LOCUS Exported 9794 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION To express the recombinant APOBEC3A in vitro. ACCESSION . VERSION . KEYWORDS pET42b-A3A-PBE-Delta-UGI SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9794) AUTHORS Zong Y, Song Q, Li C, Jin S, Zhang D, Wang Y, Qiu JL, Gao C TITLE Efficient C-to-T base editing in plants using a fusion of nCas9 and human APOBEC3A. JOURNAL Nat Biotechnol. 2018 Oct 1. pii: nbt.4261. doi: 10.1038/nbt.4261. PUBMED 30272679 REFERENCE 2 (bases 1 to 9794) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9794 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..20 /label=T7 /note="T7 promoter, forward primer" promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 20..44 /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)." RBS 59..81 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 100..117 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 766..4866 /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 4867..4914 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" primer_bind complement(4962..4980) /label=T7 Term /note="T7 terminator, reverse primer" terminator 4976..5023 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 5060..5515 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5147..5166) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5357..5378 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(5607..6422) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 6330..6349 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 6414..6433 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 6544..7132 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7033..7052 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7286..7303 /label=L4440 /note="L4440 vector, forward primer" misc_feature 7318..7457 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(7383..7402) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7543..7565 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(7559..7750) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(8322..9404) /codon_start=1 /gene="lacI" /product="lac repressor" /label=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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind 9366..9385 /label=LacI-R /note="LacI, reverse primer" promoter complement(9405..9482) /gene="lacI" /label=lacI promoter primer_bind 9688..9707 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt 61 tgtttaactt taagaaggag atatacaatg ggcagcagtc atcatcatca ccatcacgag 121 gccagcccgg ctagcggccc aaggcatctc atggacccgc acatcttcac cagcaacttc 181 aacaacggca tcggcaggca caagacctac ttgtgctacg aggtggagag gctcgacaac 241 ggaacctccg tgaagatgga ccaacacagg gggttcctcc acaaccaagc caagaacctc 301 ctctgcggct tctacggcag gcacgccgag ttgaggttcc tcgacttggt gccatccctc 361 caactcgatc cagcccaaat ctaccgcgtg acctggttca tctcctggtc cccatgcttc 421 tcctggggtt gcgccggcga ggttcgggct ttcctccaag aaaacaccca cgtccgcctc 481 cgcattttcg ccgccaggat ctatgattac gaccctctct acaaggaggc cctccagatg 541 ctgcgggacg ccggtgctca ggtgagtatc atgacctacg acgagttcaa gcactgctgg 601 gacaccttcg ttgaccacca gggctgccca ttccaaccat gggacggtct ggatgaacac 661 agccaagcct tgtccggcag gctccgggcc atcctccaaa accaggggaa ctccgggagc 721 gagacgccag gcacctccga gtcggccacc ccagaatctc ttaaggacaa gaagtactcg 781 atcggcctcg ccatcgggac gaactcagtt ggctgggccg tgatcaccga cgagtacaag 841 gtgccctcta agaagttcaa ggtcctgggg aacaccgacc gccattccat caagaagaac 901 ctcatcggcg ctctcctgtt cgacagcggg gagaccgctg aggctacgag gctcaagaga 961 accgctaggc gccggtacac gagaaggaag aacaggatct gctacctcca agagattttc 1021 tccaacgaga tggccaaggt tgacgattca ttcttccacc gcctggagga gtctttcctc 1081 gtggaggagg ataagaagca cgagcggcat cccatcttcg gcaacatcgt ggacgaggtt 1141 gcctaccacg agaagtaccc tacgatctac catctgcgga agaagctcgt ggactccacc 1201 gataaggcgg acctcagact gatctacctc gctctggccc acatgatcaa gttccgcggc 1261 catttcctga tcgaggggga tctcaaccca gacaacagcg atgttgacaa gctgttcatc 1321 caactcgtgc agacctacaa ccaactcttc gaggagaacc cgatcaacgc ctctggcgtg 1381 gacgcgaagg ctatcctgtc cgcgaggctc tcgaagtcca ggaggctgga gaacctgatc 1441 gctcagctcc caggcgagaa gaagaacggc ctgttcggga acctcatcgc tctcagcctg 1501 gggctcaccc cgaacttcaa gtcgaacttc gatctcgctg aggacgccaa gctgcaactc 1561 tccaaggaca cctacgacga tgacctcgat aacctcctgg cccagatcgg cgatcaatac 1621 gcggacctgt tcctcgctgc caagaacctg tcggacgcca tcctcctgtc agatatcctc 1681 cgcgtgaaca ccgagatcac gaaggctcca ctctctgcct ccatgatcaa gcgctacgac 1741 gagcaccatc aggatctgac cctcctgaag gcgctggtcc gccaacagct cccggagaag 1801 tacaaggaga ttttcttcga tcagtcgaag aacggctacg ctgggtacat cgacggcggg 1861 gcctcacaag aggagttcta caagttcatc aagccaatcc tggagaagat ggacggcacg 1921 gaggagctcc tggtgaagct caacagggag gacctcctgc ggaagcagag aaccttcgat 1981 aacggcagca tcccccacca aatccatctc ggggagctgc acgccatcct gagaaggcaa 2041 gaggacttct accctttcct caaggataac cgggagaaga tcgagaagat cctgaccttc 2101 agaatcccat actacgtcgg ccctctcgcg cgggggaact caagattcgc ttggatgacc 2161 cgcaagtctg aggagaccat cacgccgtgg aacttcgagg aggtggtgga caagggcgct 2221 agcgctcagt cgttcatcga gaggatgacc aacttcgaca agaacctgcc caacgagaag 2281 gtgctcccta agcactcgct cctgtacgag tacttcaccg tctacaacga gctcacgaag 2341 gtgaagtacg tcaccgaggg catgcgcaag ccagcgttcc tgtccgggga gcagaagaag 2401 gctatcgtgg acctcctgtt caagaccaac cggaaggtca cggttaagca actcaaggag 2461 gactacttca agaagatcga gtgcttcgat tcggtcgaga tcagcggcgt tgaggaccgc 2521 ttcaacgcca gcctcgggac ctaccacgat ctcctgaaga tcatcaagga taaggacttc 2581 ctggacaacg aggagaacga ggatatcctg gaggacatcg tgctgaccct cacgctgttc 2641 gaggacaggg agatgatcga ggagcgcctg aagacgtacg cccatctctt cgatgacaag 2701 gtcatgaagc aactcaagcg ccggagatac accggctggg ggaggctgtc ccgcaagctc 2761 atcaacggca tccgggacaa gcagtccggg aagaccatcc tcgacttcct caagagcgat 2821 ggcttcgcca acaggaactt catgcaactg atccacgatg acagcctcac cttcaaggag 2881 gatatccaaa aggctcaagt gagcggccag ggggactcgc tgcacgagca tatcgcgaac 2941 ctcgctggct cccccgcgat caagaagggc atcctccaga ccgtgaaggt tgtggacgag 3001 ctcgtgaagg tcatgggccg gcacaagcct gagaacatcg tcatcgagat ggccagagag 3061 aaccaaacca cgcagaaggg gcaaaagaac tctagggagc gcatgaagcg catcgaggag 3121 ggcatcaagg agctggggtc ccaaatcctc aaggagcacc cagtggagaa cacccaactg 3181 cagaacgaga agctctacct gtactacctc cagaacggca gggatatgta cgtggaccaa 3241 gagctggata tcaaccgcct cagcgattac gacgtcgatc atatcgttcc ccagtctttc 3301 ctgaaggatg actccatcga caacaaggtc ctcaccaggt cggacaagaa ccgcggcaag 3361 tcagataacg ttccatctga ggaggtcgtt aagaagatga agaactactg gaggcagctc 3421 ctgaacgcca agctgatcac gcaaaggaag ttcgacaacc tcaccaaggc tgagagaggc 3481 gggctctcag agctggacaa ggccggcttc atcaagcggc agctggtcga gaccagacaa 3541 atcacgaagc acgttgcgca aatcctcgac tctcggatga acacgaagta cgatgagaac 3601 gacaagctga tcagggaggt taaggtgatc accctgaagt ctaagctcgt ctccgacttc 3661 aggaaggatt tccagttcta caaggttcgc gagatcaaca actaccacca tgcccatgac 3721 gcttacctca acgctgtggt cggcaccgct ctgatcaaga agtacccaaa gctggagtcc 3781 gagttcgtgt acggggacta caaggtttac gatgtgcgca agatgatcgc caagtcggag 3841 caagagatcg gcaaggctac cgccaagtac ttcttctact caaacatcat gaacttcttc 3901 aagaccgaga tcacgctggc caacggcgag atccggaaga gaccgctcat cgagaccaac 3961 ggcgagacgg gggagatcgt gtgggacaag ggcagggatt tcgcgaccgt ccgcaaggtt 4021 ctctccatgc cccaggtgaa catcgtcaag aagaccgagg tccaaacggg cgggttctca 4081 aaggagtcta tcctgcctaa gcggaacagc gacaagctca tcgccagaaa gaaggactgg 4141 gacccaaaga agtacggcgg gttcgacagc cctaccgtgg cctactcggt cctggttgtg 4201 gcgaaggttg agaagggcaa gtccaagaag ctcaagagcg tgaaggagct cctggggatc 4261 accatcatgg agaggtccag cttcgagaag aacccaatcg acttcctgga ggccaagggc 4321 tacaaggagg tgaagaagga cctgatcatc aagctcccga agtactctct cttcgagctg 4381 gagaacggca ggaagagaat gctggcttcc gctggcgagc tccagaaggg gaacgagctc 4441 gcgctgccaa gcaagtacgt gaacttcctc tacctggctt cccactacga gaagctcaag 4501 ggcagcccgg aggacaacga gcaaaagcag ctgttcgtcg agcagcacaa gcattacctc 4561 gacgagatca tcgagcaaat ctccgagttc agcaagcgcg tgatcctcgc cgacgcgaac 4621 ctggataagg tcctctccgc ctacaacaag caccgggaca agcccatcag agagcaagcg 4681 gagaacatca tccatctctt caccctgacg aacctcggcg ctcctgctgc tttcaagtac 4741 ttcgacacca cgatcgatcg gaagagatac acctccacga aggaggtcct ggacgcgacc 4801 ctcatccacc agtcgatcac cggcctgtac gagacgagga tcgacctctc acaactcggc 4861 ggggataaga gacccgcagc aaccaagaag gcagggcaag caaagaagaa gaagtaggct 4921 gctaaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc 4981 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat 5041 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg 5101 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc 5161 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc 5221 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt 5281 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag 5341 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg 5401 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag 5461 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg 5521 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa 5581 atatgtatcc gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc 5641 aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa 5701 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt 5761 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca 5821 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt 5881 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca 5941 accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta 6001 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca 6061 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg 6121 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga 6181 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca 6241 acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga 6301 tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca 6361 gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc 6421 ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca tgaccaaaat 6481 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc 6541 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct 6601 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg 6661 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca 6721 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc 6781 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga 6841 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac 6901 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga 6961 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 7021 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg 7081 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag 7141 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc 7201 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc 7261 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct 7321 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc 7381 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg 7441 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt 7501 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc 7561 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt 7621 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct 7681 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct 7741 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga 7801 tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg 7861 aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc 7921 agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc 7981 atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac 8041 tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc 8101 agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc 8161 aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcta gtcatgcccc 8221 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg 8281 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt 8341 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt 8401 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg 8461 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc 8521 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta 8581 tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc ggtaatggcg 8641 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc 8701 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt 8761 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga 8821 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg 8881 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg 8941 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca 9001 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg 9061 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc 9121 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc 9181 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt 9241 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc 9301 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag 9361 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat 9421 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg 9481 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag 9541 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa 9601 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc 9661 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc 9721 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatcgatct 9781 cgatcccgcg aaat //