LOCUS Exported 11706 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Genome editing for gene xylR (cbei-2385) in clostridium beijerinckii NCIMB 8052. ACCESSION . VERSION . KEYWORDS pNICKclos2.0 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11706) AUTHORS Li Q, Chen J, Minton NP, Zhang Y, Wen Z, Liu J, Yang H, Zeng Z, Ren X, Yang J, Gu Y, Jiang W, Jiang Y, Yang S TITLE CRISPR-based genome editing and expression control systems in Clostridium acetobutylicum and Clostridium beijerinckii. JOURNAL Biotechnol J. 2016 May 23. doi: 10.1002/biot.201600053. PUBMED 27213844 REFERENCE 2 (bases 1 to 11706) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11706 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1488..1522 /label=J23119(SpeI) promoter /note="bacterial promoter (Registry of Standard Biological Parts BBa_J23119) modified to end with an SpeI site" misc_RNA 1543..1618 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" CDS 3804..7910 /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" primer_bind complement(7927..7944) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(7927..7943) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7936..7958) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(8152..8171) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 8271..8293 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(8331..8349) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 8417..8521 /gene="bla" /label=AmpR promoter CDS 8522..9382 /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(8740..8759) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 9553..10141 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 10042..10061 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 10295..10312 /label=L4440 /note="L4440 vector, forward primer" protein_bind 10429..10450 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 10465..10495 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 10503..10519 /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 10508..10530 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 10527..10543 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 10527..10543 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 cgataattta cagaaaagaa aattatagaa tttagtatga ttaattatac tcatttatga 61 atgtttaatt gaatacaaaa aaaaantact tgttatgtat tcaattacgg gttaaaatat 121 agacaagttg aaaaatttaa taaaaaaata agtcctcagc tcttatatat taagctacca 181 acttagtata taagccaaaa cttaaatgtg ctaccaacac atcaagccgt tagagaactc 241 tatctatagc aatatttcaa atgtaccgac atacaagaga aacattaact atatatattc 301 aatttatgag attatcttaa cagatataaa tgtaaattgc aataagtaag atttagaagt 361 ttatagcctt tgtgtattgg aagcagtacg caaaggcttt tttatttgat aaaaattaga 421 agtatattta ttttttcata attaatttat gaaaatgaaa gggggtgagc aaagtgacag 481 aggaaagcag tatcttatca aataacaagg tattagcaat atcattattg actttagcag 541 taaacattat gacttttata gtgcttgtag ctaagtagta cgaaaggggg agctttaaaa 601 agctccttgg aatacataga attcataaat taatttatga aaagaagggc gtatatgaaa 661 acttgtaaaa attgcaaaga gtttattaaa gatactgaaa tatgcaaaat acattcgttg 721 atgattcatg ataaaacagt agcaacctat tgcagtaaat acaatgagtc aagatgttta 781 cataaaggga aagtccaatg tattaattgt tcaaagatga accgatatgg atggtgtgcc 841 ataaaaatga gatgttttac agaggaagaa cagaaaaaag aacgtacatg cattaaatat 901 tatgcaagga gctttaaaaa agctcatgta aagaagagta aaaagaaaaa ataatttatt 961 tattaattta atattgagag tgccgacaca gtatgcacta aaaaatatat ctgtggtgta 1021 gtgagccgat acaaaaggat agtcactcgc attttcataa tacatcttat gttatgatta 1081 tgtgtcggtg ggacttcacg acgaaaaccc acaataaaaa aagagttcgg ggtagggtta 1141 agcatagttg aggcaactaa acaatcaagc taggatatgc agtagcagac cgtaaggtcg 1201 ttgtttaggt gtgttgtaat acatacgcta ttaagatgta aaaatacgga taccaatgaa 1261 gggaaaagta taatttttgg atgtagtttg tttgttcatc tatgggcaaa ctacgtccaa 1321 agccgtttcc aaatctgcta aaaagtatat cctttctaaa atcaaagtca agtatgaaat 1381 cataaataaa gtttaatttt gaagttatta tgatattatg tttttctatt aaaataaatt 1441 aagtatatag aatagtttaa taatagtata tacttaatgt gctgcagttg acagctagct 1501 cagtcctagg tataatacta gtcgagttag acataatagt gagttttaga gctagaaata 1561 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt 1621 ttttgcggcc gcaaaacctg gttctaatgg aattgtttac ttaccatatc ttatgggaga 1681 aagaacacct catatagacc ctaatgtaaa aggtgcattt ttaggagtat cactaataaa 1741 taaccatgat gattttgtac gtagtatatt agaaggtgtt agctttagtt tgaaaaactg 1801 ccttgattta atcgaagaca tgaatgtaaa cattgatgaa ataagagtaa gtggcggtgg 1861 agcagaaagt gctgtatgga gacaaatatt agctgatatt ttccaatatt cattaactac 1921 agttaaagca tctgaaggcg gtgcgcttgg agttgcaata ttagcagggg ttggagcagg 1981 aatctatgat tctgtagaag atgcttgtga tagaatcgta aaaggaaaca agaaagtttc 2041 tccaaatgaa gaattaaaag aagtatactc aaaagtatat gaaacatata attcagctta 2101 tccaagaata aaagatattt aaatatatta aagatagaaa tcaatcatct tagttaagta 2161 attgtataat taattaaggg tgtgtagtag tatcgaattt tagcaaaaaa atcatatagt 2221 tttttaaata ctttaacaaa aataataata gtgatattat ttaagaaaaa gctactatta 2281 ctgataatag tcgagaggag tattttaaag tttgatagaa gtaaatcaca gtaaaataaa 2341 agaaactaac agaaaaaaga taatagcatt attattaaag aaaaatgaga tcaccaaatt 2401 agatatatct agaatattag atataagtat aacaacagtt tctacaaata ttacagaatt 2461 gaaaagcgaa ggtatnnnnn nnnatgtaag atcgttggaa tctacaggag gaagaaaagc 2521 aattgctatt aagttgaatg aaaactgtag atattcaatt ggagtcgcac taacaccgaa 2581 tcatataaag atatcattag taaatataaa gaaaaaagtt attgaaagta tgctgcagtg 2641 gaatagagaa cataataaat ttacttaatg aaaatataga tttgttaatg aagaaatata 2701 atttatgctc tgcaaattta ttaggaatag ggatttcatt acctggaacc gtagatttta 2761 aagaaggcat aataaagtat tcatatttgt taggcgccaa agattttaat ttaaaggaaa 2821 agttcgaata cttagacatt ccagtatatg tagataatga agcaaattta tctgcttact 2881 acgagttttt aaataaaaga gatatactaa gaaatctatt atatgtatca ataacagaag 2941 gtttaggcct tggtataata ataaatggaa aaatttacag aggtgacaat aactcatcag 3001 gagaattagg tcatactaag atagcaattg atggaaaaaa atgtaagtgt ggagctagag 3061 gctgtttaga agcatacaca tctatgaatt ctctgattga tagttataat gaagctaata 3121 gttcaaatat tagtgatatt gatgaatttg aagaaagata taatcaaaat gacaaagatg 3181 ctcatgatgt attaaatgat tatttaaaaa cattgtcttt aggtctatcg aatttggtta 3241 tgatatttga tcctagcagc atagtaatag gtggagatat aaataattta cttaatgata 3301 agatagatat gctgaaaaaa gaggtttata aagataattt atttacaaat gaaaataatt 3361 gcagtataag tattgcaagt tttaaagaat cttatctact aggagctgct atgatgccta 3421 tagaagaatt cttagaaatt aaataatgga gtggtgaaag ctgctctatt aatttaaaaa 3481 cttattaaat aaatttttaa aatatatttt taaagttatg taagcatagc tatattaagg 3541 ggggtcttaa tctttgattg gaactccaga ttaagacaag ccttttatat aaaactaaat 3601 atctatttat ttcatattag atttatgttc ttgattttct cgagttttta acaaaatata 3661 ttgataaaaa taataatagt gggtataatt aagttgttag agaaaacgta taaattaggg 3721 ataaactatg gaacttatga aatagattga aatggtttat ctgttacccc gtatcaaaat 3781 ttaggaggtt agttagaccc gggatggata agaaatactc aataggctta gctatcggca 3841 caaatagcgt cggatgggcg gtgatcactg atgaatataa ggttccgtct aaaaagttca 3901 aggttctggg aaatacagac cgccacagta tcaaaaaaaa tcttataggg gctcttttat 3961 ttgacagtgg agagacagcg gaagcgactc gtctcaaacg gacagctcgt agaaggtata 4021 cacgtcggaa gaatcgtatt tgttatctac aggagatttt ttcaaatgag atggcgaaag 4081 tagatgatag tttctttcat cgacttgaag agtctttttt ggtggaagaa gacaagaagc 4141 atgaacgtca tcctattttt ggaaatatag tagatgaagt tgcttatcat gagaaatatc 4201 caactatcta tcatctgcga aaaaaattgg tagattctac tgataaagcg gatttgcgct 4261 taatctattt ggccttagcg catatgatta agtttcgtgg tcattttttg attgagggag 4321 atttaaatcc tgataatagt gatgtggaca aactatttat ccagttggta caaacctaca 4381 atcaattatt tgaagaaaac cctattaacg caagtggagt agatgctaaa gcgattcttt 4441 ctgcacgatt gagtaaatca agacgattag aaaatctcat tgctcagctc cccggtgaga 4501 agaaaaatgg cttatttggg aatctcattg ctttgtcatt gggtttgacc cctaatttta 4561 aatcaaattt tgatttggca gaagatgcta aattacagct ttcaaaagat acttacgatg 4621 atgatttaga taatttattg gcgcaaattg gagatcaata tgctgatttg tttttggcag 4681 ctaagaattt atcagatgct attttacttt cagatatcct aagagtaaat actgaaataa 4741 ctaaggctcc cctatcagct tcaatgatta aacgctacga tgaacatcat caagacttga 4801 ctcttttaaa agctttagtt cgacaacaac ttccagaaaa gtataaagaa atcttttttg 4861 atcaatcaaa aaacggatat gcaggttata ttgatggggg agctagccaa gaagaatttt 4921 ataaatttat caaaccaatt ttagaaaaaa tggatggtac tgaggaatta ttggtgaaac 4981 taaatcgtga agatttgctg cgcaagcaac ggacctttga caacggctct attccccatc 5041 aaattcactt gggtgagctg catgctattt tgagaagaca agaagacttt tatccatttt 5101 taaaagacaa tcgtgagaag attgaaaaaa tcttgacttt tcgaattcct tattatgttg 5161 gtccattggc gcgtggcaat agtcgttttg catggatgac tcggaagtct gaagaaacaa 5221 ttaccccatg gaattttgaa gaagttgtcg ataaaggtgc ttcagctcaa tcatttattg 5281 aacgcatgac aaactttgat aaaaatcttc caaatgaaaa agtactacca aaacatagtt 5341 tgctttatga gtattttacg gtttataacg aattgacaaa ggtcaaatat gttactgaag 5401 gaatgcgaaa accagcattt ctttcaggtg aacagaagaa agccattgtt gatttactct 5461 tcaaaacaaa tcgaaaagta accgttaagc aattaaaaga agattatttc aaaaaaatag 5521 aatgttttga tagtgttgaa atttcaggag ttgaagatag atttaatgct tcattaggta 5581 cctaccatga tttgctaaaa attattaaag ataaagattt tttggataat gaagaaaatg 5641 aagatatctt agaggatatt gttttaacat tgaccttatt tgaagatagg gagatgattg 5701 aggaaagact taaaacatat gctcacctct ttgatgataa ggtgatgaaa cagcttaaac 5761 gtcgccgtta tactggttgg ggacgtttgt ctcgaaaatt gattaatggt attagggata 5821 agcaatctgg caaaacaata ttagattttt tgaaatcaga tggttttgcc aatcgcaatt 5881 ttatgcagct gatccatgat gatagtttga catttaaaga agacattcaa aaagcacaag 5941 tgtctggaca aggcgatagt ttacatgaac atattgcaaa tttagctggt agccctgcta 6001 ttaaaaaagg tattttacag actgtaaaag ttgttgatga attggtcaaa gtaatggggc 6061 ggcataagcc agaaaatatc gttattgaaa tggcacgtga aaatcagaca actcaaaagg 6121 gccagaaaaa ttcgcgagag cgtatgaaac gaatcgaaga aggtatcaaa gaattaggaa 6181 gtcagattct taaagagcat cctgttgaaa atactcaatt gcaaaatgaa aagctctatc 6241 tctattatct ccaaaatgga agagacatgt atgtggacca agaattagat attaatcgtt 6301 taagtgatta tgatgtcgat cacattgttc cacaaagttt ccttaaagac gattcaatag 6361 acaataaggt cttaacgcgt tctgataaaa atcgtggtaa atcggataac gttccaagtg 6421 aagaagtagt caaaaagatg aaaaactatt ggagacaact tctaaacgcc aagttaatca 6481 ctcaacgtaa gtttgataat ttaacgaaag ctgaacgtgg aggtttgagt gaacttgata 6541 aagctggttt tatcaaacgc caattggttg aaactcgcca aatcactaag catgtggcac 6601 aaattttgga tagtcgcatg aatactaaat acgatgaaaa tgataaactt attcgagagg 6661 ttaaagtgat taccttaaaa tctaaattag tttctgactt ccgaaaagat ttccaattct 6721 ataaagtacg tgagattaac aattaccatc atgcccatga tgcgtatcta aatgccgtcg 6781 ttggaactgc tttgattaag aaatatccaa aacttgaatc ggagtttgtc tatggtgatt 6841 ataaagttta tgatgttcgt aaaatgattg ctaagtctga gcaagaaata ggcaaagcaa 6901 ccgcaaaata tttcttttac tctaatatca tgaacttctt caaaacagaa attacacttg 6961 caaatggaga gattcgcaaa cgccctctaa tcgaaactaa tggggaaact ggagaaattg 7021 tctgggataa agggcgagat tttgccacag tgcgcaaagt attgtccatg ccccaagtca 7081 atattgtcaa gaaaacagaa gtacagacag gcggattctc caaggagtca attttaccaa 7141 aaagaaattc ggacaagctt attgctcgta aaaaagactg ggatccaaaa aaatatggtg 7201 gttttgatag tccaacggta gcttattcag tcctagtggt tgctaaggtg gaaaaaggga 7261 aatcgaagaa gttaaaatcc gttaaagagt tactagggat cacaattatg gaaagaagtt 7321 cctttgaaaa aaatccgatt gactttttag aagctaaagg atataaggaa gttaaaaaag 7381 acttaatcat taaactacct aaatatagtc tttttgagtt agaaaacggt cgtaaacgga 7441 tgctggctag tgccggagaa ttacaaaaag gaaatgagct ggctctgcca agcaaatatg 7501 tgaatttttt atatttagct agtcattatg aaaagttgaa gggtagtcca gaagataacg 7561 aacaaaaaca attgtttgtg gagcagcata agcattattt agatgagatt attgagcaaa 7621 tcagtgaatt ttctaagcgt gttattttag cagatgccaa tttagataaa gttcttagtg 7681 catataacaa acatagagac aaaccaatac gtgaacaagc agaaaatatt attcatttat 7741 ttacgttgac gaatcttgga gctcccgctg cttttaaata ttttgataca acaattgatc 7801 gtaaacgata tacgtctaca aaagaagttt tagatgccac tcttatccat caatccatca 7861 ctggtcttta tgaaacacgc attgatttga gtcagctagg aggtgactga gcggccgcgg 7921 gaattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact 7981 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac 8041 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt 8101 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg 8161 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg 8221 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg 8281 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat 8341 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac 8401 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat 8461 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag 8521 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc 8581 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc 8641 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc 8701 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc 8761 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt 8821 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt 8881 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat 8941 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct 9001 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat 9061 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc 9121 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg 9181 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc 9241 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta 9301 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc 9361 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga 9421 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat 9481 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat 9541 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa 9601 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa 9661 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt 9721 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt 9781 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata 9841 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt 9901 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac 9961 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga 10021 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg 10081 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa 10141 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat 10201 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc 10261 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga 10321 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg 10381 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta 10441 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg 10501 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct 10561 ttggctaaca cacacgccat tccaaccaat agttttctcg gcataaagcc atgctctgac 10621 gcttaaatgc actaatgcct taaaaaaaca ttaaagtcta acacactaga cttatttact 10681 tcgtaattaa gtcgttaaac cgtgtgctct acgaccaaaa gtataaaacc tttaagaact 10741 ttcttttttc ttgtaaaaaa agaaactaga taaatctctc atatctttta ttcaataatc 10801 gcatcagatt gcagtataaa tttaacgatc actcatcatg ttcatattta tcagagctcg 10861 tgctataatt atactaattt tataaggagg aaaaaataaa gagggttata atgaacgaga 10921 aaaatataaa acacagtcaa aactttatta cttcaaaaca taatatagat aaaataatga 10981 caaatataag attaaatgaa catgataata tctttgaaat cggctcagga aaagggcatt 11041 ttacccttga attagtacag aggtgtaatt tcgtaactgc cattgaaata gaccataaat 11101 tatgcaaaac tacagaaaat aaacttgttg atcacgataa tttccaagtt ttaaacaagg 11161 atatattgca gtttaaattt cctaaaaacc aatcctataa aatatttggt aatatacctt 11221 ataacataag tacggatata atacgcaaaa ttgtttttga tagtatagct gatgagattt 11281 atttaatcgt ggaatacggg tttgctaaaa gattattaaa tacaaaacgc tcattggcat 11341 tatttttaat ggcagaagtt gatatttcta tattaagtat ggttccaaga gaatattttc 11401 atcctaaacc taaagtgaat agctcactta tcagattaaa tagaaaaaaa tcaagaatat 11461 cacacaaaga taaacagaag tataattatt tcgttatgaa atgggttaac aaagaataca 11521 agaaaatatt tacaaaaaat caatttaaca attccttaaa acatgcagga attgacgatt 11581 taaacaatat tagctttgaa caattcttat ctcttttcaa tagctataaa ttatttaata 11641 agtaagttaa gggatgcata aactgcatcc cttaacttgt ttttcgtgta cctatttttt 11701 gtgaat //