LOCUS Exported 13805 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses ABEd CRISPR base editor and gRNA scaffold in Streptomyces. ACCESSION . VERSION . KEYWORDS pSABEd SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13805) AUTHORS Zhong Z, Guo J, Deng L, Chen L, Wang J, Li S, Xu W, Deng Z, Sun Y TITLE Base editing in Streptomyces with Cas9-deaminase fusions JOURNAL BioRxiv 630137 REFERENCE 2 (bases 1 to 13805) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13805 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(250..270) /label=T3 /note="T3 promoter, forward primer" promoter complement(250..268) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(297..315) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 383..487 /gene="bla" /label=AmpR promoter CDS 488..1348 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(706..725) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1519..2107 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2008..2027 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2261..2278 /label=L4440 /note="L4440 vector, forward primer" polyA_signal 2492..2626 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2516..2535) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2570..2589 /label=SV40pA-R /note="SV40 polyA, reverse primer" promoter 5844..6145 /label=rpsL promoter /note="promoter for the Xylanimonas cellulosilytica ribosomal protein S12 gene" CDS 7337..11440 /codon_start=1 /product="catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=dCas9 /note="RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain" /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" oriT 11803..11912 /note="incP origin of transfer" CDS 11945..12316 /codon_start=1 /gene="traJ" /product="oriT-recognizing protein" /label=traJ /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE KQDELGKVMMGVVRPRAEP" misc_RNA 12472..12547 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" CDS complement(13061..56) /codon_start=1 /gene="aac(3)IV" /product="aminoglycoside 3-N-acetyltransferase type IV " /label=ApmR /note="confers resistance to apramycin (Paget and Davies, 1996)" /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG" ORIGIN 1 agctcggctt ttcgccattc gtattgcacg acattgcact ccaccgctga tgacatcagt 61 cgatcatagc acgatcaacg gcactgttgc aaatagtcgg tggtgataaa cttatcatcc 121 ccttttgctg atggagctgc acatgaaccc attcaaaggc cggcattttc agcgtgacat 181 cattctgtgg gccgtacgct ggtactgcaa atacggcatc agttaccgtg agctgcaggt 241 cgacggatcc ctttagtgag ggttaattgc ggccgcgaat tcttgaagac gaaagggcct 301 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg 361 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc 421 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag 481 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg 541 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt 601 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt 661 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt 721 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa 781 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag 841 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac 901 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac 961 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac 1021 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac 1081 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact 1141 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg 1201 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt 1261 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat 1321 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta 1381 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa 1441 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga 1501 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac 1561 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt 1621 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc 1681 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat 1741 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag 1801 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc 1861 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag 1921 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac 1981 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg 2041 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct 2101 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc 2161 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga 2221 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga 2281 agcggaagag cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca 2341 ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc 2401 gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca 2461 acgacaggag cacgatcatg cgcacccgtc agatccagac atgataagat acattgatga 2521 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga 2581 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttcccg cacgatcagc 2641 gtcccgccga ccaggagcag tgcggccagc actgccgcta acgcctggtc ctggtcccgg 2701 tcctggtggt gcatcagtcc tccccgtgat cacttcggca cccaccgtag tgatcacccc 2761 cgacagcgga tcaaggggtt tgcgggtccc ggtcggcgcc gggcggggga ggcaggagcc 2821 gccgacgctg cctctgggac gggccggacg gcagggggac cggcggccgg gcgagctgca 2881 gccgggggtc cggcagggcc ggagcgggcg gaaccgtgct ctgacctgcg gcccgagttt 2941 cgtcacgtga cggaatggaa ggctgctgca tttcgtcacg tgacgtatct cggcgagcga 3001 ctgccgacgc cacggcggac acgatcgcct cgcgctggcg ccgggcctcg tacgcccgct 3061 ggcggcagga gcggcggcag tagtcccggc tccggccgac gccggattgc ttgatctccg 3121 agccgcacca ggcgcagagc ttcgcgccgt cggcgtccct gggggtggtg gtgctcatgg 3181 ccgacgaccg tacgcggcac gtctcgtagc gaggcgagtc gggcgcgagg taccgcctgc 3241 acgaagtgcc ggcggggccg accccgggcg agtaatccca ggattactcc cgcggcttcg 3301 accccggccg ccgtcgccgc gtacgtcacc gacccccgcc gtacgtcacc gggatgacgt 3361 acggcggggg ggagcgagtt agtgcgaagt gggcccactt gcgagccggg cgatgtgccg 3421 ggcggcccgc tcctggcggt cgtcggcgtc gtcgtcctgg tcgtcgtcct gctctcgccg 3481 tcggcgtgca gttgcttcct cgcggcgctg ggcgagggcg gcgagcatgt cggcgtacgc 3541 ctcggccacc tcccccgccg tgagcaccac cactgtgtcg gccgcgtcgg ccagcgccag 3601 gacctcccgc acccgttcgc ccacggccgc cgaatcctcg ttgccgtcct tgccttcggc 3661 ggcccgggtc gcctcgaggt cgagggcgcg gcgggtgacc gcgtgccatc cgtcctcggt 3721 cacggcgacc ccggcccgca gctccccgcc gtcggcgtcg gccgccagga gcagatcgag 3781 gtcgtcggcc tcggtgtcgc cgccgtcgag cccgagcatc tgccgcaggt agcgggtcca 3841 ttcgatggcc cggcgtcccc gggttgcccg ctcgtactcg tgccagcgcg agaggttcca 3901 ctccagcgag ccgaccccgg cggcgtcgtc ctcggtcatg ccgccggtca ggtccccgat 3961 ccgtccgagg agttcgaacg gggcgacgtt cccgccggtc gccgtcttga ggtcggcgcg 4021 ggcgagttcg agggcgggcg ccttcccgtc ctgggtcttg gcgatgtact cggcgaggtc 4081 gttggcgtcg cgctcggtct ccagccgctt gaagtcgacg ccgtgccggt cgtcgggcgt 4141 gaaggcgggg ttgaccttgc gcagggcggc ggtccacacg gaccgccagt gcccctgcca 4201 ctcgtcgagc gcggcgccgg tcggctcgaa ggtggcgacg atctgcttcg cggaccgctc 4261 cccctcggtc cggccgccga ccaggacgat cgcgtggatg tgcgggtgcc agccgttgat 4321 ctgccccacg gtgacttcgg tcgcgcggat catgccgacg tacccgatcc ggtctcggat 4381 gccctcgcgg tcggcggccc ggtgcccgtc cttggcccgg cgtccggccc acgtgccgcc 4441 cgtgatcagt cgctggtagg cgcccggccg ccgggggctg tccggcgtct tccgggtgcc 4501 ctggagggcg tccatgaggt ccgcgagccg gtccgtgtgc ccatggcggg ccgtgaaggt 4561 gaccaggtag gcggtccccc cgcgcttgat ccactcgacc acggcggcgg tgatctcctc 4621 ggcccgcttg tgccggatcg tggcggcgca gaccgggcag agccagatcc gcccgcaccg 4681 catcaggccc aggaccacgg acgttccggc cgccgtctgg gcgacgatca cgccggaggc 4741 agggtccatc agggcgcggc cgcagccctt gcacgcggcg tccccgctga tccgccacag 4801 cgtccggcgg cggctgtacc gggcggcttt ccgcagtcgg gcagcgtcgc tccgcgacgt 4861 gcttcctact tccgagaggc tgtcgcctct cgggctctcc ccatccaccc cgtccggaga 4921 aaccgcaggt cggaggggtg cgggaaactc tgttgtttct ttcccaaggt gttcgctttt 4981 gcctcgggcg gcatctcgcg tcacacgcgc gatcgcccgc ttcgctgcca tccggcagcg 5041 gtctgagcag tagatacgcg gccgtttgcc cggtgtgtgg gcaattgcgg tcccgcagtg 5101 gcagcggggc ccggcgggcc gatctggcaa tgcctcggca tcgctccgta ctctgggcac 5161 gagcaacgtt cctgtctcgc ccggctaagg ggcgcgagtc tgggagcgga cgggtcggag 5221 gtgcgaagtc cggcccgttg ctctttggtc tggtgggaat cctggcacca atcgggccag 5281 aggttccctc cgccactccc gacgcccctt ggggctggtg tgacttggag ggccgaagag 5341 agccccgccg gtgatccggc ggggctttga cgtgcggtca gtgcgtgtgt cggcgagcga 5401 tggccacgag gccctggaag ccgagcggtc cggcgaagtc ggcccagtcg caaccgggct 5461 cagcgcagtg ggcggaccag ccaccgccgt tggggtcctg gaccaggttc acggtcccct 5521 cggtcaggcg tccgtcgaag tcggtcatgg tcggtctcct ggtgggtggg ggcggggcgc 5581 cagcacgaag tgccggcgcc ccgcgggggt tggtcgggtc aggcgccgaa ccggcgggcg 5641 gcggcggcga ccaggccgtc ggcggcggcc atggcgcggt cgcggtcggt ggtgagggcg 5701 gtgcggtcgg cggcggccca gtcctgctct cacgcaacgt ctacgtggac gctcagggcg 5761 acacgatcga ggtcggggag tccgtcgctc cggcgggccg ctggcgcgtc caccgggact 5821 gatcaaggcg aatacttcat atggccctgc aggcggaagt caggtagaca cgacttccgc 5881 tagtccttgc aaggtctgct gacgtgaggc ggggcggtcg tttttgaccg ccctgccttc 5941 gtcatgtagg ctcgctcgct gtgcctggcg tgtcatcaga cgcccaggtc ccggtgccgt 6001 gaggcccggg ccatcgagcc ggtggtacgt ggctgcggtc cccttgtgag ggctgcgcgc 6061 cgtgtgctgt ccggcgcgca cagccttgaa tccacccgcg ggggccggcc ggtctccgtg 6121 agctcgagta gacgacggag acgtaatgtc cgaggtggag ttctcccacg agtactggat 6181 gcgccacgcc ctgaccctgg ccaagcgcgc gtgggacgag cgcgaggtgc cggtgggcgc 6241 cgtgctggtc cacaacaacc gcgtgatcgg cgagggctgg aaccggccga tcggccggca 6301 cgacccgacc gcccacgccg agatcatggc cctgcgccag ggcggcctgg tcatgcagaa 6361 ctaccggctg atcgacgcga ccctgtacgt gaccctggag ccgtgcgtca tgtgcgcggg 6421 cgccatgatc cactcccgca tcggccgggt ggtgttcggc gcccgggacg ccaagaccgg 6481 cgccgcgggc tccctgatgg acgtgctgca ccacccgggc atgaaccacc gcgtcgagat 6541 caccgagggc atcctggcgg acgagtgcgc cgcgctgctg tcggacttct tccggatgcg 6601 ccggcaggag atcaaggccc agaagaaggc gcagtcgtcc accgactcgg gcggctcgtc 6661 cggcggctcg tccggctccg agaccccggg cacctcggag tccgcgaccc cggagtcgtc 6721 cggcggctcg tccggcggct cgtccgaggt cgaattttcc catgaatact ggatgcggca 6781 cgcgctgacc ctggccaagc gggcccgcga cgagcgcgag gtgcccgtcg gcgcggtgct 6841 ggtcctgaac aaccgggtca ttggtgaggg ctggaaccgc gccatcggcc tgcacgaccc 6901 caccgcgcat gccgaaatca tggccctccg gcagggcggc ctggtgatgc agaactaccg 6961 cctcatcgac gccaccctgt acgtgacctt cgagccctgc gtgatgtgtg cgggtgccat 7021 gatccactcc cggatcggcc gcgtcgtgtt cggcgtgcgc aacgccaaga ccggcgcggc 7081 cggctccctg atggacgtcc tgcactaccc cggcatgaac caccgcgtgg aaattaccga 7141 gggcatcctg gccgacgagt gcgcggccct gctgtgctac ttcttccgca tgccgcggca 7201 ggtgttcaac gcgcagaaga aggcgcagtc gtccacggat tcgggcggct cgtccggcgg 7261 ctcgtccggg tcggaaaccc cgggtacctc ggagtcggcc accccggagt cgtccggcgg 7321 ctcgtccggc ggctcggaca agaagtacag catcggcctg gccatcggca ccaacagcgt 7381 gggctgggcg gtcatcaccg acgagtacaa ggtcccctcc aagaagttca aggtcctggg 7441 caacaccgac cggcactcga tcaagaagaa cctgatcggc gccctgctct tcgacagcgg 7501 cgagaccgcc gaggcgaccc gcctgaagcg gaccgcgcgc cggcgctaca cccggcgcaa 7561 gaaccgcatc tgctacctcc aggaaatctt ctccaacgag atggccaagg tcgacgactc 7621 gttcttccac cggctcgagg agagcttcct ggtggaggag gacaagaagc acgagcgcca 7681 cccgatcttc ggcaacatcg tcgacgaggt ggcctaccac gagaagtacc ccaccatcta 7741 ccacctccgc aagaagctgg tcgactcgac cgacaaggcg gacctgcggc tcatctacct 7801 ggccctcgcg cacatgatca agttccgcgg ccacttcctc atcgagggcg acctgaaccc 7861 ggacaactcc gacgtcgaca agctcttcat ccagctggtg cagacctaca accagctgtt 7921 cgaggagaac cccatcaacg ccagcggcgt cgacgccaag gcgatcctct ccgcgcgcct 7981 gagcaagtcc cggcgcctgg agaacctcat cgcccagctg ccgggcgaga agaagaacgg 8041 cctcttcggc aacctgatcg cgctgtcgct cggcctgacc cccaacttca agagcaactt 8101 cgacctggcc gaggacgcga agctccagct gtccaaggac acctacgacg acgacctgga 8161 caacctgctc gcccagatcg gcgaccagta cgcggacctc ttcctggccg cgaagaacct 8221 ctcggacgcc atcctgctca gcgacatcct gcgggtcaac accgagatca ccaaggcccc 8281 gctgtcggcg agcatgatca agcggtacga cgagcaccac caggacctga ccctgctcaa 8341 ggccctcgtg cgccagcagc tgcccgagaa gtacaaggaa atcttcttcg accagtccaa 8401 gaacggctac gccggctaca tcgacggcgg cgcgtcgcag gaggagttct acaagttcat 8461 caagccgatc ctggagaaga tggacggcac cgaggagctg ctcgtcaagc tgaaccgcga 8521 ggacctgctc cgcaagcagc ggaccttcga caacggctcc atcccgcacc agatccacct 8581 gggcgagctg cacgccatcc tccggcgcca ggaggacttc taccccttcc tgaaggacaa 8641 ccgcgagaag atcgagaaga tcctgacctt ccgcatcccg tactacgtcg gccccctggc 8701 ccgcggcaac tcccggttcg cgtggatgac ccggaagtcg gaggagacca tcaccccgtg 8761 gaacttcgag gaggtcgtgg acaagggcgc ctccgcccag tcgttcatcg agcgcatgac 8821 caacttcgac aagaacctcc cgaacgagaa ggtcctgccc aagcactccc tgctctacga 8881 gtacttcacc gtgtacaacg agctgaccaa ggtcaagtac gtgaccgagg gcatgcggaa 8941 gccggccttc ctgtcgggcg agcagaagaa ggcgatcgtc gacctgctct tcaagaccaa 9001 ccgcaaggtc accgtgaagc agctgaagga ggactacttc aagaagatcg agtgcttcga 9061 ctccgtcgag atcagcggcg tggaggaccg cttcaacgcc tccctgggca cctaccacga 9121 cctgctcaag atcatcaagg acaaggactt cctcgacaac gaggagaacg aggacatcct 9181 ggaggacatc gtcctcaccc tgaccctctt cgaggaccgc gagatgatcg aggagcggct 9241 caagacctac gcccacctgt tcgacgacaa ggtgatgaag cagctgaagc ggcgccggta 9301 caccggctgg ggccgcctct cccggaagct gatcaacggc atccgggaca agcagagcgg 9361 caagaccatc ctggacttcc tcaagtccga cggcttcgcc aaccgcaact tcatgcagct 9421 catccacgac gacagcctga ccttcaagga ggacatccag aaggcccagg tctcgggcca 9481 gggcgacagc ctccacgagc acatcgccaa cctggcgggc tccccggcga tcaagaaggg 9541 catcctccag accgtcaagg tcgtggacga gctggtcaag gtgatgggcc gccacaagcc 9601 cgagaacatc gtgatcgaga tggcccggga gaaccagacc acccagaagg gccagaagaa 9661 ctcgcgcgag cggatgaagc ggatcgagga gggcatcaag gagctgggca gccagatcct 9721 gaaggagcac ccggtcgaga acacccagct ccagaacgag aagctgtacc tctactacct 9781 ccagaacggc cgcgacatgt acgtggacca ggagctggac atcaaccggc tgtccgacta 9841 cgacgtcgac gccatcgtgc cgcagtcctt cctgaaggac gactcgatcg acaacaaggt 9901 cctgacccgc tcggacaaga accggggcaa gtccgacaac gtgccctcgg aggaggtcgt 9961 gaagaagatg aagaactact ggcgccagct gctcaacgcc aagctcatca cccagcgcaa 10021 gttcgacaac ctgaccaagg ccgagcgggg cggcctgagc gagctggaca aggcgggctt 10081 catcaagcgc cagctggtcg agacccggca gatcaccaag cacgtggccc agatcctgga 10141 ctcccggatg aacaccaagt acgacgagaa cgacaagctg atccgcgagg tcaaggtgat 10201 caccctcaag agcaagctgg tctccgactt ccgcaaggac ttccagttct acaaggtccg 10261 ggagatcaac aactaccacc acgcccacga cgcgtacctg aacgccgtcg tgggcaccgc 10321 gctgatcaag aagtacccga agctggagtc cgagttcgtc tacggcgact acaaggtcta 10381 cgacgtgcgc aagatgatcg ccaagagcga gcaggagatc ggcaaggcca ccgcgaagta 10441 cttcttctac tccaacatca tgaacttctt caagaccgag atcaccctgg ccaacggcga 10501 gatccgcaag cggcccctga tcgagaccaa cggcgagacc ggcgagatcg tctgggacaa 10561 gggccgcgac ttcgccaccg tccggaaggt gctgtcgatg ccgcaggtca acatcgtgaa 10621 gaagaccgag gtgcagaccg gcggcttcag caaggagtcc atcctcccca agcgcaacag 10681 cgacaagctg atcgcccgga agaaggactg ggacccgaag aagtacggcg gcttcgacag 10741 ccccaccgtc gcctactccg tgctggtcgt ggcgaaggtc gagaagggca agagcaagaa 10801 gctgaagtcc gtgaaggagc tgctcggcat caccatcatg gagcgctcct cgttcgagaa 10861 gaacccgatc gacttcctgg aggccaaggg ctacaaggag gtcaagaagg acctcatcat 10921 caagctgccc aagtacagcc tgttcgagct ggagaacggc cgcaagcgga tgctcgcctc 10981 cgcgggcgag ctgcaaaagg gcaacgagct ggccctcccg tcgaagtacg tcaacttcct 11041 gtacctcgcg tcccactacg agaagctgaa gggctcgccc gaggacaacg agcagaagca 11101 gctcttcgtg gagcagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt 11161 cagcaagcgc gtcatcctgg ccgacgcgaa cctcgacaag gtgctgtccg cctacaacaa 11221 gcaccgcgac aagccgatcc gggagcaggc ggagaacatc atccacctgt tcaccctcac 11281 caacctgggt gccccggccg ccttcaagta cttcgacacc accatcgacc gcaagcggta 11341 cacctccacc aaggaggtcc tcgacgcgac cctgatccac cagagcatca ccggcctgta 11401 cgagacccgc atcgacctga gccagctggg cggcgactga gaattcagat ctacgcgttc 11461 cccgcaaaag cggcctttga ctccctgcaa gcctcagcga ccgaatatat cggttatgcg 11521 tgggcgatgg ttgttgtcat tgtcggcgca actatcggta tcaagctgtt taagaaattc 11581 acctcgaaag caagctgata aaccgataca attaaaggct ccttttggag ccttttttta 11641 agcttgcatg cctgcaggtc gacggatctt ttccgctgca taaccctgct tcggggtcat 11701 tatagcgatt ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag 11761 ggttcgtgta gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt 11821 aggcccaccc gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct 11881 caacgggaat cctgctctgc gaggctggcc ggctaccgcc ggcgtaacag atgagggcaa 11941 gcggatggct gatgaaacca agccaaccag gaagggcagc ccacctatca aggtgtactg 12001 ccttccagac gaacgaagag cgattgagga aaaggcggcg gcggccggca tgagcctgtc 12061 ggcctacctg ctggccgtcg gccagggcta caaaatcacg ggcgtcgtgg actatgagca 12121 cgtccgcgag ctggcccgca tcaatggcga cctgggccgc ctgggcggcc tgctgaaact 12181 ctggctcacc gacgacccgc gcacggcgcg gttcggtgat gccacgatcc tcgccctgct 12241 ggcgaagatc gaagagaagc aggacgagct tggcaaggtc atgatgggcg tggtccgccc 12301 gagggcagag ccatgacttt tttagccgct aaaacggccg gggggtgcgc gtgattgcca 12361 agcacgtccc catgcgctcc atcaagaaga gcgacttcgc ggagctggtg aagtacatca 12421 ccgacgagca aggcaagacc gatccccggg gacctgcagg tcgactctag agttttagag 12481 ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag 12541 tcggtgctaa aaaaaaaccc cgcccctgac agggcggggt tttttttgta gcctcaggag 12601 ttgtagctgt tcacattcga acggtctctg ctttgacaac atgctgtgcg gtgttgtaaa 12661 gtcgtggccg ctagctattt acccgcagga catatccacg ccctcctaca tcgaagctga 12721 aagcacgaga ttcttcgccc tccgagagct gcatcaggtc ggagacgctg tcgaactttt 12781 cgatcagaaa cttctcgaca gacgtcgcgg tgagttcagg ctttttcata tctcattgcc 12841 cccggacgag cgtctgctcc gccattcgcc gtccgccgtg ccaatcggat cagccgtcca 12901 aatgcgggat tttcgttagt cggaggccaa acggcattga gcgtcagcat atcatcagcg 12961 agctgaagaa agacaatccc cgatccgctc cacgtgttgc cccagcaatc agcgcgacct 13021 tgcccctcca acgtcatctc gttctccgct catgagctca gccaatcgac tggcgagcgg 13081 catcgcattc ttcgcatccc gcctctggcg gatgcaggaa gatcaacgga tctcggccca 13141 gttgacccag ggctgtcgcc acaatgtcgc gggagcggat caaccgagca aaggcatgac 13201 cgactggacc ttccttctga aggctcttct ccttgagcca cctgtccgcc aaggcaaagc 13261 gctcacagca gtggtcattc tcgagataat cgacgcgtac caacttgcca tcctgaagaa 13321 tggtgcagtg tctcggcacc ccatagggaa cctttgccat caactcggca agatgcagcg 13381 tcgtgttggc atcgtgtccc acgccgagga gaagtacctg cccatcgagt tcatggacac 13441 gggcgaccgg gcttgcaggc gagtgaggtg gcaggggcaa tggatcagag atgatctgct 13501 ctgcctgtgg ccccgctgcc gcaaaggcaa atggatgggc gctgcgcttt acatttggca 13561 ggcgccagaa tgtgtcagag acaactccaa ggtccggtgt aacgggcgac gtggcaggat 13621 cgaacggctc gtcgtccaga cctgaccacg agggcatgac gagcgtccct cccggaccca 13681 gcgcagcacg cagggcctcg atcagtccaa gtggcccatc ttcgaggggc cggacgctac 13741 ggaaggagct gtggaccagc agcacaccgc cgggggtaac cccaaggttg agaagctgac 13801 cgatg //