LOCUS Exported 11778 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses BE3 in yeast cells. ACCESSION . VERSION . KEYWORDS pJT25_GalL_BE3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11778) 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 11778) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11778 /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..2199 /codon_start=1 /product="cytidine deaminase (C to U editing enzyme) from rat" /label=APOBEC-1 /note="can use ssDNA as a substrate (Komor et al., 2016)" /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI LWATGLK" CDS 2248..6351 /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 6364..6612 /codon_start=1 /product="uracil-DNA glycosylase inhibitor from a Bacillus subtilis bacteriophage (Mol et al., 1995)" /label=UGI /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML" CDS 6625..6645 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" terminator 6661..6908 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(6677..6695) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" primer_bind complement(6926..6945) /label=T7 /note="T7 promoter, forward primer" promoter complement(6927..6945) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(6955..6972) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(6955..6971) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6964..6986) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 7112..7567 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7199..7218) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7409..7430 /label=F1ori-F /note="F1 origin, forward primer" promoter 7868..8275 /gene="S. cerevisiae LEU2" /label=LEU2 promoter CDS 8276..9370 /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(9864..9883) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 9983..10005 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(10043..10061) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" misc_feature 10103..10606 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" promoter 10643..10747 /gene="bla" /label=AmpR promoter CDS 10748..11608 /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(10966..10985) /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 ctctcctccg 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 aaatgagttc cgagacaggc cctgtagctg ttgatcccac tctgaggaga 1561 agaattgagc cccacgagtt tgaagtcttc tttgaccccc gggaacttcg gaaagagacc 1621 tgtctgctgt atgagatcaa ctggggagga aggcacagca tctggcgaca cacgagccaa 1681 aacaccaaca aacacgttga agtcaatttc atagaaaaat ttactacaga aagatacttt 1741 tgtccaaaca ccagatgctc cattacctgg ttcctgtcct ggagtccctg tggggagtgc 1801 tccagggcca ttacagaatt tttgagccga tacccccatg taactctgtt tatttatata 1861 gcacggcttt atcaccacgc agatcctcga aatcggcaag gactcaggga ccttattagc 1921 agcggtgtta ctatccagat catgacggag caagagtctg gctactgctg gaggaatttt 1981 gtcaactact ccccttcgaa tgaagctcat tggccaaggt acccccatct gtgggtgagg 2041 ctgtacgtac tggaactcta ctgcatcatt ttaggacttc caccctgttt aaatatttta 2101 agaagaaaac aacctcaact cacgtttttc acgattgctc ttcaaagctg ccattaccaa 2161 aggctaccac cccacatcct gtgggccaca gggttgaaat ctggttctga aactccaggt 2221 acttctgaat ctgctactcc agaatctatg gacaagaagt actccattgg gctcgctatc 2281 ggcacaaaca gcgtcggttg ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa 2341 ttcaaagttc tgggcaatac cgatcgccac agcataaaga agaacctcat tggcgccctc 2401 ctgttcgact ccggggagac ggccgaagcc acgcggctca aaagaacagc acggcgcaga 2461 tatacccgca gaaagaatcg gatctgctac ctgcaggaga tctttagtaa tgagatggct 2521 aaggtggatg actctttctt ccataggctg gaggagtcct ttttggtgga ggaggataaa 2581 aagcacgagc gccacccaat ctttggcaat atcgtggacg aggtggcgta ccatgaaaag 2641 tacccaacca tatatcatct gaggaagaag cttgtagaca gtactgataa ggctgacttg 2701 cggttgatct atctcgcgct ggcgcatatg atcaaatttc ggggacactt cctcatcgag 2761 ggggacctga acccagacaa cagcgatgtc gacaaactct ttatccaact ggttcagact 2821 tacaatcagc ttttcgaaga gaacccgatc aacgcatccg gagttgacgc caaagcaatc 2881 ctgagcgcta ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg 2941 gagaagaaga acggcctgtt tggtaatctt atcgccctgt cactcgggct gacccccaac 3001 tttaaatcta acttcgacct ggccgaagat gccaagcttc aactgagcaa agacacctac 3061 gatgatgatc tcgacaatct gctggcccag atcggcgacc agtacgcaga cctttttttg 3121 gcggcaaaga acctgtcaga cgccattctg ctgagtgata ttctgcgagt gaacacggag 3181 atcaccaaag ctccgctgag cgctagtatg atcaagcgct atgatgagca ccaccaagac 3241 ttgactttgc tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc 3301 ttcgatcagt ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa 3361 ttttacaaat ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta 3421 aagcttaaca gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc 3481 caccagattc acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc 3541 tttttgaaag ataacaggga aaagattgag aaaatcctca catttcggat accctactat 3601 gtaggccccc tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag 3661 accatcactc cctggaactt cgaggaagtc gtggataagg gggcctctgc ccagtccttc 3721 atcgaaagga tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac 3781 tctctgctgt acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca 3841 gaagggatga gaaagccagc attcctgtct ggagagcaga agaaagctat cgtggacctc 3901 ctcttcaaga cgaaccggaa agttaccgtg aaacagctca aagaagacta tttcaaaaag 3961 attgaatgtt tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg 4021 ggaacgtatc acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag 4081 aacgaggaca ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg 4141 attgaagaac gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaaacagctc 4201 aagaggcgcc gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tgggatccga 4261 gacaagcaga gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg 4321 aacttcatgc agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca 4381 caagtttctg gccaggggga cagtcttcac gagcacatcg ctaatcttgc aggtagccca 4441 gctatcaaaa agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg 4501 ggaaggcata agcccgagaa tatcgttatc gagatggccc gagagaacca aactacccag 4561 aagggacaga agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg 4621 gggtcccaaa tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc 4681 tacctgtact acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaat 4741 cggctctccg actacgacgt ggatcatatc gtgccccagt cttttctcaa agatgattct 4801 attgataata aagtgttgac aagatccgat aaaaatagag ggaagagtga taacgtcccc 4861 tcagaagaag ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg 4921 atcacacaac ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg 4981 gataaagccg gcttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg 5041 gcccaaattc tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga 5101 gaggtgaaag ttattactct gaagtctaag ctggtctcag atttcagaaa ggactttcag 5161 ttttataagg tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca 5221 gtggtaggca ctgcacttat caaaaaatat cccaagcttg aatctgaatt tgtttacgga 5281 gactataaag tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag 5341 gccaccgcta agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca 5401 ctggccaatg gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa 5461 atcgtgtggg acaagggtag ggatttcgcg acagtccgga aggtcctgtc catgccgcag 5521 gtgaacatcg ttaaaaagac cgaagtacag accggaggct tctccaagga aagtatcctc 5581 ccgaaaagga acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac 5641 ggcggattcg attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa 5701 gggaagtcta aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga 5761 tcaagcttcg aaaaaaaccc catcgacttt ctcgaggcga aaggatataa agaggtcaaa 5821 aaagacctca tcattaagct tcccaagtac tctctctttg agcttgaaaa cggccggaaa 5881 cgaatgctcg ctagtgcggg cgagctgcag aaaggtaacg agctggcact gccctctaaa 5941 tacgttaatt tcttgtatct ggccagccac tatgaaaagc tcaaagggtc tcccgaagat 6001 aatgagcaga agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag 6061 caaataagcg aattctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt 6121 tctgcttaca ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac 6181 ttgtttactc tgaccaactt gggcgcgcct gcagccttca agtacttcga caccaccata 6241 gacagaaagc ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca 6301 attacggggc tctatgaaac aagaatcgac ctctctcagc tcggtggaga ctctggtggt 6361 tcaactaatt tatctgatat tattgaaaaa gaaacaggta aacaattagt tattcaagaa 6421 tcaattttaa tgttgccaga agaagttgaa gaagttattg gtaataaacc agaatctgat 6481 attttagttc atactgctta tgatgaatct actgatgaaa atgttatgtt attgacttca 6541 gatgctccag aatataaacc atgggcttta gttattcaag attctaatgg tgaaaataaa 6601 attaaaatgt tgtctggtgg ttcacccaag aagaagagga aggtgtgatc tcttctcgag 6661 tcatgtaatt agttatgtca cgcttacatt cacgccctcc ccccacatcc gctctaaccg 6721 aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt 6781 tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt 6841 acgcatgtaa cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt 6901 taatttgcgg ccggtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc 6961 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc 7021 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc 7081 ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg 7141 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 7201 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 7261 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 7321 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 7381 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 7441 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg 7501 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt 7561 acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat 7621 cgaccggtcg aggagaactt ctagtatatc tacataccta atattattgc cttattaaaa 7681 atggaatccc aacaattaca tcaaaatcca cattctcttc aaaatcaatt gtcctgtact 7741 tccttgttca tgtgtgttca aaaacgttat atttatagga taattatact ctatttctca 7801 acaagtaatt ggttgtttgg ccgagcggtc taaggcgcct gattcaagaa atatcttgac 7861 cgcagttaac tgtgggaata ctcaggtatc gtaagatgca agagttcgaa tctcttagca 7921 accattattt ttttcctcaa cataacgaga acacacaggg gcgctatcgc acagaatcaa 7981 attcgatgac tggaaatttt ttgttaattt cagaggtcgc ctgacgcata tacctttttc 8041 aactgaaaaa ttgggagaaa aaggaaaggt gagagcgccg gaaccggctt ttcatataga 8101 atagagaagc gttcatgact aaatgcttgc atcacaatac ttgaagttga caatattatt 8161 taaggaccta ttgttttttc caataggtgg ttagcaatcg tcttactttc taacttttct 8221 taccttttac atttcagcaa tatatatata tatatttcaa ggatatacca ttctaatgtc 8281 tgcccctaag aagatcgtcg ttttgccagg tgaccacgtt ggtcaagaaa tcacagccga 8341 agccattaag gttcttaaag ctatttctga tgttcgttcc aatgtcaagt tcgatttcga 8401 aaatcattta attggtggtg ctgctatcga tgctacaggt gttccacttc cagatgaggc 8461 gctggaagcc tccaagaagg ctgatgccgt tttgttaggt gctgtgggtg gtcctaaatg 8521 gggtaccggt agtgttagac ctgaacaagg tttactaaaa atccgtaaag aacttcaatt 8581 gtacgccaac ttaagaccat gtaactttgc atccgactct cttttagact tatctccaat 8641 caagccacaa tttgctaaag gtactgactt cgttgttgtc agagaattag tgggaggtat 8701 ttactttggt aagagaaagg aagacgatgg tgatggtgtc gcttgggata gtgaacaata 8761 caccgttcca gaagtgcaaa gaatcacaag aatggccgct ttcatggccc tacaacatga 8821 gccaccattg cctatttggt ccttggataa agctaatgtt ttggcctctt caagattatg 8881 gagaaaaact gtggaggaaa ccatcaagaa cgaattccct acattgaagg ttcaacatca 8941 attgattgat tctgccgcca tgatcctagt taagaaccca acccacctaa atggtattat 9001 aatcaccagc aacatgtttg gtgatatcat ctccgatgaa gcctccgtta tcccaggttc 9061 cttgggtttg ttgccatctg cgtccttggc ctctttgcca gacaagaaca ccgcatttgg 9121 tttgtacgaa ccatgccacg gttctgctcc agatttgcca aagaataagg tcaaccctat 9181 cgccactatc ttgtctgctg caatgatgtt gaaattgtca ttgaacttgc ctgaagaagg 9241 taaggccatt gaagatgcag ttaaaaaggt tttggatgca ggtatcagaa ctggtgattt 9301 aggtggttcc aacagtacca ccgaagtcgg tgatgctgtc gccgaagaag ttaagaaaat 9361 ccttgcttaa aaagattctc tttttttatg atatttgtac ataaacttta taaatgaaat 9421 tcataataga aacgacacga aattacaaaa tggaatatgt tcatagggta gacgaaacta 9481 tatacgcaat ctacatacat ttatcaagaa ggagaaaaag gaggatgtaa aggaatacag 9541 gtaagcaaat tgatactaat ggctcaacgt gataaggaaa aagaattgca ctttaacatt 9601 aatattgaca aggaggaggg caccacacaa aaagttaggt gtaacagaaa atcatgaaac 9661 tatgattcct aatttatata ttggaggatt ttctctaaaa aaaaaaaaat acaacaaata 9721 aaaaacactc aatgacctga ccatttgatg gagtttaagt caataccttc ttgaaccatt 9781 tcccataatg gtgaaagttc cctcaagaat tttactctgt cagaaacggc cttaacgacg 9841 tagtcgatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc 9901 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct 9961 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca 10021 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg 10081 ataataatgg tttcttagga cggatcgctt gcctgtaact tacacgcgcc tcgtatcttt 10141 taatgatgga ataatttggg aatttactct gtgtttattt atttttatgt tttgtatttg 10201 gattttagaa agtaaataaa gaaggtagaa gagttacgga atgaagaaaa aaaaataaac 10261 aaaggtttaa aaaatttcaa caaaaagcgt actttacata tatatttatt agacaagaaa 10321 agcagattaa atagatatac attcgattaa cgataagtaa aatgtaaaat cacaggattt 10381 tcgtgtgtgg tcttctacac agacaagatg aaacaattcg gcattaatac ctgagagcag 10441 gaagagcaag ataaaaggta gtatttgttg gcgatccccc tagagtcttt tacatcttcg 10501 gaaaacaaaa actatttttt ctttaatttc tttttttact ttctattttt aatttatata 10561 tttatattaa aaaatttaaa ttataattat ttttatagca cgtgatgaaa aggacccagg 10621 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc 10681 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag 10741 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg 10801 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt 10861 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt 10921 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt 10981 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa 11041 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag 11101 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac 11161 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac 11221 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac 11281 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac 11341 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact 11401 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg 11461 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt 11521 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat 11581 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta 11641 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa 11701 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga 11761 aaagatcaaa ggatcttc //