga cagggtttcg ttatgttgca caggttgttc   6120 

ccgaactcct gacctcaggt gatccacctg cctcgaccat tgaaattgcc gggattacag   6180 

gcagagccac cgtgcccgac ccagcattat atttttaata acagagaggt aacaatactg   6240 

cgtctttagt aacagagttc ttatataaag gttatttgaa acgtagttca ggccccagca   6300 

cccggctgat agactgtcag atagggaaac aaagtgagtc aaagctatgt tgaattaaaa   6360 

gttttgagta taaatcctta aaccagtagc tcacaatttt cagatgcttt tgtaaaggtc   6420 

tgcttttaat caatacataa cacgtttgta acacccatca cttggtgtga aaaatgctga   6480 

agcactcatg cgggttctaa taccagctct tacagccttg gcgagattct gagtgagtcc   6540 

tttcccttct aaacctatct ttggttctta tgaaaatagt gagtttaagt cagagacttt   6600 

aaaaccattt tgcattccgt ttctttcata ctctgatcct gttgcataga atgcgtggga   6660 

cacagagatc atctcttcgc atggtttgtt aatcacaaat catgaaaccc tggcccgagt   6720 

catctgaaaa tctctgaatt gagatttcat tgtcagtaag acagtgagcg ggccctctgc   6780 

ttcatcctag tttttccgtg tggagagctg aatacgtagt ataagatctt gtgaaattgt   6840 

gaattctccc tcttcttggt ttgtttgttt gtttgcgaca gagtctcagt gtgtcaccca   6900 

ggctggagtg cagtgatgca atttcagctc actgcaactt ctggctccca gctaaagccg   6960 

tcctcccacc tcagcctccc gagtggctgg aactacatgc acaagccacc gtgcctgact   7020 

acattttttt gttttcattt ttgtagagat gaggtctcac tgtgttgccc aggcagggtt   7080 

tctctggctt ttaatgaaca attgcttctt ttttttcctt ttatttattt attatacttt   7140 

aagttttagg gtacatgtga cgttgtgcag gttagttaca tacgtataca tgtgccatgc   7200 

tgtgcgctgc acccactatc tcatcatcta gcattaggta catctcccag tgctatccct   7260 

cccccctccc cccacccgac aacagtcccc agggtgtgat attccccttc ctctgtccat   7320 

gtgatctcat tgttcagttc ccacctatga gtgagaatat gcggtgtttg gttttttgtt   7380 

cttgcgatag tttactgaga atgatgattt cnagtttcat ccatgtccct acaaaggaca   7440 

tgaactcttc attttttagg gctgcatagt attccatagt gtatatgtgc cacattttct   7500 

taatccagtc tatcgttgtt ggacatttgg gttggttcca agtctttgct atcgtgaata   7560 

atgccgcaat aaacatacgt gtgcacgtgt ctttatagca gcatgattta tagtcctttg   7620 

ggtatatacc cagtaatggg atggctgggt caaatggtac aattgcttct taaatctttc   7680 

cccacggaaa ccttgagtga ctgaaataaa tatcaaatgg cgagagaccg tttagttcgt   7740 

atcatctgtg gcatgtaggt cagtgatgct cagcatgggt gtgagtaaga tgcctgtgct   7800 

atgcatgctc cctgccccac tgtcagtctt catgagccac tatttctaat aagactgtag   7860 

acacacatac gatataatca tctctaatca tatcaaatgt tacatgtaag tttcagcttt   7920 

agagacatga attgataaga tttaaagttg aaagaccatg actctagtac ttcctgagta   7980 

atcaactgaa gtatgcttta cacatgtgtt ttccaaattg ctgactgtta attgtaagtg   8040 

cttgtgactt gaaaggaagc acatgatgtt cagggaggaa attcctttta aattctgcag   8100 

gtctacgctc aaagtttatg cagaggttca attgcgtgta agacacggga tcacccatag   8160 

ggttctgttt ttagtccatt taataaaacc caaactgtag tgtgctttgt atgcctttag   8220 

ggtcatctga ataatctgtt gctaagtcat gttcccaatc gttgtgtttc tgttacaggt   8280 

gaaaagcaat cacagtgtta aaagaaggca cgttgaaatg atgcaggctg ctcctatgtt   8340 

ggaaatttgt tcattaaaat tctcccaata aagctttaca gccttctgca aagaagtctt   8400 

gcgcatcttt tgtgaagttt atttctagct ttttgatgct gtgaaatatg tatcattctt   8460 

tgaaatcgtg tattgtaact ctctgagctg gtatgtagag acatcgttct tttttttttt   8520 

ctttctttct ttgtcctctt ttgagacgga gtcttgctct gtcgcccagg ctggagtgca   8580 

gtggcgcgat ctctgctcac tgcaaccccg cctcccggat tcaagcaatt gtctgcctca   8640 

gcctcccgag tagctgggat tataggcacc caccagcacg ccctggctaa gttttgtgtt   8700 

tttactagag atggtttcgc atcttggccg gggtgctctt gaactcctga cctcgtgatt   8760 

cacctgcctt ggcctcccaa agtgctggga ttacaggcat gcacgcctcc gcgcccggtg   8820 

gagacataat tcttacatat tggttttcta tccagcggcc ttgtgaaata tgcttgtgaa   8880 

ttctaaagtt tacttctagg tcgttttcag tcttcaatat acagaaacat atcatcctgg   8940 

aataagagca gttttgtttc cgccattttt ttttgttttt ccttttgtac tttttttgta   9000 

gagacggggt tttgccatgt ttcccgggct gttgttgnnn ttttgagtgc aagtgatgca   9060 

cccacgtcac ctcccacagt gctgggatta ctggcgtggg ccaggggcca cccgtggcgg   9120 

gccccgtcgt tgccattgta aagagtttta tttccttttc tgattttatg gcattgcgca   9180 

gacccacccg ttacaatggt gacagtggac atccttgtct tatccctgat gagaaaccga   9240 

aaaatttcaa catttcgcca tcctattcac tctccttttt ttgtagacgg actttatcag   9300 

agtgagtcat tgcattctgt tccaaatttg ctgagagtat tcatttgaat atatgttgat   9360 

tttcatcaaa cagtgcatct atttcgatta ccacagcgtt ttttcccatt catgggttaa   9420 

tatagtgaat tcgattgata aatttgtacg tttttaggtt cgattattaa aacttgagac   9480 

agcgtctcac tctgtcaccg aggctggagt gcggtggtgt tatcagagct c            9531 

 
           
             4  
             18  
             DNA  
             Homo sapiens  
             
 
           
            4 

gaccaagacg  ctacgtag                                                  18 

 
           
             5  
             18  
             DNA  
             Homo sapiens  
             
 
           
            5 

ccatcaggac  catcttca                                                  18 

 
           
             6  
             18  
             DNA  
             Homo sapiens  
             
 
           
            6 

gaccaaggcg  ctatgtac                                                  18 

 
           
             7  
             117  
             PRT  
             Homo sapiens  
             
 
           
            7 

Met Ser Trp Arg Gly Arg Ser Thr Tyr Tyr Trp Pro Arg Pro Arg Arg 
 1               5                  10                  15 

Tyr Val Gln Pro Gly Pro Met Arg Pro Glu Gln Phe Ser Asp Glu Val 
            20                  25                   30 

Pro Glu Met Ile Glu Pro Ala Thr Pro Glu Glu Gly Glu Pro Ala Thr 
        35                  40                   45 

Gln Arg Gln Asp Pro Ala Ala Ala Gln Glu Gly Glu Asp Glu Gly Ala 
     50                  55                  60 

Ser Ala Gly Gln Gly Pro His Pro Gln Thr Gly Lys Pro Glu Ala His 
 65                  70                  75                  80 

Ser Gln Glu Gln Gly Cys Glu Cys Glu Asp Gly Pro Asp Gly Gln Glu 
                 85                  90                  95 

Met Asp Pro Pro Asn Pro Glu Glu Val Lys Thr Pro Glu Glu Gly Glu 
            100                 105                 110 

Lys Gln Ser Gln Cys 
        115 

 
           
             8  
             116  
             PRT  
             Homo sapiens  
             
 
           
            8 

Met Ser Trp Arg Gly Arg Ser Thr Tyr Arg Pro Arg Pro Arg Arg Tyr 
 1               5                   10                  15 

Val Glu Pro Pro Glu Met Ile Gly Pro Met Arg Pro Glu Gln Phe Ser 
             20                  25                  30 

Asp Glu Val Glu Pro Ala Thr Pro Glu Glu Gly Glu Pro Ala Thr Gln 
         35                  40                  45 

Arg Gln Asp Pro Ala Ala Ala Gln Glu Gln Glu Asp Glu Gly Ala Ser 
     50                  55                  60 

Ala Gly Gln Gly Pro Lys Pro Glu Ala Asp Ser Gln Glu Gln Gly His 
65                   70                  75                  80 

Pro Gln Thr Gly Cys Glu Cys Glu Asp Gly Pro Asp Gly Gln Glu Met 
                 85                  90                  95 

Asp Pro Pro Asn Pro Glu Glu Val Lys Thr Pro Glu Glu Lys Glu Lys 
            100                 105                 110 

Gln Ser Gln Cys 
        115 

 
           
             9  
             9  
             PRT  
             Homo sapiens  
             
 
           
            9 

Glu Gly Glu Pro Ala Thr Gln Arg Gln 
                5 

 
           
             10  
             9  
             PRT  
             Homo sapiens  
             
 
           
            10 

Tyr Tyr Trp Pro Arg Pro Arg Arg Tyr 
                5 

 
           
             11  
             9  
             PRT  
             Homo sapiens  
             
 
           
            11 

Arg Tyr Val Gln Pro Pro Glu Met Ile 
                5 

 
           
             12  
             9  
             PRT  
             Homo sapiens  
             
 
           
            12 

Ile Gly Pro Met Arg Pro Glu Gln Phe 
                5 

 
           
             13  
             9  
             PRT  
             Homo sapiens  
             
 
           
            13 

Trp Pro Arg Pro Arg Arg Tyr Val Gln 
                5 

 
           
             14  
             8  
             PRT  
             Homo sapiens  
             
 
           
            14 

Tyr Pro Pro Met Ile Gly Pro Met 
                5 

 
           
             15  
             10  
             PRT  
             Homo sapiens  
             
 
           
            15 

Met Ser Trp Arg Gly Arg Ser Asp Glu Val 
                5                   10 

 
           
             16  
             9  
             PRT  
             Homo sapiens  
             
 
           
            16 

Arg Pro Glu Gln Phe Ser Asp Glu Val 
                5 

 
           
             17  
             9  
             PRT  
             Homo sapiens  
             
 
           
            17 

Asp Glu Val Glu Pro Ala Thr Pro Glu 
                5 

 
           
             18  
             9  
             PRT  
             Homo sapiens  
             
 
           
            18 

Gln Glu Gly Glu Asp Glu Gly Ala Ser 
                5 

 
           
             19  
             9  
             PRT  
             Homo sapiens  
             
 
           
            19 

Glu Glu Gly Glu Lys Gln Ser Gln Cys 
                5 

 
           
             20  
             9  
             PRT  
             Homo sapiens  
             
 
           
            20 

Met Asp Pro Pro Asn Gln Glu Glu Val 
                5 

 
           
             21  
             9  
             PRT  
             Homo sapiens  
             
 
           
            21 

Tyr Val Glu Pro Pro Glu Met Ile Gly 
                5 

 
           
             22  
             9  
             PRT  
             Homo sapiens  
             
 
           
            22 

Thr Tyr Arg Pro Arg Pro Arg Arg Tyr 
                5 

 
           
             23  
             9  
             PRT  
             Homo sapiens  
             
 
           
            23 

Arg Tyr Val Glu Pro Pro Glu Met Ile 
                5 

 
           
             24  
             9  
             PRT  
             Homo sapiens  
             
 
           
            24 

Arg Pro Arg Pro Arg Arg Tyr Val Glu 
                5 

 
           
             25  
             9  
             PRT  
             Homo sapiens  
             
 
           
            25 

Glu Pro Pro Glu Met Ile Gly Pro Met 
                5 

 
           
             26  
             8  
             PRT  
             Homo sapiens  
             
 
           
            26 

Tyr Arg Pro Arg Pro Arg Arg Tyr 
                5 

 
           
             27  
             9  
             PRT  
             Homo sapiens  
             
 
           
            27 

Tyr Tyr Trp Pro Arg Pro Arg Arg Tyr 
                5