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:
This application is a divisional application of U.S. Ser. No. 09/163,748 filed Sep. 30, 1998 (now U.S. Pat. No. 6,509,172). 
    
    
     FIELD OF THE INVENTION 
     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 
     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). 
     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. 
     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. 
     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. 
     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. 
     In U.S. patent application Ser. No. 08/032,978, filed Mar. 18, 1993 (now U.S. Pat. No. 5,620,886), 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-MA GB gene. This disclosure shows that a particular HLA molecule may present TRAs derived from different sources. 
     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. 
     A futher family of genes which are processed into tumor rejection antigens is taught by U.S. Pat. Nos. 6,610,013 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, now U.S. Pat. Nos. 5,858,689 and 6,069,001 respectively, and U.S. patent applications Ser. No. 08/669,161 (now U.S. Pat. No. 6,013,481) and Ser. No. 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. 
     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 
     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. 
     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 
     These experiments describe the isolation of genomic DNA molecules encoding GAGE-7B. 
     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. 
     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. 
     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. 
     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 
     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. 
     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 
     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. 
     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. 
     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 
     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. 
     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. 
     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. 
     These experiments indicated that the GAGE locus is in the p11.2-p11.4 region of the X chromosome. 
     Example 6  
     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., New York (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 ( 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′            
For GAGE-3, 4, 5, 6; &amp; 7B, the primers were:
     5′- GACCAAGGCG CTATGTAC-3′   (sense, SEQ ID NO: 6) and SEQ ID NO: 5
 
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 final 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.
   
     When these primers are used, SEQ ID NOS: 4 and 5 produce a fragment consisting of nucleotides 107-350 of SEQ ID NO: 1. SEQ ID NOS: 5 and 6 produce a fragment consisting of nucleotides 92-335 of SEQ ID NO: 2. 
     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 
     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. 
     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). 
     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 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. 
     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. 
     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. 
     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.: 7 and 8. 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 Vitainin E, a saponin, etc. 
     “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. 
     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 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
             
           
               
                   
               
               
                 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 
               
               
                   
               
             
          
         
       
     
     Other features of the invention will be clear to the skilled artisan, and will not be set forth here. 
     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-8 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 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
             
           
               
                   
               
               
                 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 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
             
           
               
                   
               
               
                 GAGE7B gDNA 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 1 
                 gagctcgctg cagccttgac ctcctgggct caagcgctcc tcccacctca gcctcctgag 
               
               
                   
               
               
                 61 
                 tagctgtgag 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 
               
               
                   
               
               
                 2281 
                 agcctcctga aatgattggg cctatgcggg tgagtgctta aacgttaatt cgatgttttc 
               
               
                   
               
               
                 2341 
                 tattagtaga aattaatttt tgtgatagcg tcgttgcatt agtgtggaaa tgctgataaa 
               
               
                   
               
               
                 2401 
                 ggtctttcct gctcataaaa aatgaggatg gcatctcatg aaggaaacat tgattctgga 
               
               
                   
               
               
                 2461 
                 ggattttttt ttttcctctc gtgttcttca gcttttgccc atgacttctt tctccggctt 
               
               
                   
               
               
                 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 gcaaaaacgg 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 
               
               
                   
               
               
                 3901 
                 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 
               
               
                   
               
               
                 5341 
                 cgtttattgg tggctactca tgccatgtaa gagtctaagc gctaggagtg taagtgctgt 
               
               
                   
               
               
                 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 
               
               
                   
               
               
                 7621 
                 ggtatatacc cagtaatggg atggctgggt caaatggtac aattgcttct taaatctttc 
               
               
                   
               
               
                 7681 
                 cccacggaaa ccttgagtga ctgaaataaa tatcaaatgg cgagagaccg tttagttcgt 
               
               
                   
               
               
                 7741 
                 atcatctgtg gcatgtaggt cagtgatgct cagcatgggt gtgagtaaga tgcctgtgct 
               
               
                   
               
               
                 7801 
                 atgcatgctc cctgccccac tgtcagtctt catgagccac 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 atcttggccg 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