LOCUS Exported 11688 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expressing base editor A3A-BE3 in yeast cells. ACCESSION . VERSION . KEYWORDS pJT166_GalL_A3A-BE3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11688) AUTHORS Tan J, Zhang F, Karcher D, Bock R TITLE Expanding the genome-targeting scope and the site selectivity of high-precision base editors. JOURNAL Nat Commun. 2020 Jan 31;11(1):629. doi: 10.1038/s41467-020-14465-z. PUBMED 32005820 REFERENCE 2 (bases 1 to 11688) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11688 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 1..589 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 490..509 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 743..760 /label=L4440 /note="L4440 vector, forward primer" protein_bind 877..898 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 913..943 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 951..967 /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 956..978 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 975..991 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 975..991 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 1010..1030 /label=T3 /note="T3 promoter, forward primer" promoter 1012..1030 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind 1439..1462 /label=GAL1 /note="S. cerevisiae GAL1 promoter, forward primer" primer_bind 1483..1499 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind 1483..1499 /label=pBluescriptSK /note="For pBluescript vector" CDS 1513..2109 /codon_start=1 /gene="human APOBEC3A" /gene_synonym="A3A" /product="human cytidine deaminase (C to U editing enzyme) that efficiently edits methylated cytosines (Wang et al., 2018)" /label=APOBEC3A /note="also known as hA3A" /translation="MEASPASGPRHLMDPHIFTSNFNNGIGRHKTYLCYEVERLDNGTS VKMDQHRGFLHNQAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTWFISWSPCFSW GCAGEVRAFLQENTHVRLRIFAARIYDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWD TFVDHQGCPFQPWDGLDEHSQALSGRLRAILQNQGN" CDS 2158..6261 /codon_start=1 /product="nickase mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9(D10A) /note="generates RNA-guided single strand nicks in DNA due to the D10A mutation in the RuvC catalytic domain" /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 6274..6522 /codon_start=1 /product="uracil-DNA glycosylase inhibitor from a Bacillus subtilis bacteriophage (Mol et al., 1995)" /label=UGI /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML" CDS 6535..6555 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" terminator 6571..6818 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(6587..6605) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" primer_bind complement(6836..6855) /label=T7 /note="T7 promoter, forward primer" promoter complement(6837..6855) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(6865..6882) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(6865..6881) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6874..6896) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 7022..7477 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7109..7128) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7319..7340 /label=F1ori-F /note="F1 origin, forward primer" promoter 7778..8185 /gene="S. cerevisiae LEU2" /label=LEU2 promoter CDS 8186..9280 /codon_start=1 /gene="S. cerevisiae LEU2" /product="3-isopropylmalate dehydrogenase, required for leucine biosynthesis" /label=LEU2 /note="yeast auxotrophic marker" /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG DLGGSNSTTEVGDAVAEEVKKILA" primer_bind complement(9774..9793) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 9893..9915 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(9953..9971) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" misc_feature 10013..10516 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" promoter 10553..10657 /gene="bla" /label=AmpR promoter CDS 10658..11518 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(10876..10895) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg 721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat 781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt 841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta 901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg 961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc 1021 tcactaaagg gaacaaaagc tggagctcgt gacagccctc cgaaggaaga ctttcctccg 1081 tgcgtcctcg tcttcaccgg tcgcgttcct gaaacgcaga tgtgcctcgc gccgcactgc 1141 tccgaacaat aaagattcta caatactagc ttttatggtt atgaagagga aaaattggca 1201 gtaacctggc cccacaaacc ttcaaatgaa cgaatcaaat taacaaccat aggatgataa 1261 tgcgattagt tttttagcct tatttctggg gtaattaatc agcgaagcga tgatttttga 1321 tctattaacg gatatataaa tgcaaaaact gcataaccac tttaactaat actttcaaca 1381 ttttcggttt gtattacttc ttattcaaat gtaataaaag tatcaacaaa aaattgttaa 1441 tatacctcta tactttaacg tcaaggagaa aaaaccccgg attctagaac tagtggatcc 1501 cccgggaaaa aaatggaagc cagcccagca tccgggccca gacacttgat ggatccacac 1561 atattcactt ccaactttaa caatggcatt ggaaggcata agacctacct gtgctacgaa 1621 gtggagcgcc tggacaatgg cacctcggtc aagatggacc agcacagggg ctttctacac 1681 aaccaggcta agaatcttct ctgtggcttt tacggccgcc atgcggagct gcgcttcttg 1741 gacctggttc cttctttgca gttggacccg gcccagatct acagggtcac ttggttcatc 1801 tcctggagcc cctgcttctc ctggggctgt gccggggaag tgcgtgcgtt ccttcaggag 1861 aacacacacg tgagactgcg tatcttcgct gcccgcatct atgattacga ccccctatat 1921 aaggaggcac tgcaaatgct gcgggatgct ggggcccaag tctccatcat gacctacgat 1981 gaatttaagc actgctggga cacctttgtg gaccaccagg gatgtccctt ccagccctgg 2041 gatggactag atgagcacag ccaagccctg agtgggaggc tgcgggccat tctccagaat 2101 cagggaaact ctggttctga aactccaggt acttctgaat ctgctactcc agaatctatg 2161 gacaagaagt actccattgg gctcgctatc ggcacaaaca gcgtcggttg ggccgtcatt 2221 acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc tgggcaatac cgatcgccac 2281 agcataaaga agaacctcat tggcgccctc ctgttcgact ccggggagac ggccgaagcc 2341 acgcggctca aaagaacagc acggcgcaga tatacccgca gaaagaatcg gatctgctac 2401 ctgcaggaga tctttagtaa tgagatggct aaggtggatg actctttctt ccataggctg 2461 gaggagtcct ttttggtgga ggaggataaa aagcacgagc gccacccaat ctttggcaat 2521 atcgtggacg aggtggcgta ccatgaaaag tacccaacca tatatcatct gaggaagaag 2581 cttgtagaca gtactgataa ggctgacttg cggttgatct atctcgcgct ggcgcatatg 2641 atcaaatttc ggggacactt cctcattgag ggggacctga acccagacaa cagcgatgtc 2701 gacaaactct ttatccaact ggttcagact tacaatcagc ttttcgaaga gaacccgatc 2761 aacgcatccg gagttgacgc caaagcaatc ctgagcgcta ggctgtccaa atcccggcgg 2821 ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga acggcctgtt tggtaatctt 2881 atcgccctgt cactcgggct gacccccaac tttaaatcta acttcgacct ggccgaagat 2941 gccaagcttc aactgagcaa agacacctac gatgatgatc tcgacaatct gctggcccag 3001 atcggcgacc agtacgcaga cctttttttg gcggcaaaga acctgtcaga cgccattctg 3061 ctgagtgata ttctgcgagt gaacacggag atcaccaaag ctccgctgag cgctagtatg 3121 atcaagcgct atgatgagca ccaccaagac ttgactttgc tgaaggccct tgtcagacag 3181 caactgcctg agaagtacaa ggaaattttc ttcgatcagt ctaaaaatgg ctacgccgga 3241 tacattgacg gcggagcaag ccaggaggaa ttttacaaat ttattaagcc catcttggaa 3301 aaaatggacg gcaccgagga gctgctggta aagcttaaca gagaagatct gttgcgcaaa 3361 cagcgcactt tcgacaatgg aagcatcccc caccagattc acctgggcga actgcacgct 3421 atcctcaggc ggcaagagga tttctacccc tttttgaaag ataacaggga aaagattgag 3481 aaaatcctta catttcggat accctactat gtaggccccc tcgcccgggg aaattccaga 3541 ttcgcgtgga tgactcgcaa atcagaagag accatcactc cctggaactt cgaggaagtc 3601 gtggataagg gggcctctgc ccagtccttc atcgaaagga tgactaactt tgataaaaat 3661 ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt acgagtactt cacagtttat 3721 aacgagctca ccaaggtcaa atacgtcaca gaagggatga gaaagccagc attcctgtct 3781 ggagagcaga agaaagctat cgtggacctc ctcttcaaga cgaaccggaa agttaccgtg 3841 aaacagctca aagaagacta tttcaaaaag attgaatgtt tcgactctgt tgaaatcagc 3901 ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc acgatctcct gaaaatcatt 3961 aaagacaagg acttcctgga caatgaggag aacgaggaca ttcttgagga cattgtcctc 4021 acccttacgt tgtttgaaga tagggagatg attgaagaac gcttgaaaac ttacgctcat 4081 ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc gatatacagg atgggggcgg 4141 ctgtcaagaa aactgatcaa tgggatccga gacaagcaga gtggaaagac aatcctggat 4201 tttcttaagt ccgatggatt tgccaaccgg aacttcatgc agttgatcca tgatgactct 4261 ctcaccttta aggaggacat ccagaaagca caagtttctg gccaggggga cagtcttcac 4321 gagcacatcg ctaatcttgc aggtagccca gctatcaaaa agggaatact gcagaccgtt 4381 aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata agcccgagaa tatcgttatc 4441 gagatggccc gagagaacca aactacccag aagggacaga agaacagtag ggaaaggatg 4501 aagaggattg aagagggtat aaaagaactg gggtcccaaa tccttaagga acacccagtt 4561 gaaaacaccc agcttcagaa tgagaagctc tacctgtact acctgcagaa cggcagggac 4621 atgtacgtgg atcaggaact ggacatcaat cggctctccg actacgacgt ggatcatatc 4681 gtgccccagt cttttctcaa agatgattct attgataata aagtgttgac aagatccgat 4741 aaaaatagag ggaagagtga taacgtcccc tcagaagaag ttgtcaagaa aatgaaaaat 4801 tattggcggc agctgctgaa cgccaaactg atcacacaac ggaagttcga taatctgact 4861 aaggctgaac gaggtggcct gtctgagttg gataaagccg gcttcatcaa aaggcagctt 4921 gttgagacac gccagatcac caagcacgtg gcccaaattc tcgattcacg catgaacacc 4981 aagtacgatg aaaatgacaa actgattcga gaggtgaaag ttattactct gaagtctaag 5041 ctggtctcag atttcagaaa ggactttcag ttttataagg tgagagagat caacaattac 5101 caccatgcgc atgatgccta cctgaatgca gtggtaggca ctgcacttat caaaaaatat 5161 cccaagcttg aatctgaatt tgtttacgga gactataaag tgtacgatgt taggaaaatg 5221 atcgcaaagt ctgagcagga aataggcaag gccaccgcta agtacttctt ttacagcaat 5281 attatgaatt ttttcaagac cgagattaca ctggccaatg gagagattcg gaagcgacca 5341 cttatcgaaa caaacggaga aacaggagaa atcgtgtggg acaagggtag ggatttcgcg 5401 acagtccgga aggtcctgtc catgccgcag gtgaacatcg ttaaaaagac cgaagtacag 5461 accggaggct tctccaagga aagtatcctc ccgaaaagga acagcgacaa gctgatcgca 5521 cgcaaaaaag attgggaccc caagaaatac ggcggattcg attctcctac agtcgcttac 5581 agtgtactgg ttgtggccaa agtggagaaa gggaagtcta aaaaactcaa aagcgtcaag 5641 gaactgctgg gcatcacaat catggagcga tcaagcttcg aaaaaaaccc catcgacttt 5701 ctcgaggcga aaggatataa agaggtcaaa aaagacctca tcattaagct tcccaagtac 5761 tctctctttg agcttgaaaa cggccggaaa cgaatgctcg ctagtgcggg cgagctgcag 5821 aaaggtaacg agctggcact gccctctaaa tacgttaatt tcttgtatct ggccagccac 5881 tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga agcagctgtt cgtggaacaa 5941 cacaaacact accttgatga gatcatcgag caaataagcg aattctccaa aagagtgatc 6001 ctcgccgacg ctaacctcga taaggtgctt tctgcttaca ataagcacag ggataagccc 6061 atcagggagc aggcagaaaa cattatccac ttgtttactc tgaccaactt gggcgcgcct 6121 gcagccttca agtacttcga caccaccata gacagaaagc ggtacacctc tacaaaggag 6181 gtcctggacg ccacactgat tcatcagtca attacggggc tctatgaaac aagaatcgac 6241 ctctctcagc tcggtggaga ctctggtggt tcaactaatt tatctgatat tattgaaaaa 6301 gaaacaggta aacaattagt tattcaagaa tcaattttaa tgttgccaga agaagttgaa 6361 gaagttattg gtaataaacc agaatctgat attttagttc atactgctta tgatgaatct 6421 actgatgaaa atgttatgtt attgacttca gatgctccag aatataaacc atgggcttta 6481 gttattcaag attctaatgg tgaaaataaa attaaaatgt tgtctggtgg ttcacccaag 6541 aagaagagga aggtgtgatc tcttctcgag tcatgtaatt agttatgtca cgcttacatt 6601 cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct 6661 aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa 6721 tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc 6781 ttgagaaggt tttgggacgc tcgaaggctt taatttgcgg ccggtaccca attcgcccta 6841 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa 6901 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa 6961 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg 7021 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 7081 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 7141 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt 7201 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg 7261 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt 7321 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta 7381 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt 7441 aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta 7501 cgcatctgtg cggtatttta caccgcatat cgaccggtcg aggagaactt ctagtatatc 7561 tacataccta atattattgc cttattaaaa atggaatccc aacaattaca tcaaaatcca 7621 cattctcttc aaaatcaatt gtcctgtact tccttgttca tgtgtgttca aaaacgttat 7681 atttatagga taattatact ctatttctca acaagtaatt ggttgtttgg ccgagcggtc 7741 taaggcgcct gattcaagaa atatcttgac cgcagttaac tgtgggaata ctcaggtatc 7801 gtaagatgca agagttcgaa tctcttagca accattattt ttttcctcaa cataacgaga 7861 acacacaggg gcgctatcgc acagaatcaa attcgatgac tggaaatttt ttgttaattt 7921 cagaggtcgc ctgacgcata tacctttttc aactgaaaaa ttgggagaaa aaggaaaggt 7981 gagagcgccg gaaccggctt ttcatataga atagagaagc gttcatgact aaatgcttgc 8041 atcacaatac ttgaagttga caatattatt taaggaccta ttgttttttc caataggtgg 8101 ttagcaatcg tcttactttc taacttttct taccttttac atttcagcaa tatatatata 8161 tatatttcaa ggatatacca ttctaatgtc tgcccctaag aagatcgtcg ttttgccagg 8221 tgaccacgtt ggtcaagaaa tcacagccga agccattaag gttcttaaag ctatttctga 8281 tgttcgttcc aatgtcaagt tcgatttcga aaatcattta attggtggtg ctgctatcga 8341 tgctacaggt gttccacttc cagatgaggc gctggaagcc tccaagaagg ctgatgccgt 8401 tttgttaggt gctgtgggtg gtcctaaatg gggtaccggt agtgttagac ctgaacaagg 8461 tttactaaaa atccgtaaag aacttcaatt gtacgccaac ttaagaccat gtaactttgc 8521 atccgactct cttttagact tatctccaat caagccacaa tttgctaaag gtactgactt 8581 cgttgttgtc agagaattag tgggaggtat ttactttggt aagagaaagg aagacgatgg 8641 tgatggtgtc gcttgggata gtgaacaata caccgttcca gaagtgcaaa gaatcacaag 8701 aatggccgct ttcatggccc tacaacatga gccaccattg cctatttggt ccttggataa 8761 agctaatgtt ttggcctctt caagattatg gagaaaaact gtggaggaaa ccatcaagaa 8821 cgaattccct acattgaagg ttcaacatca attgattgat tctgccgcca tgatcctagt 8881 taagaaccca acccacctaa atggtattat aatcaccagc aacatgtttg gtgatatcat 8941 ctccgatgaa gcctccgtta tcccaggttc cttgggtttg ttgccatctg cgtccttggc 9001 ctctttgcca gacaagaaca ccgcatttgg tttgtacgaa ccatgccacg gttctgctcc 9061 agatttgcca aagaataagg tcaaccctat cgccactatc ttgtctgctg caatgatgtt 9121 gaaattgtca ttgaacttgc ctgaagaagg taaggccatt gaagatgcag ttaaaaaggt 9181 tttggatgca ggtatcagaa ctggtgattt aggtggttcc aacagtacca ccgaagtcgg 9241 tgatgctgtc gccgaagaag ttaagaaaat ccttgcttaa aaagattctc tttttttatg 9301 atatttgtac ataaacttta taaatgaaat tcataataga aacgacacga aattacaaaa 9361 tggaatatgt tcatagggta gacgaaacta tatacgcaat ctacatacat ttatcaagaa 9421 ggagaaaaag gaggatgtaa aggaatacag gtaagcaaat tgatactaat ggctcaacgt 9481 gataaggaaa aagaattgca ctttaacatt aatattgaca aggaggaggg caccacacaa 9541 aaagttaggt gtaacagaaa atcatgaaac tatgattcct aatttatata ttggaggatt 9601 ttctctaaaa aaaaaaaaat acaacaaata aaaaacactc aatgacctga ccatttgatg 9661 gagtttaagt caataccttc ttgaaccatt tcccataatg gtgaaagttc cctcaagaat 9721 tttactctgt cagaaacggc cttaacgacg tagtcgatat ggtgcactct cagtacaatc 9781 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc 9841 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc 9901 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg 9961 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagga cggatcgctt 10021 gcctgtaact tacacgcgcc tcgtatcttt taatgatgga ataatttggg aatttactct 10081 gtgtttattt atttttatgt tttgtatttg gattttagaa agtaaataaa gaaggtagaa 10141 gagttacgga atgaagaaaa aaaaataaac aaaggtttaa aaaatttcaa caaaaagcgt 10201 actttacata tatatttatt agacaagaaa agcagattaa atagatatac attcgattaa 10261 cgataagtaa aatgtaaaat cacaggattt tcgtgtgtgg tcttctacac agacaagatg 10321 aaacaattcg gcattaatac ctgagagcag gaagagcaag ataaaaggta gtatttgttg 10381 gcgatccccc tagagtcttt tacatcttcg gaaaacaaaa actatttttt ctttaatttc 10441 tttttttact ttctattttt aatttatata tttatattaa aaaatttaaa ttataattat 10501 ttttatagca cgtgatgaaa aggacccagg tggcactttt cggggaaatg tgcgcggaac 10561 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc 10621 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt 10681 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct 10741 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga 10801 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag 10861 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca 10921 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga 10981 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag 11041 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc 11101 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa 11161 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt 11221 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg 11281 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt 11341 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg 11401 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat 11461 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact 11521 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa 11581 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt 11641 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttc //