LOCUS Exported 12247 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION C to T base editor for actinomycetes. ACCESSION . VERSION . KEYWORDS pCRISPR-cBEST SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12247) AUTHORS Tong Y, Whitford CM, Robertsen HL, Blin K, Jorgensen TS, Klitgaard AK, Gren T, Jiang X, Weber T, Lee SY TITLE Highly efficient DSB-free base editing for streptomycetes with CRISPR-BEST. JOURNAL Proc Natl Acad Sci U S A. 2019 Oct 8;116(41):20366-20375. doi: 10.1073/pnas.1913493116. Epub 2019 Sep 23. PUBMED 31548381 REFERENCE 2 (bases 1 to 12247) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12247 /organism="synthetic DNA construct" /mol_type="other DNA" oriT 466..575 /note="incP origin of transfer" CDS 891..1691 /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 LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDSFWRLPNV KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH LAELLAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG" rep_origin 1993..2580 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2481..2500 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2734..2751 /label=L4440 /note="L4440 vector, forward primer" protein_bind 2868..2889 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2904..2934 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2942..2958 /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 2947..2969 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2966..2982 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2966..2982 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" misc_RNA 3119..3194 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" terminator 3364..3398 /label=lambda t0 terminator /note="minimal transcription terminator from phage lambda (Scholtissek and Grosse, 1987)" CDS 3566..4252 /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 4301..8401 /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 8414..8665 /codon_start=1 /product="uracil-DNA glycosylase inhibitor from a Bacillus subtilis bacteriophage (Mol et al., 1995)" /label=UGI /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML" terminator 8846..8894 /label=fd terminator /note="central terminator from bacteriophage fd (Otsuka and Kunisawa, 1982)" CDS 10609..12051 /codon_start=1 /gene="rep" /product="replication initiator protein from the Streptomyces ghanaensis plasmid pSG5" /label=pSG5 Rep /translation="MWKPGEATWGNTCRCNNVHTCPWCMSRILAVRGSNVQLAADGLAD AGYGLHLGTNTLRHFERMAFGTVRKGMRHGLVAVLHDGWKGAYGSSGRRWRTMRDDFGI IGYERAFEDTFGWGSGWHLHWHTLWVTREVLGPDAQAAFRDALAGAWAAGVESAGGYTV SETCDRPGCSCEGKGHGTDVRPLNGADAADGDAGKQARYLYKDGDKTKGGVAKIGLELA GQNFKAGRGDDRMGPLDLGDAAAAELQRLRRPGPFVEKYREREFGVFQVRKHYRSQNLN RLIKELGIQQDVRTEEEITDDTEGLVAIAVIPAYIWYRYIARVAGRRLDLIKVAETYGL PGVRRLVESWGLVWGKDVLDPPAPEAPAAPGDLDADQMRFEVMSEEEAAFREARRKANE ARTEELAASLDRVRQPKKEAIRPTISLRKRLKPKPVTVDVKTPPPGAASPVCRRCKGKL APVLQPWGQHPGDCLRVDTAVA" ORIGIN 1 ggccgtcgac gacgacgcgt ccgcctgcct gcttttggcc ggcgggccgg gccgcggttg 61 tccgccgggt cggaccggtc ccttggtcga ccgatgcctc ggccacgacg atgctgggcc 121 cgaagcggta cgcgacccgt gccgggccgc agcggatcgc ggcagcagga gctccccgcc 181 cgccacggct ggctggatgg tcgcctcacg cgcccgcagg ggtgacggcg tgggtgcggc 241 ggtcagcccc gtgatcgggc gtcagacggc cgctgaggga ccgcgaccac ccctcccttc 301 ccgcctgccg tctttcaccg cccggccggt ccgcagacgc cgtacggcca ggccacggag 361 gattacgggc agatggcgcc cacccggcac ccgggctggg gcaccctgga cagcacgaag 421 gtccgcccca ctcaccgggg cgggccttcg cgttgagcta gagttcccgc cagcctcgca 481 gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc 541 cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg atacaccaag 601 gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata 661 ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaaaat gcagctcacg 721 gtaactgatg ccgtatttgc agtaccagcg tacggcccac agaatgatgt cacgctgaaa 781 atgccggcct ttgaatgggt tcatgtgcag ctccataagc aaaaggggat gataagttta 841 tcaccaccga ctatttgcaa cagtgccgtt gatcgtgcta tgatcgactg atgtcatcag 901 cggtggagtg caatgtcgtg caatacgaat ggcgaaaagc cgagctcatc ggtcagcttc 961 tcaaccttgg ggttaccccc ggcggtgtgc tgctggtcca cagctccttc cgtagcgtcc 1021 ggcccctcga agatgggcca cttggactga tcgaggccct gcgtgctgcg ctgggtccgg 1081 gagggacgct cgtcatgccc tcgtggtcag gtctggacga cgagccgttc gatcctgcca 1141 cgtcgcccgt tacaccggac cttggagttg tctctgactc attctggcgc ctgccaaatg 1201 taaagcgcag cgcccatcca tttgcctttg cggcagcggg gccacaggca gagcagatca 1261 tctctgatcc attgcccctg ccacctcact cgcctgcaag cccggtcgcc cgtgtccatg 1321 aactcgatgg gcaggtactt ctcctcggcg tgggacacga tgccaacacg acgctgcatc 1381 ttgccgagtt gttggcaaag gttccctatg gggtgccgag acactgcacc attcttcagg 1441 atggcaagtt ggtacgcgtc gattatctcg agaatgacca ctgctgtgag cgctttgcct 1501 tggcggacag gtggctcaag gagaagagcc ttcagaagga aggtccagtc ggtcatgcct 1561 ttgctcggtt gatccgctcc cgcgacattg tggcgacagc cctgggtcaa ctgggccgag 1621 atccgttgat cttcctgcat ccgccagagg cgggatgcga agaatgcgat gccgctcgcc 1681 agtcgattgg ctgagctcat aagttcctat tccgaagttc ctattcgaaa tgaccgacca 1741 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt 1801 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat 1861 gctggagttc ttcgcccacc ccaaaaggat ctaggtgaag atcctttttg ataatctcat 1921 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat 1981 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa 2041 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa 2101 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt 2161 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt 2221 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata 2281 gttaccggat aaggcgcagc ggtcgggctg aacggggggt cgtgcacaca gcccagcttg 2341 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 2401 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 2461 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 2521 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 2581 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 2641 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct 2701 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa 2761 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg 2821 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag 2881 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga 2941 attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac gaattgtacg 3001 cggtcgatct tgacggctgg cgagaggtgc ggggaggatc tgaccgacgc ggtccacacg 3061 tggcaccgcg atgctgttgt gggcacaatc gtgccggttg gtaggatcga cggccatggt 3121 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg 3181 caccgagtcg gtgctttttt tacgtaggcg tcagctacga tgttccgggg actgctgatc 3241 cggtcagcag gtggaagagg gactggattc caaagttctc aatgctgctt gctgttcttg 3301 aatggggggt cgttgacgac gacatggctc gattggcgcg acaagttgct gcgattctca 3361 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag 3421 gtcattactg gaccggatcg gggatctggg ctgagggagc cgacggcacg cggcggctca 3481 cggcgtggca cgcggaacgt ccgggcttgc acctcacgtc acgtgaggag gcagcgtgga 3541 cggcgtcaga gaagggagcg gacatatgtc ctccgagacc ggccccgtgg ccgtggaccc 3601 gaccctgcgt cgccgcatcg agccgcacga gttcgaggtc ttcttcgacc cgcgcgagct 3661 gcgcaaggag acctgcctgc tgtacgagat caactggggc ggccggcact ccatctggcg 3721 ccacacctcg cagaacacca acaagcacgt cgaggtcaac ttcatcgaga agttcaccac 3781 cgagcggtac ttctgcccga acacccgctg ctccatcacc tggttcctgt cctggtcccc 3841 ctgcggcgag tgctcccggg ccatcaccga gttcctgtcc cgctacccgc acgtcaccct 3901 gttcatctac atcgcccggc tgtaccacca cgccgacccg cgcaaccggc agggcctgcg 3961 cgacctgatc tcctccggcg tcaccatcca gatcatgacc gagcaggagt ccggctactg 4021 ctggcggaac ttcgtcaact actccccgtc caacgaggcc cactggcccc gctacccgca 4081 cctgtgggtc cgcctgtacg tcctggagct gtactgcatc atcctgggcc tgccgccctg 4141 cctgaacatc ctgcgtcgca agcagccgca gctgaccttc ttcaccatcg ccctccagtc 4201 ctgccactac cagcgcctgc cgccccacat cctgtgggcc accggcctga agtccggctc 4261 cgagaccccg ggcacctcgg agtccgccac cccggagtcc gacaagaagt actccatcgg 4321 cctcgcgatc ggcaccaact ccgtgggctg ggcggtcatc accgacgagt acaaggtccc 4381 ctccaagaag ttcaaggtcc tgggcaacac cgaccggcac tcgatcaaga agaacctgat 4441 cggcgccctg ctcttcgaca gcggcgagac cgccgaggcg acccgcctga agcggaccgc 4501 gcgtcgccgc tacacccggc gcaagaaccg catctgctac ctgcaggaaa tcttctccaa 4561 cgagatggcc aaggtggacg actcgttctt ccaccgcctg gaggagagct tcctggtgga 4621 ggaggacaag aagcacgagc gccacccgat cttcggcaac atcgtggacg aggtggccta 4681 ccacgagaag taccccacca tctaccacct ccgcaagaag ctggtggact cgaccgacaa 4741 ggcggacctg cggctcatct acctggccct cgcgcacatg atcaagttcc gcggccactt 4801 cctcatcgag ggcgacctga acccggacaa ctccgacgtg gacaagctct tcatccagct 4861 ggtgcagacc tacaaccagc tgttcgagga gaaccccatc aacgccagcg gcgtggacgc 4921 caaggcgatc ctctccgcgc gcctgagcaa gtcccggcgc ctggagaacc tcatcgccca 4981 gctgccgggc gagaagaaga acggcctctt cggcaacctg atcgcgctgt cgctcggcct 5041 gacccccaac ttcaagagca acttcgacct ggccgaggac gcgaagctcc agctgtccaa 5101 ggacacctac gacgacgacc tggacaacct gctcgcccag atcggcgacc agtacgcgga 5161 cctcttcctg gccgcgaaga acctctcgga cgccatcctg ctcagcgaca tcctgcgggt 5221 caacaccgag atcaccaagg ccccgctgtc ggcgagcatg atcaagcggt acgacgagca 5281 ccaccaggac ctgaccctgc tcaaggccct cgtgcgccag cagctgcccg agaagtacaa 5341 ggaaatcttc ttcgaccagt ccaagaacgg ctacgccggc tacatcgacg gcggcgcgtc 5401 gcaggaggag ttctacaagt tcatcaagcc gatcctggag aagatggacg gcaccgagga 5461 gctgctcgtc aagctgaacc gcgaggacct gctccgcaag cagcggacct tcgacaacgg 5521 ctccatcccg caccagatcc acctgggcga gctccacgcc atcctccggc gccaggagga 5581 cttctacccc ttcctgaagg acaaccgcga gaagatcgag aagatcctga ccttccgcat 5641 cccgtactac gtcggccccc tggcccgcgg caactcccgg ttcgcgtgga tgacccggaa 5701 gtcggaggag accatcaccc cgtggaactt cgaggaggtc gtggacaagg gcgcgtccgc 5761 gcagtcgttc atcgagcgca tgaccaactt cgacaagaac ctcccgaacg agaaggtcct 5821 gcccaagcac tccctgctct acgagtactt caccgtgtac aacgagctga ccaaggtcaa 5881 gtacgtgacc gagggcatgc ggaagccggc cttcctgtcg ggcgagcaga agaaggcgat 5941 cgtggacctg ctcttcaaga ccaaccgcaa ggtcaccgtg aagcagctga aggaggacta 6001 cttcaagaag atcgagtgct tcgactccgt cgagatcagc ggcgtggagg accgcttcaa 6061 cgcctccctg ggcacctacc acgacctgct caagatcatc aaggacaagg acttcctcga 6121 caacgaggag aacgaggaca tcctggagga catcgtcctc accctgaccc tcttcgagga 6181 ccgcgagatg atcgaggagc ggctcaagac ctacgcccac ctgttcgacg acaaggtgat 6241 gaagcagctg aagcgtcgcc gctacaccgg ctggggccgc ctctcccgga agctgatcaa 6301 cggcatccgg gacaagcaga gcggcaagac catcctggac ttcctcaagt ccgacggctt 6361 cgccaaccgc aacttcatgc agctcatcca cgacgacagc ctgaccttca aggaggacat 6421 ccagaaggcc caggtctcgg gccagggcga cagcctccac gagcacatcg ccaacctggc 6481 gggctccccg gcgatcaaga agggcatcct ccagaccgtc aaggtcgtgg acgagctggt 6541 caaggtgatg ggccgccaca agcccgagaa catcgtgatc gagatggccc gggagaacca 6601 gaccacccag aagggccaga agaactcgcg cgagcggatg aagcggatcg aggagggcat 6661 caaggagctc ggcagccaga tcctgaagga gcacccggtc gagaacaccc agctgcagaa 6721 cgagaagctg tacctctact acctgcagaa cggccgcgac atgtacgtgg accaggagct 6781 cgacatcaac cggctgtccg actacgacgt ggaccacatc gtgccgcagt ccttcctgaa 6841 ggacgactcg atcgacaaca aggtcctgac ccgctcggac aagaaccggg gcaagtccga 6901 caacgtgccc tcggaggagg tcgtgaagaa gatgaagaac tactggcgcc agctgctcaa 6961 cgccaagctc atcacccagc gcaagttcga caacctgacc aaggccgagc ggggcggcct 7021 gagcgagctc gacaaggcgg gcttcatcaa gcgccagctg gtcgagaccc ggcagatcac 7081 caagcacgtg gcccagatcc tggactcccg gatgaacacc aagtacgacg agaacgacaa 7141 gctgatccgc gaggtcaagg tgatcaccct caagagcaag ctggtctccg acttccgcaa 7201 ggacttccag ttctacaagg tccgggagat caacaactac caccacgccc acgacgcgta 7261 cctgaacgcc gtcgtgggca ccgcgctgat caagaagtac ccgaagctgg agtccgagtt 7321 cgtctacggc gactacaagg tctacgacgt gcgcaagatg atcgccaaga gcgagcagga 7381 gatcggcaag gccaccgcga agtacttctt ctactccaac atcatgaact tcttcaagac 7441 cgagatcacc ctggccaacg gcgagatccg caagcggccc ctgatcgaga ccaacggcga 7501 gaccggcgag atcgtctggg acaagggccg cgacttcgcc accgtccgga aggtgctgtc 7561 gatgccgcag gtcaacatcg tgaagaagac cgaggtgcag accggcggct tcagcaagga 7621 gtccatcctc cccaagcgca acagcgacaa gctgatcgcc cggaagaagg actgggaccc 7681 gaagaagtac ggcggcttcg acagccccac cgtcgcctac tccgtgctgg tcgtggcgaa 7741 ggtcgagaag ggcaagagca agaagctgaa gtccgtgaag gagctgctcg gcatcaccat 7801 catggagcgc tcctcgttcg agaagaaccc gatcgacttc ctggaggcca agggctacaa 7861 ggaggtcaag aaggacctca tcatcaagct gcccaagtac agcctgttcg agctggagaa 7921 cggccgcaag cggatgctcg cctccgcggg cgagctgcag aagggcaacg agctggccct 7981 cccgtcgaag tacgtcaact tcctgtacct cgcgtcccac tacgagaagc tgaagggctc 8041 gcccgaggac aacgagcaga agcagctctt cgtggagcag cacaagcact acctggacga 8101 gatcatcgag cagatcagcg agttcagcaa gcgcgtcatc ctggccgacg cgaacctcga 8161 caaggtgctg tccgcctaca acaagcaccg cgacaagccg atccgggagc aggcggagaa 8221 catcatccac ctgttcaccc tcaccaacct gggcgccccc gccgcgttca agtacttcga 8281 caccaccatc gaccgcaagc ggtacacctc caccaaggag gtcctcgacg cgaccctgat 8341 ccaccagagc atcaccggcc tgtacgagac ccgcatcgac ctgtcccagc tcggcggcga 8401 ctccggcggc tccaccaacc tgtccgacat catcgagaag gagaccggca agcagctggt 8461 catccaggag tccatcctga tgctgccgga ggaagtcgag gaagtcatcg gcaacaagcc 8521 cgagtccgac atcctggtcc acaccgccta cgacgagtcc accgacgaga acgtcatgct 8581 gctgacctcg gacgcccccg agtacaagcc ctgggccctg gtcatccagg actccaacgg 8641 cgagaacaag atcaagatgc tgtgatctag aggatccccg ggtaccgagc tcgaattccc 8701 cagatctaaa gttttgtcgt ctttccagac gttagtaaat gaattttctg tatgaggttt 8761 tgctaaacaa ctttcaacag tttcagcgga gtgagaatag aaaggaacaa ctaaaggaat 8821 tgcgaataat aattttttca cgttgaaaat ctccaaaaaa aaaggctcca aaaggagcct 8881 ttaattgtat cggtttatca gcttgctttc gaggtgaatt tcttaaacag cttgataccg 8941 atagttgcgc cgacaatgac aacaaccatc gcccacgcat aaccgatata ttcggtcgct 9001 gaggcttgca gggagtcaaa ggccgctttt gcgggatctc gtcgaaggca ctagaggcct 9061 atatgcgggg atcgaccgcg cgggtcccgg acggggaaga gcggggagct ttgccagaga 9121 gcgacgactt ccccttgcgt tggtgattgc cggtcagggc agccatccgc catcgtcgcg 9181 tagggtgtca caccccagga atcgcgtcac tgaacacagc agccggtagg acgaccatga 9241 ctgagttgga caccatcgca aatccgtccg atcccgcggt gcagcggatc atcgatgtca 9301 ccaagccgtc gcgatccaac ataaagacaa cgttgatcga ggacgtcgag cccctcatgc 9361 acagcatcgc ggccggggtg gagttcatcg aggtctacgg cagcgacagc agtccttttc 9421 catctgagtt gctggatctg tgcgggcggc agaacatacc ggtccgcctc atcgactcct 9481 cgatcgtcaa ccagttgttc aagggggagc ggaaggccaa gacattcggc atcgcccgcg 9541 tccctcgccc ggccaggttc ggcgatatcg cgagccggcg tggggacgtc gtcgttctcg 9601 acggggtgaa gatcgtcggg aacatcggcg cgatagtacg cacgtcgctc gcgctcggag 9661 cgtcggggat catcctggtc gacagtgaca tcaccagcat cgcggaccgg cgtctccaaa 9721 gggccagccg aggttacgtc ttctcccttc ccgtcgttct ctccggtcgc gaggaggcca 9781 tcgccttcat tcgggacagc ggtatgcagc tgatgacgct caaggcggat ggcgacattt 9841 ccgtgaagga actcggggac aatccggatc ggctggcctt gctgttcggc agcgaaaagg 9901 gtgggccttc cgacctgttc gaggaggcgt cttccgcctc ggtttccatc cccatgatga 9961 gccagaccga gtctctcaac gtttccgttt ccctcggaat cgcgctgcac gagaggatcg 10021 acaggaatct cgcggccaac cgataagcgc ctctgttcct cggacgctcg gttgctcgac 10081 ctcgattcgt cagtgatgat caacaggacc cagaaatggc acgagcccgg gattggcctc 10141 ccgggctcgc cgcccccgaa ccggcgcagc atgccgagtg aggacgtttg ctctgtgacg 10201 agcgtacgag agaacagcgc gtcgacacca ccgcgccccg ccggaccgaa gtgccactgc 10261 ggcaccccga tcgtccacat ccccgggaag cgccccaaaa tctactgctc ggacgcctgc 10321 cgaaagcgcg caaagcgtct cattgcggac cagtcacgca gcgtagcaag cacgactaca 10381 gcgggtaatc cggacgcggg ctttagtagt aagggaaaat ctccgtccac tgcaggtcag 10441 agcgctgtaa ctggaacggg ttctacccca aacgagggtg ccgcgacagc cgggcgggag 10501 gacccgcgcg acgagcggtt cgcccggcgg gaccggcacc agacggtgtc gctgaacgag 10561 gcgttcaagg gctgcgggac ccggctcact gccgggacgg ccgagctgat gtggaagccg 10621 ggcgaggcga cgtggggcaa cacgtgccgc tgcaacaacg tccacacctg cccgtggtgc 10681 atgagccgca tcctggccgt acggggctcg aacgtgcagc tcgccgccga cggtctcgcg 10741 gacgccgggt acgggctgca ccttggcacg aacacgttgc gccacttcga gcgcatggcg 10801 ttcggcaccg tccgcaaggg aatgcggcac ggcctggtcg cggtcttgca cgacggctgg 10861 aagggcgcgt acggctcgtc cggccgccgc tggcgcacga tgcgcgacga cttcgggatc 10921 atcggctacg agcgggcctt cgaggacacc ttcggctggg gctccggctg gcacctgcac 10981 tggcacacgc tctgggtcac ccgcgaggtg ctcggcccgg acgcccaggc cgcgttccgc 11041 gacgcactcg ccggagcctg ggccgccggg gtcgaatccg cgggcggcta caccgtcagc 11101 gagacgtgcg accggcccgg ttgctcctgt gagggcaagg gccacggcac cgacgtgcgc 11161 ccactcaacg gcgcggacgc cgctgacggc gatgcgggca agcaggcccg ctacctctac 11221 aaggacggcg acaagaccaa gggcggcgtc gccaagatcg gcctggagct ggccggacag 11281 aacttcaagg ccggtcgcgg cgacgaccgc atgggaccgc tcgacctggg cgacgcggcg 11341 gccgccgagc tgcagcggct tcgtcggccc ggcccgttcg tcgagaagta ccgcgagcgc 11401 gagttcggcg tcttccaggt ccgcaagcac taccggtcgc agaacctgaa ccggctgatc 11461 aaggaactcg gcatccagca ggacgtccgc accgaggagg agatcaccga cgacaccgag 11521 ggcctggtcg ccatcgcggt catcccggcc tacatctggt accgatacat cgcccgcgtc 11581 gccggacgtc gcctcgacct catcaaggtc gccgagacgt acggcctacc gggcgtccgg 11641 cggctcgtcg agtcctgggg cctggtgtgg ggcaaggacg tcctggaccc gcccgccccg 11701 gaagcgccgg ccgcgccggg cgaccttgac gctgaccaga tgaggttcga ggtcatgtcc 11761 gaggaggaag ctgctttccg cgaggcgcgc cggaaggcca acgaggcccg cacggaggag 11821 ctggccgctt cgctcgaccg agtgcgtcag ccgaagaagg aggcgatccg cccgacgatc 11881 agccttcgga agcgcctgaa gccgaagccg gtcaccgtcg acgtcaagac tcccccgccc 11941 ggcgccgcgt cgcccgtgtg ccgccgctgc aagggcaagc tcgccccggt actccagccc 12001 tggggccagc accccggaga ctgcctccgc gtcgacacag cggtcgcgta gggccgctga 12061 gggaccgcca tgagaccgcc accccggaca tccgggtggc ggtctcatgg cggtcccggc 12121 gcgaaagccg agggtctgcc tgccgtgagg tggccggcgg acctgggtta ctgctggaat 12181 cccgcgccca gtccgagcgt ccgccggcgc ttttcacctg gctccgggtg ctcgtcgatg 12241 agcacca //