LOCUS Exported 12145 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Lentiviral expression of EGFR M178I. ACCESSION . VERSION . KEYWORDS pHAGE-EGFR-M178I SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12145) AUTHORS Ng PK, Li J, Jeong KJ, Shao S, Chen H, Tsang YH, Sengupta S, Wang Z, Bhavana VH, Tran R, Soewito S, Minussi DC, Moreno D, Kong K, Dogruluk T, Lu H, Gao J, Tokheim C, Zhou DC, Johnson AM, Zeng J, Ip CKM, Ju Z, Wester M, Yu S, Li Y, Vellano CP, Schultz N, Karchin R, Ding L, Lu Y, Cheung LWT, Chen K, Shaw KR, Meric-Bernstam F, Scott KL, Yi S, Sahni N, Liang H, Mills GB TITLE Systematic Functional Annotation of Somatic Mutations in Cancer. JOURNAL Cancer Cell. 2018 Mar 12;33(3):450-462.e10. doi: 10.1016/j.ccell.2018.01.021. PUBMED 29533785 REFERENCE 2 (bases 1 to 12145) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12145 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 194..311 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 359..1537 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 590..1528 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1485..1505 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" protein_bind 1557..1581 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 1584..5216 /codon_start=1 /product="precursor of human epidermal growth factor receptor" /label=EGFR /translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE DHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL QIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQ WRDIVSSDFLSNMSIDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGR CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP EGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISD GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG PEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP DNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEG CPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA" protein_bind complement(5218..5242) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 5295..5336 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" misc_feature 5357..5924 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" misc_feature 5374..5926 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(5518..5535) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5745..5764 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5938..6657 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 5938..6654 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(5983..6004) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(6244..6263) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 6591..6612 /label=EGFP-C /note="EGFP, forward primer" misc_feature 6671..7259 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(6724..6744) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(7142..7153) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 7334..7567 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" primer_bind complement(7686..7704) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 7772..7876 /gene="bla" /label=AmpR promoter CDS 7877..8737 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(8095..8114) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 8908..9496 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9397..9416 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 9650..9667 /label=L4440 /note="L4440 vector, forward primer" primer_bind 9759..9778 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" protein_bind 9907..9928 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 9943..9973 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9981..9997 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 9986..10008 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 10005..10021 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 10005..10021 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 10071..10090 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" LTR 10282..10915 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 10962..11087 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 11584..11817 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 12002..12046 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" ORIGIN 1 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca 61 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga 121 gacagatcca ttcgattagt gaacggatct cgacggtatc gccgaattca caaatggcag 181 tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa 241 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa 301 ttcaaaattt tcgggtttat tacagggaca gcagagatcc agtttggact agtcgtgagg 361 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg 421 aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga 481 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt 541 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt 601 gtgtggttcc cgcgggcctg gcctctttac gggttatggc ccttgcgtgc cttgaattac 661 ttccacctgg ctgcagtacg tgattcttga tcccgagctt cgggttggaa gtgggtggga 721 gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc 781 tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt 841 cgataagtct ctagccattt aaaatttttg atgacctgct gcgacgcttt ttttctggca 901 agatagtctt gtaaatgcgg gccaagatct gcacactggt atttcggttt ttggggccgc 961 gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc 1021 gcggccaccg agaatcggac gggggtagtc tcaagctggc cggcctgctc tggtgcctgg 1081 cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt cggcaccagt 1141 tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca gggagctcaa aatggaggac 1201 gcggcgctcg ggagagcggg cgggtgagtc acccacacaa aggaaaaggg cctttccgtc 1261 ctcagccgtc gcttcatgtg actccacgga gtaccgggcg ccgtccaggc acctcgatta 1321 gttctcgagc ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga 1381 gtttccccac actgagtggg tggagactga agttaggcca gcttggcact tgatgtaatt 1441 ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc ctcagacagt 1501 ggttcaaagt ttttttcttc catttcaggt gtcgtgaagc ggccctgcag atatcaacaa 1561 gtttgtacaa aaaagcaggc accatgcgac cctccgggac ggccggggca gcgctcctgg 1621 cgctgctggc tgcgctctgc ccggcgagtc gggctctgga ggaaaagaaa gtttgccaag 1681 gcacgagtaa caagctcacg cagttgggca cttttgaaga tcattttctc agcctccaga 1741 ggatgttcaa taactgtgag gtggtccttg ggaatttgga aattacctat gtgcagagga 1801 attatgatct ttccttctta aagaccatcc aggaggtggc tggttatgtc ctcattgccc 1861 tcaacacagt ggagcgaatt cctttggaaa acctgcagat catcagagga aatatgtact 1921 acgaaaattc ctatgcctta gcagtcttat ctaactatga tgcaaataaa accggactga 1981 aggagctgcc catgagaaat ttacaggaaa tcctgcatgg cgccgtgcgg ttcagcaaca 2041 accctgccct gtgcaacgtg gagagcatcc agtggcggga catagtcagc agtgactttc 2101 tcagcaacat gtcgattgac ttccagaacc acctgggcag ctgccaaaag tgtgatccaa 2161 gctgtcccaa tgggagctgc tggggtgcag gagaggagaa ctgccagaaa ctgaccaaaa 2221 tcatctgtgc ccagcagtgc tccgggcgct gccgtggcaa gtcccccagt gactgctgcc 2281 acaaccagtg tgctgcaggc tgcacaggcc cccgggagag cgactgcctg gtctgccgca 2341 aattccgaga cgaagccacg tgcaaggaca cctgcccccc actcatgctc tacaacccca 2401 ccacgtacca gatggatgtg aaccccgagg gcaaatacag ctttggtgcc acctgcgtga 2461 agaagtgtcc ccgtaattat gtggtgacag atcacggctc gtgcgtccga gcctgtgggg 2521 ccgacagcta tgagatggag gaagacggcg tccgcaagtg taagaagtgc gaagggcctt 2581 gccgcaaagt gtgtaacgga ataggtattg gtgaatttaa agactcactc tccataaatg 2641 ctacgaatat taaacacttc aaaaactgca cctccatcag tggcgatctc cacatcctgc 2701 cggtggcatt taggggtgac tccttcacac atactcctcc tctggatcca caggaactgg 2761 atattctgaa aaccgtaaag gaaatcacag ggtttttgct gattcaggct tggcctgaaa 2821 acaggacgga cctccatgcc tttgagaacc tagaaatcat acgcggcagg accaagcaac 2881 atggtcagtt ttctcttgca gtcgtcagcc tgaacataac atccttggga ttacgctccc 2941 tcaaggagat aagtgatgga gatgtgataa tttcaggaaa caaaaatttg tgctatgcaa 3001 atacaataaa ctggaaaaaa ctgtttggga cctccggtca gaaaaccaaa attataagca 3061 acagaggtga aaacagctgc aaggccacag gccaggtctg ccatgccttg tgctcccccg 3121 agggctgctg gggcccggag cccagggact gcgtctcttg ccggaatgtc agccgaggca 3181 gggaatgcgt ggacaagtgc aaccttctgg agggtgagcc aagggagttt gtggagaact 3241 ctgagtgcat acagtgccac ccagagtgcc tgcctcaggc catgaacatc acctgcacag 3301 gacggggacc agacaactgt atccagtgtg cccactacat tgacggcccc cactgcgtca 3361 agacctgccc ggcaggagtc atgggagaaa acaacaccct ggtctggaag tacgcagacg 3421 ccggccatgt gtgccacctg tgccatccaa actgcaccta cggatgcact gggccaggtc 3481 ttgaaggctg tccaacgaat gggcctaaga tcccgtccat cgccactggg atggtggggg 3541 ccctcctctt gctgctggtg gtggccctgg ggatcggcct cttcatgcga aggcgccaca 3601 tcgttcggaa gcgcacgctg cggaggctgc tgcaggagag ggagcttgtg gagcctctta 3661 cacccagtgg agaagctccc aaccaagctc tcttgaggat cttgaaggaa actgaattca 3721 aaaagatcaa agtgctgggc tccggtgcgt tcggcacggt gtataaggga ctctggatcc 3781 cagaaggtga gaaagttaaa attcccgtcg ctatcaagga attaagagaa gcaacatctc 3841 cgaaagccaa caaggaaatc ctcgatgaag cctacgtgat ggccagcgtg gacaaccccc 3901 acgtgtgccg cctgctgggc atctgcctca cctccaccgt gcagctcatc acgcagctca 3961 tgcccttcgg ctgcctcctg gactatgtcc gggaacacaa agacaatatt ggctcccagt 4021 acctgctcaa ctggtgtgtg cagatcgcaa agggcatgaa ctacttggag gaccgtcgct 4081 tggtgcaccg cgacctggca gccaggaacg tactggtgaa aacaccgcag catgtcaaga 4141 tcacagattt tgggctggcc aaactgctgg gtgcggaaga gaaagaatac catgcagaag 4201 gaggcaaagt gcctatcaag tggatggcat tggaatcaat tttacacaga atctataccc 4261 accagagtga tgtctggagc tacggggtga ccgtttggga gttgatgacc tttggatcca 4321 agccatatga cggaatccct gccagcgaga tctcctccat cctggagaaa ggagaacgcc 4381 tccctcagcc acccatatgt accatcgatg tctacatgat catggtcaag tgctggatga 4441 tagacgcaga tagtcgccca aagttccgtg agttgatcat cgaattctcc aaaatggccc 4501 gagaccccca gcgctacctt gtcattcagg gggatgaaag aatgcatttg ccaagtccta 4561 cagactccaa cttctaccgt gccctgatgg atgaagaaga catggacgac gtggtggatg 4621 ccgacgagta cctcatccca cagcagggct tcttcagcag cccctccacg tcacggactc 4681 ccctcctgag ctctctgagt gcaaccagca acaattccac cgtggcttgc attgatagaa 4741 atgggctgca aagctgtccc atcaaggaag acagcttctt gcagcgatac agctcagacc 4801 ccacaggcgc cttgactgag gacagcatag acgacacctt cctcccagtg cctgaataca 4861 taaaccagtc cgttcccaaa aggcccgctg gctctgtgca gaatcctgtc tatcacaatc 4921 agcctctgaa ccccgcgccc agcagagacc cacactacca ggacccccac agcactgcag 4981 tgggcaaccc cgagtatctc aacactgtcc agcccacctg tgtcaacagc acattcgaca 5041 gccctgccca ctgggcccag aaaggcagcc accaaattag cctggacaac cctgactacc 5101 agcaggactt ctttcccaag gaagccaagc caaatggcat ctttaagggc tccacagctg 5161 aaaatgcaga atacctaagg gtcgcgccac aaagcagtga atttattgga gcatagaacc 5221 cagctttctt gtacaaagtg gttgatatcc agcacagtgg cggccgctcg agtctagagg 5281 gcccgcggtt cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta 5341 ccggttagta atgagatccc tccccccccc ctaacgttac tggccgaagc cgcttggaat 5401 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg 5461 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc 5521 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt 5581 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg 5641 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac 5701 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg 5761 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg 5821 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc 5881 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacacatatg 5941 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc 6001 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc 6061 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc 6121 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag 6181 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc 6241 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg 6301 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag 6361 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc 6421 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac 6481 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac 6541 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg 6601 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaatc 6661 gatagatcct aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa 6721 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat 6781 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta 6841 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc 6901 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt 6961 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg 7021 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc 7081 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc 7141 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct 7201 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc 7261 tgagatcctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa 7321 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct 7381 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg 7441 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt 7501 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc 7561 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga 7621 atatcagaga gtgagaggcc cgggttaatt aaggaaaggg ctagatcatt cttgaagacg 7681 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta 7741 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 7801 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 7861 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc 7921 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga 7981 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct 8041 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg 8101 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta 8161 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat 8221 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt 8281 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga 8341 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga 8401 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga 8461 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc 8521 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc 8581 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg 8641 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat 8701 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata 8761 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct 8821 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 8881 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 8941 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 9001 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct 9061 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 9121 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 9181 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 9241 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 9301 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 9361 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 9421 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 9481 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 9541 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac 9601 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt 9661 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat 9721 tcattaatgc agcaagctca tggctgacta atttttttta tttatgcaga ggccgaggcc 9781 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt 9841 tgcaaaaagc tccccgtggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa 9901 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc 9961 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga 10021 catgattacg aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 10081 caaactcatc aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa 10141 tcatcccaaa cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt 10201 actctaaacc tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat 10261 atgtactaca aacttagtag ttggaagggc taattcactc ccaaagaaga caagatatcc 10321 ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag 10381 ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc 10441 cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc 10501 tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag 10561 catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag 10621 cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg 10681 gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct 10741 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 10801 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 10861 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc 10921 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc 10981 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa 11041 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg 11101 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata 11161 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc 11221 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga 11281 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc 11341 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca 11401 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga 11461 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt 11521 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg 11581 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc 11641 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa 11701 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa 11761 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg 11821 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg 11881 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga 11941 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga 12001 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa 12061 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg 12121 tttaagaata gtttttgctg tactt //