LOCUS Exported 12061 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses cCDA1-BE3 in yeast cells. ACCESSION . VERSION . KEYWORDS pJT46_GalL_cCDA1-BE3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12061) AUTHORS Tan J, Zhang F, Karcher D, Bock R TITLE Engineering of high-precision base editors for site-specific single nucleotide replacement. JOURNAL Nat Commun. 2019 Jan 25;10(1):439. doi: 10.1038/s41467-018-08034-8. PUBMED 30683865 REFERENCE 2 (bases 1 to 12061) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12061 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..442 /gene="S. cerevisiae GAL1" /label=GAL1 promoter /note="inducible promoter, regulated by Gal4" protein_bind 1..118 /label=UAS /bound_moiety="Gal4" /note="upstream activating sequence mediating Gal4-dependent induction" primer_bind 413..436 /label=GAL1 /note="S. cerevisiae GAL1 promoter, forward primer" CDS 452..4555 /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 4604..5227 /codon_start=1 /product="cytidine deaminase from the sea lamprey Petromyzon marinus (Rogozin et al., 2007)" /label=PmCDA1 /translation="MTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGER RACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKI LEWYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIF IQSSHNQLNENRWLEKTLKRAEKRRSELSIMIQVKILHTTKSPAV" CDS 5246..5494 /codon_start=1 /product="uracil-DNA glycosylase inhibitor from a Bacillus subtilis bacteriophage (Mol et al., 1995)" /label=UGI /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML" CDS 5507..5527 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" terminator 5561..5748 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(5616..5640) /label=pHybLex-R /note="ADH terminator, reverse primer" terminator 5785..5830 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind complement(5866..5885) /label=T7 /note="T7 promoter, forward primer" promoter complement(5867..5885) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5892..5909) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(5892..5908) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5901..5923) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 6049..6504 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6136..6155) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6346..6367 /label=F1ori-F /note="F1 origin, forward primer" promoter 6805..7212 /gene="S. cerevisiae LEU2" /label=LEU2 promoter CDS 7213..8307 /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(8801..8820) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 8920..8942 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(8980..8998) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" misc_feature 9040..9543 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" promoter 9580..9684 /gene="bla" /label=AmpR promoter CDS 9685..10545 /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(9903..9922) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 10716..11304 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 11205..11224 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 11458..11475 /label=L4440 /note="L4440 vector, forward primer" protein_bind 11592..11613 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 11628..11658 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 11666..11682 /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 11671..11693 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 11690..11706 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 11690..11706 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 11727..11745 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" ORIGIN 1 cggattagaa gccgccgagc gggtgacagc cctccgaagg aagactctcc tccgtgcgtc 61 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa 121 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc 181 tggccccaca aaccttcaaa tgaacgaatc aaattaacaa ccataggatg ataatgcgat 241 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt 301 aacagatata taaatgcaaa aactgcataa ccactttaac taatactttc aacattttcg 361 gtttgtatta cttcttattc aaatgtaata aaagtatcaa caaaaaattg ttaatatacc 421 tctatacttt aacgtcaagg agaaaaaacc catggacaag aagtactcca ttgggctcgc 481 tatcggcaca aacagcgtcg gttgggccgt cattacggac gagtacaagg tgccgagcaa 541 aaaattcaaa gttctgggca ataccgatcg ccacagcata aagaagaacc tcattggcgc 601 cctcctgttc gactccgggg agacggccga agccacgcgg ctcaaaagaa cagcacggcg 661 cagatatacc cgcagaaaga atcggatctg ctacctgcag gagatcttta gtaatgagat 721 ggctaaggtg gatgactctt tcttccatag gctggaggag tcctttttgg tggaggagga 781 taaaaagcac gagcgccacc caatctttgg caatatcgtg gacgaggtgg cgtaccatga 841 aaagtaccca accatatatc atctgaggaa gaagcttgta gacagtactg ataaggctga 901 cttgcggttg atctatctcg cgctggcgca tatgatcaaa tttcggggac acttcctcat 961 cgagggggac ctgaacccag acaacagcga tgtcgacaaa ctctttatcc aactggttca 1021 gacttacaat cagcttttcg aagagaaccc gatcaacgca tccggagttg acgccaaagc 1081 aatcctgagc gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg cacagctccc 1141 tggggagaag aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc 1201 caactttaaa tctaacttcg acctggccga agatgccaag cttcaactga gcaaagacac 1261 ctacgatgat gatctcgaca atctgctggc ccagatcggc gaccagtacg cagacctttt 1321 tttggcggca aagaacctgt cagacgccat tctgctgagt gatattctgc gagtgaacac 1381 ggagatcacc aaagctccgc tgagcgctag tatgatcaag cgctatgatg agcaccacca 1441 agacttgact ttgctgaagg cccttgtcag acagcaactg cctgagaagt acaaggaaat 1501 tttcttcgat cagtctaaaa atggctacgc cggatacatt gacggcggag caagccagga 1561 ggaattttac aaatttatta agcccatctt ggaaaaaatg gacggcaccg aggagctgct 1621 ggtaaagctt aacagagaag atctgttgcg caaacagcgc actttcgaca atggaagcat 1681 cccccaccag attcacctgg gcgaactgca cgctatcctc aggcggcaag aggatttcta 1741 cccctttttg aaagataaca gggaaaagat tgagaaaatc ctcacatttc ggatacccta 1801 ctatgtaggc cccctcgccc ggggaaattc cagattcgcg tggatgactc gcaaatcaga 1861 agagaccatc actccctgga acttcgagga agtcgtggat aagggggcct ctgcccagtc 1921 cttcatcgaa aggatgacta actttgataa aaatctgcct aacgaaaagg tgcttcctaa 1981 acactctctg ctgtacgagt acttcacagt ttataacgag ctcaccaagg tcaaatacgt 2041 cacagaaggg atgagaaagc cagcattcct gtctggagag cagaagaaag ctatcgtgga 2101 cctcctcttc aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag actatttcaa 2161 aaagattgaa tgtttcgact ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc 2221 cctgggaacg tatcacgatc tcctgaaaat cattaaagac aaggacttcc tggacaatga 2281 ggagaacgag gacattcttg aggacattgt cctcaccctt acgttgtttg aagataggga 2341 gatgattgaa gaacgcttga aaacttacgc tcatctcttc gacgacaaag tcatgaaaca 2401 gctcaagagg cgccgatata caggatgggg gcggctgtca agaaaactga tcaatgggat 2461 ccgagacaag cagagtggaa agacaatcct ggattttctt aagtccgatg gatttgccaa 2521 ccggaacttc atgcagttga tccatgatga ctctctcacc tttaaggagg acatccagaa 2581 agcacaagtt tctggccagg gggacagtct tcacgagcac atcgctaatc ttgcaggtag 2641 cccagctatc aaaaagggaa tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt 2701 aatgggaagg cataagcccg agaatatcgt tatcgagatg gcccgagaga accaaactac 2761 ccagaaggga cagaagaaca gtagggaaag gatgaagagg attgaagagg gtataaaaga 2821 actggggtcc caaatcctta aggaacaccc agttgaaaac acccagcttc agaatgagaa 2881 gctctacctg tactacctgc agaacggcag ggacatgtac gtggatcagg aactggacat 2941 caatcggctc tccgactacg acgtggatca tatcgtgccc cagtcttttc tcaaagatga 3001 ttctattgat aataaagtgt tgacaagatc cgataaaaat agagggaaga gtgataacgt 3061 cccctcagaa gaagttgtca agaaaatgaa aaattattgg cggcagctgc tgaacgccaa 3121 actgatcaca caacggaagt tcgataatct gactaaggct gaacgaggtg gcctgtctga 3181 gttggataaa gccggcttca tcaaaaggca gcttgttgag acacgccaga tcaccaagca 3241 cgtggcccaa attctcgatt cacgcatgaa caccaagtac gatgaaaatg acaaactgat 3301 tcgagaggtg aaagttatta ctctgaagtc taagctggtc tcagatttca gaaaggactt 3361 tcagttttat aaggtgagag agatcaacaa ttaccaccat gcgcatgatg cctacctgaa 3421 tgcagtggta ggcactgcac ttatcaaaaa atatcccaag cttgaatctg aatttgttta 3481 cggagactat aaagtgtacg atgttaggaa aatgatcgca aagtctgagc aggaaatagg 3541 caaggccacc gctaagtact tcttttacag caatattatg aattttttca agaccgagat 3601 tacactggcc aatggagaga ttcggaagcg accacttatc gaaacaaacg gagaaacagg 3661 agaaatcgtg tgggacaagg gtagggattt cgcgacagtc cggaaggtcc tgtccatgcc 3721 gcaggtgaac atcgttaaaa agaccgaagt acagaccgga ggcttctcca aggaaagtat 3781 cctcccgaaa aggaacagcg acaagctgat cgcacgcaaa aaagattggg accccaagaa 3841 atacggcgga ttcgattctc ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga 3901 gaaagggaag tctaaaaaac tcaaaagcgt caaggaactg ctgggcatca caattatgga 3961 gcgatcaagc ttcgaaaaaa accccatcga ctttctcgag gcgaaaggat ataaagaggt 4021 caaaaaagac ctcatcatta agcttcccaa gtactctctc tttgagcttg aaaacggccg 4081 gaaacgaatg ctcgctagtg cgggcgagct gcagaaaggt aacgagctgg cactgccctc 4141 taaatacgtt aatttcttgt atctggccag ccactatgaa aagctcaaag ggtctcccga 4201 agataatgag cagaagcagc tgttcgtgga acaacacaaa cactaccttg atgagatcat 4261 cgagcaaata agcgaattct ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt 4321 gctttctgct tacaataagc acagggataa gcccatcagg gagcaggcag aaaacattat 4381 ccacttgttt actctgacca acttgggcgc gcctgcagcc ttcaagtact tcgacaccac 4441 catagacaga aagcggtaca cctctacaaa ggaggtcctg gacgccacac tgattcatca 4501 gtcaattacg gggctctatg aaacaagaat cgacctctct cagctcggtg gagactctgg 4561 ttctgaaact ccaggtactt ctgaatctgc tactccagaa tctatgaccg acgctgagta 4621 cgtgagaatc catgagaagt tggacatcta cacgtttaag aaacagtttt tcaacaacaa 4681 aaaatccgtg tcgcatagat gctacgttct ctttgaatta aaacgacggg gtgaacgtag 4741 agcgtgtttt tggggctatg ctgtgaataa accacagagc gggacagaac gtggcattca 4801 cgccgaaatc tttagcatta gaaaagtcga agaatacctg cgcgacaacc ccggacaatt 4861 cacgataaat tggtactcat cctggagtcc ttgtgcagat tgcgctgaaa agatcttaga 4921 atggtataac caggagctgc gggggaacgg ccacactttg aaaatctggg cttgcaaact 4981 ctattacgag aaaaatgcga ggaatcaaat tgggctgtgg aacctcagag ataacggggt 5041 tgggttgaat gtaatggtaa gtgaacacta ccaatgttgc aggaaaatat tcatccaatc 5101 gtcgcacaat caattgaatg agaatagatg gcttgagaag actttgaagc gagctgaaaa 5161 acgacggagc gagttgtcca ttatgattca ggtaaaaata ctccacacca ctaagagtcc 5221 tgctgtttct agatctggtg gttcaactaa tttatctgat attattgaaa aagaaacagg 5281 taaacaatta gttattcaag aatcaatttt aatgttgcca gaagaagttg aagaagttat 5341 tggtaataaa ccagaatctg atattttagt tcatactgct tatgatgaat ctactgatga 5401 aaatgttatg ttattgactt cagatgctcc agaatataaa ccatgggctt tagttattca 5461 agattctaat ggtgaaaata aaattaaaat gttgtctggt ggttcaccca agaagaagag 5521 gaaggtgtga tctagaggct ccggataagg atcctaataa gcgaatttct tatgatttat 5581 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc 5641 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct 5701 ttctcaggta tagcatgagg tcgctcttat tgaccacacc tctaccggca tgccgagcaa 5761 atgcctgcaa atcgctcccg ggcaaaaaac cccctcaaga cccgtttaga ggccccaagg 5821 ggttatgcta tgcataccaa gcttccgcgg tggagctcca attcgcccta tagtgagtcg 5881 tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc 5941 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc 6001 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac gcgccctgta 6061 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca 6121 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct 6181 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc 6241 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat 6301 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc 6361 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc 6421 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta 6481 acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg 6541 tatttcacac cgcatatcga ccggtcgagg agaacttcta gtatatctac atacctaata 6601 ttattgcctt attaaaaatg gaatcccaac aattacatca aaatccacat tctcttcaaa 6661 atcaattgtc ctgtacttcc ttgttcatgt gtgttcaaaa acgttatatt tataggataa 6721 ttatactcta tttctcaaca agtaattggt tgtttggccg agcggtctaa ggcgcctgat 6781 tcaagaaata tcttgaccgc agttaactgt gggaatactc aggtatcgta agatgcaaga 6841 gttcgaatct cttagcaacc attatttttt tcctcaacat aacgagaaca cacaggggcg 6901 ctatcgcaca gaatcaaatt cgatgactgg aaattttttg ttaatttcag aggtcgcctg 6961 acgcatatac ctttttcaac tgaaaaattg ggagaaaaag gaaaggtgag agcgccggaa 7021 ccggcttttc atatagaata gagaagcgtt catgactaaa tgcttgcatc acaatacttg 7081 aagttgacaa tattatttaa ggacctattg ttttttccaa taggtggtta gcaatcgtct 7141 tactttctaa cttttcttac cttttacatt tcagcaatat atatatatat atttcaagga 7201 tataccattc taatgtctgc ccctaagaag atcgtcgttt tgccaggtga ccacgttggt 7261 caagaaatca cagccgaagc cattaaggtt cttaaagcta tttctgatgt tcgttccaat 7321 gtcaagttcg atttcgaaaa tcatttaatt ggtggtgctg ctatcgatgc tacaggtgtt 7381 ccacttccag atgaggcgct ggaagcctcc aagaaggctg atgccgtttt gttaggtgct 7441 gtgggtggtc ctaaatgggg taccggtagt gttagacctg aacaaggttt actaaaaatc 7501 cgtaaagaac ttcaattgta cgccaactta agaccatgta actttgcatc cgactctctt 7561 ttagacttat ctccaatcaa gccacaattt gctaaaggta ctgacttcgt tgttgtcaga 7621 gaattagtgg gaggtattta ctttggtaag agaaaggaag acgatggtga tggtgtcgct 7681 tgggatagtg aacaatacac cgttccagaa gtgcaaagaa tcacaagaat ggccgctttc 7741 atggccctac aacatgagcc accattgcct atttggtcct tggataaagc taatgttttg 7801 gcctcttcaa gattatggag aaaaactgtg gaggaaacca tcaagaacga gttccctaca 7861 ttgaaggttc aacatcaatt gattgattct gccgccatga tcctagttaa gaacccaacc 7921 cacctaaatg gtattataat caccagcaac atgtttggtg atatcatctc cgatgaagcc 7981 tccgttatcc caggttcctt gggtttgttg ccatctgcgt ccttggcctc tttgccagac 8041 aagaacaccg catttggttt gtacgaacca tgccacggtt ctgctccaga tttgccaaag 8101 aataaggtca accctatcgc cactatcttg tctgctgcaa tgatgttgaa attgtcattg 8161 aacttgcctg aagaaggtaa ggccattgaa gatgcagtta aaaaggtttt ggatgcaggt 8221 atcagaactg gtgatttagg tggttccaac agtaccaccg aagtcggtga tgctgtcgcc 8281 gaagaagtta agaaaatcct tgcttaaaaa gattctcttt ttttatgata tttgtacata 8341 aactttataa atgaaattca taatagaaac gacacgaaat tacaaaatgg aatatgttca 8401 tagggtagac gaaactatat acgcaatcta catacattta tcaagaagga gaaaaaggag 8461 gatgtaaagg aatacaggta agcaaattga tactaatggc tcaacgtgat aaggaaaaag 8521 aattgcactt taacattaat attgacaagg aggagggcac cacacaaaaa gttaggtgta 8581 acagaaaatc atgaaactat gattcctaat ttatatattg gaggattttc tctaaaaaaa 8641 aaaaaataca acaaataaaa aacactcaat gacctgacca tttgatggag tttaagtcaa 8701 taccttcttg aaccatttcc cataatggtg aaagttccct caagaatttt actctgtcag 8761 aaacggcctt aacgacgtag tcgatatggt gcactctcag tacaatctgc tctgatgccg 8821 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc 8881 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga 8941 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt 9001 tataggttaa tgtcatgata ataatggttt cttaggacgg atcgcttgcc tgtaacttac 9061 acgcgcctcg tatcttttaa tgatggaata atttgggaat ttactctgtg tttatttatt 9121 tttatgtttt gtatttggat tttagaaagt aaataaagaa ggtagaagag ttacggaatg 9181 aagaaaaaaa aataaacaaa ggtttaaaaa atttcaacaa aaagcgtact ttacatatat 9241 atttattaga caagaaaagc agattaaata gatatacatt cgattaacga taagtaaaat 9301 gtaaaatcac aggattttcg tgtgtggtct tctacacaga caagatgaaa caattcggca 9361 ttaatacctg agagcaggaa gagcaagata aaaggtagta tttgttggcg atccccctag 9421 agtcttttac atcttcggaa aacaaaaact attttttctt taatttcttt ttttactttc 9481 tatttttaat ttatatattt atattaaaaa atttaaatta taattatttt tatagcacgt 9541 gatgaaaagg acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta 9601 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt 9661 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc 9721 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa 9781 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt 9841 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt 9901 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc 9961 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg 10021 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg 10081 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac 10141 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca 10201 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta 10261 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat 10321 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa 10381 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag 10441 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat 10501 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt 10561 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg 10621 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga 10681 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta 10741 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa 10801 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact 10861 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca 10921 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt 10981 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg 11041 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag 11101 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta 11161 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat 11221 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg 11281 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc 11341 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac 11401 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc 11461 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aactgcctct ccccgcgcgt 11521 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag 11581 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg 11641 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc 11701 tatgaccatg attacgccaa gctcgaaatt aaccctcact aaagggaaca aaagctggta 11761 ccgggcccga attctctcct tctcttaggt ggcagagcag gtggagggtc gaccatacta 11821 gtttcaaaaa ttcttacttt ttttttggat ggacgcaaag aagtttaata atcatattac 11881 atggcattac caccatatac atatccatat acatatccat atacatatcc atatacatat 11941 ccatatctaa tcttacttat atgttgtgga aatgtaaaga gccccattat cttagcctaa 12001 aaaaaccttc tctttggaac tttcagtaat acgcttaact gctcattgct atattgaagt 12061 a //