LOCUS Exported 13610 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses BE3 CRISPR base editor and gRNA scaffold in Streptomyces. ACCESSION . VERSION . KEYWORDS pSCBE3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13610) 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 13610) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13610 /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 6146..6832 /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 6881..10981 /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 10994..11245 /codon_start=1 /product="uracil-DNA glycosylase inhibitor from a Bacillus subtilis bacteriophage (Mol et al., 1995)" /label=UGI /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML" oriT 11608..11717 /note="incP origin of transfer" CDS 11750..12121 /codon_start=1 /gene="traJ" /product="oriT-recognizing protein" /label=traJ /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE KQDELGKVMMGVVRPRAEP" misc_RNA 12277..12352 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" CDS complement(12866..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 gtccgagacc ggcccggtgg ccgtggaccc 6181 gaccctgcgt cgccgcatcg agccgcacga gttcgaggtg ttcttcgacc cgcgggagct 6241 gcgcaaggag acctgcctgc tgtacgagat caactggggc ggccggcact cgatctggcg 6301 ccacacctcg cagaacacca acaagcacgt cgaggtgaac ttcatcgaga agttcaccac 6361 cgagcggtac ttctgcccga acacccgctg ctcgatcacc tggttcctgt cctggtcccc 6421 gtgcggcgag tgctcccggg cgatcaccga gttcctgtcc cgctacccgc acgtcaccct 6481 gttcatctac atcgcccggc tgtaccacca cgccgacccg cggaaccggc agggcctgcg 6541 cgacctgatc tcctccggcg tgaccatcca gatcatgacc gagcaggagt ccggctactg 6601 ctggcggaac ttcgtcaact actcgccgtc caacgaggcc cactggccgc ggtacccgca 6661 cctgtgggtg cgcctgtacg tcctggagct gtactgcatc atcctgggcc tgccgccgtg 6721 cctgaacatc ctgcgtcgca agcagccgca gctgaccttc ttcaccatcg ccctgcagtc 6781 ctgccactac cagcgcctgc cgccgcacat cctgtgggcg accggcctga agtccggctc 6841 ggagaccccg ggcacctcgg agtccgccac cccggagtcc gacaagaagt acagcatcgg 6901 cctggccatc ggcaccaaca gcgtgggctg ggcggtcatc accgacgagt acaaggtccc 6961 ctccaagaag ttcaaggtcc tgggcaacac cgaccggcac tcgatcaaga agaacctgat 7021 cggcgccctg ctcttcgaca gcggcgagac cgccgaggcg acccgcctga agcggaccgc 7081 gcgccggcgc tacacccggc gcaagaaccg catctgctac ctccaggaaa tcttctccaa 7141 cgagatggcc aaggtcgacg actcgttctt ccaccggctc gaggagagct tcctggtgga 7201 ggaggacaag aagcacgagc gccacccgat cttcggcaac atcgtcgacg aggtggccta 7261 ccacgagaag taccccacca tctaccacct ccgcaagaag ctggtcgact cgaccgacaa 7321 ggcggacctg cggctcatct acctggccct cgcgcacatg atcaagttcc gcggccactt 7381 cctcatcgag ggcgacctga acccggacaa ctccgacgtc gacaagctct tcatccagct 7441 ggtgcagacc tacaaccagc tgttcgagga gaaccccatc aacgccagcg gcgtcgacgc 7501 caaggcgatc ctctccgcgc gcctgagcaa gtcccggcgc ctggagaacc tcatcgccca 7561 gctgccgggc gagaagaaga acggcctctt cggcaacctg atcgcgctgt cgctcggcct 7621 gacccccaac ttcaagagca acttcgacct ggccgaggac gcgaagctcc agctgtccaa 7681 ggacacctac gacgacgacc tggacaacct gctcgcccag atcggcgacc agtacgcgga 7741 cctcttcctg gccgcgaaga acctctcgga cgccatcctg ctcagcgaca tcctgcgggt 7801 caacaccgag atcaccaagg ccccgctgtc ggcgagcatg atcaagcggt acgacgagca 7861 ccaccaggac ctgaccctgc tcaaggccct cgtgcgccag cagctgcccg agaagtacaa 7921 ggaaatcttc ttcgaccagt ccaagaacgg ctacgccggc tacatcgacg gcggcgcgtc 7981 gcaggaggag ttctacaagt tcatcaagcc gatcctggag aagatggacg gcaccgagga 8041 gctgctcgtc aagctgaacc gcgaggacct gctccgcaag cagcggacct tcgacaacgg 8101 ctccatcccg caccagatcc acctgggcga gctgcacgcc atcctccggc gccaggagga 8161 cttctacccc ttcctgaagg acaaccgcga gaagatcgag aagatcctga ccttccgcat 8221 cccgtactac gtcggccccc tggcccgcgg caactcccgg ttcgcgtgga tgacccggaa 8281 gtcggaggag accatcaccc cgtggaactt cgaggaggtc gtggacaagg gcgcctccgc 8341 ccagtcgttc atcgagcgca tgaccaactt cgacaagaac ctcccgaacg agaaggtcct 8401 gcccaagcac tccctgctct acgagtactt caccgtgtac aacgagctga ccaaggtcaa 8461 gtacgtgacc gagggcatgc ggaagccggc cttcctgtcg ggcgagcaga agaaggcgat 8521 cgtcgacctg ctcttcaaga ccaaccgcaa ggtcaccgtg aagcagctga aggaggacta 8581 cttcaagaag atcgagtgct tcgactccgt cgagatcagc ggcgtggagg accgcttcaa 8641 cgcctccctg ggcacctacc acgacctgct caagatcatc aaggacaagg acttcctcga 8701 caacgaggag aacgaggaca tcctggagga catcgtcctc accctgaccc tcttcgagga 8761 ccgcgagatg atcgaggagc ggctcaagac ctacgcccac ctgttcgacg acaaggtgat 8821 gaagcagctg aagcggcgcc ggtacaccgg ctggggccgc ctctcccgga agctgatcaa 8881 cggcatccgg gacaagcaga gcggcaagac catcctggac ttcctcaagt ccgacggctt 8941 cgccaaccgc aacttcatgc agctcatcca cgacgacagc ctgaccttca aggaggacat 9001 ccagaaggcc caggtctcgg gccagggcga cagcctccac gagcacatcg ccaacctggc 9061 gggctccccg gcgatcaaga agggcatcct ccagaccgtc aaggtcgtgg acgagctggt 9121 caaggtgatg ggccgccaca agcccgagaa catcgtgatc gagatggccc gggagaacca 9181 gaccacccag aagggccaga agaactcgcg cgagcggatg aagcggatcg aggagggcat 9241 caaggagctg ggcagccaga tcctgaagga gcacccggtc gagaacaccc agctccagaa 9301 cgagaagctg tacctctact acctccagaa cggccgcgac atgtacgtgg accaggagct 9361 ggacatcaac cggctgtccg actacgacgt cgaccacatc gtgccgcagt ccttcctgaa 9421 ggacgactcg atcgacaaca aggtcctgac ccgctcggac aagaaccggg gcaagtccga 9481 caacgtgccc tcggaggagg tcgtgaagaa gatgaagaac tactggcgcc agctgctcaa 9541 cgccaagctc atcacccagc gcaagttcga caacctgacc aaggccgagc ggggcggcct 9601 gagcgagctg gacaaggcgg gcttcatcaa gcgccagctg gtcgagaccc ggcagatcac 9661 caagcacgtg gcccagatcc tggactcccg gatgaacacc aagtacgacg agaacgacaa 9721 gctgatccgc gaggtcaagg tgatcaccct caagagcaag ctggtctccg acttccgcaa 9781 ggacttccag ttctacaagg tccgggagat caacaactac caccacgccc acgacgcgta 9841 cctgaacgcc gtcgtgggca ccgcgctgat caagaagtac ccgaagctgg agtccgagtt 9901 cgtctacggc gactacaagg tctacgacgt gcgcaagatg atcgccaaga gcgagcagga 9961 gatcggcaag gccaccgcga agtacttctt ctactccaac atcatgaact tcttcaagac 10021 cgagatcacc ctggccaacg gcgagatccg caagcggccc ctgatcgaga ccaacggcga 10081 gaccggcgag atcgtctggg acaagggccg cgacttcgcc accgtccgga aggtgctgtc 10141 gatgccgcag gtcaacatcg tgaagaagac cgaggtgcag accggcggct tcagcaagga 10201 gtccatcctc cccaagcgca acagcgacaa gctgatcgcc cggaagaagg actgggaccc 10261 gaagaagtac ggcggcttcg acagccccac cgtcgcctac tccgtgctgg tcgtggcgaa 10321 ggtcgagaag ggcaagagca agaagctgaa gtccgtgaag gagctgctcg gcatcaccat 10381 catggagcgc tcctcgttcg agaagaaccc gatcgacttc ctggaggcca agggctacaa 10441 ggaggtcaag aaggacctca tcatcaagct gcccaagtac agcctgttcg agctggagaa 10501 cggccgcaag cggatgctcg cctccgcggg cgagctgcaa aagggcaacg agctggccct 10561 cccgtcgaag tacgtcaact tcctgtacct cgcgtcccac tacgagaagc tgaagggctc 10621 gcccgaggac aacgagcaga agcagctctt cgtggagcag cacaagcact acctggacga 10681 gatcatcgag cagatcagcg agttcagcaa gcgcgtcatc ctggccgacg cgaacctcga 10741 caaggtgctg tccgcctaca acaagcaccg cgacaagccg atccgggagc aggcggagaa 10801 catcatccac ctgttcaccc tcaccaacct gggtgccccg gccgccttca agtacttcga 10861 caccaccatc gaccgcaagc ggtacacctc caccaaggag gtcctcgacg cgaccctgat 10921 ccaccagagc atcaccggcc tgtacgagac ccgcatcgac ctgagccagc tgggcggcga 10981 ctcgggcggc tccaccaacc tgtcggacat catcgagaag gagaccggca agcagctggt 11041 gatccaggag tccatcctga tgctgccgga ggaagtggag gaagtgatcg gcaacaagcc 11101 ggagtcggac atcctggtgc acaccgccta cgacgagtcg accgacgaga acgtcatgct 11161 gctgacctcg gacgccccgg agtacaagcc gtgggccctg gtcatccagg actccaacgg 11221 cgagaacaag atcaagatgc tgtgagaatt cagatctacg cgttccccgc aaaagcggcc 11281 tttgactccc tgcaagcctc agcgaccgaa tatatcggtt atgcgtgggc gatggttgtt 11341 gtcattgtcg gcgcaactat cggtatcaag ctgtttaaga aattcacctc gaaagcaagc 11401 tgataaaccg atacaattaa aggctccttt tggagccttt ttttaagctt gcatgcctgc 11461 aggtcgacgg atcttttccg ctgcataacc ctgcttcggg gtcattatag cgattttttc 11521 ggtatatcca tcctttttcg cacgatatac aggattttgc caaagggttc gtgtagactt 11581 tccttggtgt atccaacggc gtcagccggg caggataggt gaagtaggcc cacccgcgag 11641 cgggtgttcc ttcttcactg tcccttattc gcacctggcg gtgctcaacg ggaatcctgc 11701 tctgcgaggc tggccggcta ccgccggcgt aacagatgag ggcaagcgga tggctgatga 11761 aaccaagcca accaggaagg gcagcccacc tatcaaggtg tactgccttc cagacgaacg 11821 aagagcgatt gaggaaaagg cggcggcggc cggcatgagc ctgtcggcct acctgctggc 11881 cgtcggccag ggctacaaaa tcacgggcgt cgtggactat gagcacgtcc gcgagctggc 11941 ccgcatcaat ggcgacctgg gccgcctggg cggcctgctg aaactctggc tcaccgacga 12001 cccgcgcacg gcgcggttcg gtgatgccac gatcctcgcc ctgctggcga agatcgaaga 12061 gaagcaggac gagcttggca aggtcatgat gggcgtggtc cgcccgaggg cagagccatg 12121 acttttttag ccgctaaaac ggccgggggg tgcgcgtgat tgccaagcac gtccccatgc 12181 gctccatcaa gaagagcgac ttcgcggagc tggtgaagta catcaccgac gagcaaggca 12241 agaccgatcc ccggggacct gcaggtcgac tctagagttt tagagctaga aatagcaagt 12301 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctaaaaaaa 12361 aaccccgccc ctgacagggc ggggtttttt ttgtagcctc aggagttgta gctgttcaca 12421 ttcgaacggt ctctgctttg acaacatgct gtgcggtgtt gtaaagtcgt ggccgctagc 12481 tatttacccg caggacatat ccacgccctc ctacatcgaa gctgaaagca cgagattctt 12541 cgccctccga gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc agaaacttct 12601 cgacagacgt cgcggtgagt tcaggctttt tcatatctca ttgcccccgg acgagcgtct 12661 gctccgccat tcgccgtccg ccgtgccaat cggatcagcc gtccaaatgc gggattttcg 12721 ttagtcggag gccaaacggc attgagcgtc agcatatcat cagcgagctg aagaaagaca 12781 atccccgatc cgctccacgt gttgccccag caatcagcgc gaccttgccc ctccaacgtc 12841 atctcgttct ccgctcatga gctcagccaa tcgactggcg agcggcatcg cattcttcgc 12901 atcccgcctc tggcggatgc aggaagatca acggatctcg gcccagttga cccagggctg 12961 tcgccacaat gtcgcgggag cggatcaacc gagcaaaggc atgaccgact ggaccttcct 13021 tctgaaggct cttctccttg agccacctgt ccgccaaggc aaagcgctca cagcagtggt 13081 cattctcgag ataatcgacg cgtaccaact tgccatcctg aagaatggtg cagtgtctcg 13141 gcaccccata gggaaccttt gccatcaact cggcaagatg cagcgtcgtg ttggcatcgt 13201 gtcccacgcc gaggagaagt acctgcccat cgagttcatg gacacgggcg accgggcttg 13261 caggcgagtg aggtggcagg ggcaatggat cagagatgat ctgctctgcc tgtggccccg 13321 ctgccgcaaa ggcaaatgga tgggcgctgc gctttacatt tggcaggcgc cagaatgtgt 13381 cagagacaac tccaaggtcc ggtgtaacgg gcgacgtggc aggatcgaac ggctcgtcgt 13441 ccagacctga ccacgagggc atgacgagcg tccctcccgg acccagcgca gcacgcaggg 13501 cctcgatcag tccaagtggc ccatcttcga ggggccggac gctacggaag gagctgtgga 13561 ccagcagcac accgccgggg gtaaccccaa ggttgagaag ctgaccgatg //