LOCUS Exported 11506 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION parental, all-in-one CRISPRi vector for B. burgdorferi, parental for gRNA cloning. ACCESSION . VERSION . KEYWORDS pBbdCas9S_P0526-sgRNA500 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11506) AUTHORS Takacs CN, Scott M, Chang Y, Kloos ZA, Irnov I, Rosa PA, Liu J, Jacobs-Wagner C TITLE A CRISPR interference platform for selective downregulation of gene expression in Borrelia burgdorferi. JOURNAL Appl Environ Microbiol. 2020 Nov 30. pii: AEM.02519-20. doi: 10.1128/AEM.02519-20. PUBMED 33257311 REFERENCE 2 (bases 1 to 11506) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11506 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1434..1478 /label=T5 promoter /note="bacteriophage T5 promoter for E. coli RNA polymerase, with embedded lac operator" protein_bind 1454..1470 /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)." protein_bind 1486..1502 /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 1521..1532 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 1539..5645 /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TILERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 5815..6606 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 6729..7317 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7218..7237 /label=pBR322ori-F /note="pBR322 origin, forward primer" misc_RNA 11422..11497 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" ORIGIN 1 cgcgccaaat aatgcccgtg tccctgttga atagggacac ggttttttaa caattttttt 61 attcctaggc cttagtggcc ggtacccggg gatccttatt actggccgct ttctagcctg 121 gatacttgtc tggctagttg cattagagag tcagctagag ccctagggct agctgtttgt 181 gtattcggag ccagtgtagt ttttcttttt actaacgata cgggtaataa ttgattgcct 241 tttacagcct gaccttgaga caattgcaaa agtctgtcca cagaggtttg acctaataat 301 ctaaaatctt gtttgatagt agtaagtgga ggtatgtagc aactagaatc ttctgtatca 361 tcataaccca caactgaaat atcagcccca actctaagtc cactttcagt aatagctcgc 421 atagctccta aagccatttg atcattagca actaacatag cagtagggac aatgccttca 481 tttaacattt gcattgtttg ctgaaatcct gacatagcag accaatctcc ttctctttct 541 gctattggtt gtatttgatt tcttgttaaa tacttgtgcc atcctgctaa tctcaatctg 601 gcacttactg aacttaaggg acctgcaagt agtgcaattt gttgatggcc taatgcaaca 661 aggtgttcaa ctcctaacct tgttccatct tcgtgagaaa aaattataga gtttattggg 721 gtttggtccg acacatccaa aaacaatgct ggaacattag tacatgctgc ttcaactgca 781 atcgcatctt ggtcatcaag aggataatta attataagtc cactaacacg ttgagctaat 841 aaattatgta ctgctgcctt acaagcctct acacctgatc tctcaaccat tgatacaaca 901 acgcttgcac caagttgatc agctctcgat ttaattgcgg ctacaatttg actaggcgca 961 tgtaatgcaa gtgatgatgt ggcaactcca attaaaagtg attgtttccc cgcaagttgc 1021 tgagcaacgc gatttggtat ataattcaac tctgccatag ctgcttcaac cttctctctt 1081 gtttttgcag atacatgact tgcttggttc acaactctcg aaaccgtttg gtagctaacc 1141 ccggcatatt cagcaacatc atacaaagta acaggtttca tggatcattc ctccatgata 1201 aaatttaaat ttctgacttt ggcaaatcct tttgcctgtt aaaaagaata gcacaaaaat 1261 taaaaaaaaa gaaacgcaaa tagcaatagc attaaaaata attattatta tttatttctt 1321 tgattaaaat caaaagcctt taatgtattt tcttgatctt tcatgcgctt aacggtaaaa 1381 tccaaggtat taagttcacc ttccactttt ctctctctat cttctctaga aaatcataaa 1441 aaatttattt gctttgtgag cggataacaa ttataataga ttcaattgtg agcggataac 1501 aatttcacac agaattcatt aaagaggaga aattacatat ggataagaaa tactcaatag 1561 gcttagctat cggcacaaat agcgtcggat gggcggtgat cactgatgaa tataaggttc 1621 cgtctaaaaa gttcaaggtt ctgggaaata cagaccgcca cagtatcaaa aaaaatctta 1681 taggggctct tttatttgac agtggagaga cagcggaagc gactcgtctc aaacggacag 1741 ctcgtagaag gtatacacgt cggaagaatc gtatttgtta tctacaggag attttttcaa 1801 atgagatggc gaaagtagat gatagtttct ttcatcgact tgaagagtct tttttggtgg 1861 aagaagacaa gaagcatgaa cgtcatccta tttttggaaa tatagtagat gaagttgctt 1921 atcatgagaa atatccaact atctatcatc tgcgaaaaaa attggtagat tctactgata 1981 aagcggattt gcgcttaatc tatttggcct tagcgcatat gattaagttt cgtggtcatt 2041 ttttgattga gggagattta aatcctgata atagtgatgt ggacaaacta tttatccagt 2101 tggtacaaac ctacaatcaa ttatttgaag aaaaccctat taacgcaagt ggagtagatg 2161 ctaaagcgat tctttctgca cgattgagta aatcaagacg attagaaaat ctcattgctc 2221 agctccccgg tgagaagaaa aatggcttat ttgggaatct cattgctttg tcattgggtt 2281 tgacccctaa ttttaaatca aattttgatt tggcagaaga tgctaaatta cagctttcaa 2341 aagatactta cgatgatgat ttagataatt tattggcgca aattggagat caatatgctg 2401 atttgttttt ggcagctaag aatttatcag atgctatttt actttcagat atcctaagag 2461 taaatactga aataactaag gctcccctat cagcttcaat gattaaacgc tacgatgaac 2521 atcatcaaga cttgactctt ttaaaagctt tagttcgaca acaacttcca gaaaagtata 2581 aagaaatctt ttttgatcaa tcaaaaaacg gatatgcagg ttatattgat gggggagcta 2641 gccaagaaga attttataaa tttatcaaac caattttaga aaaaatggat ggtactgagg 2701 aattattggt gaaactaaat cgtgaagatt tgctgcgcaa gcaacggacc tttgacaacg 2761 gctctattcc ccatcaaatt cacttgggtg agctgcatgc tattttgaga agacaagaag 2821 acttttatcc atttttaaaa gacaatcgtg agaagattga aaaaatcttg acttttcgaa 2881 ttccttatta tgttggtcca ttggcgcgtg gcaatagtcg ttttgcatgg atgactcgga 2941 agtctgaaga aacaattacc ccatggaatt ttgaagaagt tgtcgataaa ggtgcttcag 3001 ctcaatcatt tattgaacgc atgacaaact ttgataaaaa tcttccaaat gaaaaagtac 3061 taccaaaaca tagtttgctt tatgagtatt ttacggttta taacgaattg acaaaggtca 3121 aatatgttac tgaaggaatg cgaaaaccag catttctttc aggtgaacag aagaaagcca 3181 ttgttgattt actcttcaaa acaaatcgaa aagtaaccgt taagcaatta aaagaagatt 3241 atttcaaaaa aatagaatgt tttgatagtg ttgaaatttc aggagttgaa gatagattta 3301 atgcttcatt aggtacctac catgatttgc taaaaattat taaagataaa gattttttgg 3361 ataatgaaga aaatgaagat atcttagagg atattgtttt aacattgacc ttatttgaag 3421 atagggagat gattgaggaa agacttaaaa catatgctca cctctttgat gataaggtga 3481 tgaaacagct taaacgtcgc cgttatactg gttggggacg tttgtctcga aaattgatta 3541 atggtattag ggataagcaa tctggcaaaa caatattaga ttttttgaaa tcagatggtt 3601 ttgccaatcg caattttatg cagctgatcc atgatgatag tttgacattt aaagaagaca 3661 ttcaaaaagc acaagtgtct ggacaaggcg atagtttaca tgaacatatt gcaaatttag 3721 ctggtagccc tgctattaaa aaaggtattt tacagactgt aaaagttgtt gatgaattgg 3781 tcaaagtaat ggggcggcat aagccagaaa atatcgttat tgaaatggca cgtgaaaatc 3841 agacaactca aaagggccag aaaaattcgc gagagcgtat gaaacgaatc gaagaaggta 3901 tcaaagaatt aggaagtcag attcttaaag agcatcctgt tgaaaatact caattgcaaa 3961 atgaaaagct ctatctctat tatctccaaa atggaagaga catgtatgtg gaccaagaat 4021 tagatattaa tcgtttaagt gattatgatg tcgatgccat tgttccacaa agtttcctta 4081 aagacgattc aatagacaat aaggtcttaa cgcgttctga taaaaatcgt ggtaaatcgg 4141 ataacgttcc aagtgaagaa gtagtcaaaa agatgaaaaa ctattggaga caacttctaa 4201 acgccaagtt aatcactcaa cgtaagtttg ataatttaac gaaagctgaa cgtggaggtt 4261 tgagtgaact tgataaagct ggttttatca aacgccaatt ggttgaaact cgccaaatca 4321 ctaagcatgt ggcacaaatt ttggatagtc gcatgaatac taaatacgat gaaaatgata 4381 aacttattcg agaggttaaa gtgattacct taaaatctaa attagtttct gacttccgaa 4441 aagatttcca attctataaa gtacgtgaga ttaacaatta ccatcatgcc catgatgcgt 4501 atctaaatgc cgtcgttgga actgctttga ttaagaaata tccaaaactt gaatcggagt 4561 ttgtctatgg tgattataaa gtttatgatg ttcgtaaaat gattgctaag tctgagcaag 4621 aaataggcaa agcaaccgca aaatatttct tttactctaa tatcatgaac ttcttcaaaa 4681 cagaaattac acttgcaaat ggagagattc gcaaacgccc tctaatcgaa actaatgggg 4741 aaactggaga aattgtctgg gataaagggc gagattttgc cacagtgcgc aaagtattgt 4801 ccatgcccca agtcaatatt gtcaagaaaa cagaagtcca gacaggcgga ttctccaagg 4861 agtcaatttt accaaaaaga aattcggaca agcttattgc tcgtaaaaaa gactgggatc 4921 caaaaaaata tggtggtttt gatagtccaa cggtagctta ttcagtccta gtggttgcta 4981 aggtggaaaa agggaaatcg aagaagttaa aatccgttaa agagttacta gggatcacaa 5041 ttctggaaag aagttccttt gaaaaaaatc cgattgactt tttagaagct aaaggatata 5101 aggaagttaa aaaagactta atcattaaac tacctaaata tagtcttttt gagttagaaa 5161 acggtcgtaa acggatgctg gctagtgccg gagaattaca aaaaggaaat gagctggctc 5221 tgccaagcaa atatgtgaat tttttatatt tagctagtca ttatgaaaag ttgaagggta 5281 gtccagaaga taacgaacaa aaacaattgt ttgtggagca gcataagcat tatttagatg 5341 agattattga gcaaatcagt gaattttcta agcgtgttat tttagcagat gccaatttag 5401 ataaagttct tagtgcatat aacaaacata gagacaaacc aatacgtgaa caagcagaaa 5461 atattattca tttatttacg ttgacgaatc ttggagctcc cgctgctttt aaatattttg 5521 atacaacaat tgatcgtaaa cgatatacgt ctacaaaaga agttttagat gccactctta 5581 tccatcaatc catcactggt ctttatgaaa cacgcattga tttgagtcag ctaggaggtg 5641 actaataaaa gcttgatcag atctcagctt ttttttgaag tgcctggcag taagttgaaa 5701 aaatttgaaa taaacttgga ctatgttaat aaaaaaatac aaattaacga acaaaaagat 5761 ttgaattatt atataattta gttaaaagca attttaaatg agggaggttt ccatatgagg 5821 gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc 5881 catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg 5941 aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg 6001 cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt 6061 ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca 6121 gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc 6181 gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc 6241 gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta 6301 tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat 6361 gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc 6421 gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc 6481 gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc 6541 gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc 6601 aaataatgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcatgacc 6661 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 6721 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 6781 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 6841 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 6901 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 6961 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 7021 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 7081 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 7141 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 7201 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 7261 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 7321 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 7381 tttcctgcac tagatccccg gcaagttcat ccttagggtt ttcatatctc ttaaaaagtc 7441 aatttctgct tgattaattt cttgtggatt attgtttttg cggtttttca ttttttatct 7501 ccttgagtat tgaatttttt cttatatata aatatatagc aaaaactatt tttgccaact 7561 tttttacaaa aaatttttta ttaaaacact tagggctttg ctaaattctc ttttaaaaga 7621 acttagtaaa gccctatgga tttaagaact gattatactt acgtagtaaa aatactacag 7681 atattaatag tatagaccat atcaaaatca taatcaattt atatttatct aaaattgtag 7741 tccatacaaa atacaaaaaa attaggtttt tctaaattct attttaaaag aacttagtaa 7801 cacctaatct ctaaattact tacaaaattc taaaattaga tttttagttc ttcatatttc 7861 tttaaaagtt ttttaagaaa atctttttga ttttcaaaaa tttcatgcat cataaaacta 7921 gtaaatttgg attttttttt gtaaaaatca taactttctt gggttttaag ttgaaatctt 7981 aatggtttta ttggattttg cctagatttt tttaacgccg gactttcttt atcttttata 8041 aaattataag aatttttaat accattttct attaaataat cttcttttat aagcccctct 8101 tgcagtgcag ttgctaatct taaatactta tacgcttgag tttttgctac atcaaagtct 8161 tttataaaat caagaaaagt tttataacca tcaaatttat aatattgatt atcttttatt 8221 tcttttaaaa ttttaatatt ttccaccctg tggaatattt cttttttgaa attgtcattt 8281 aacttttcct ttagtgtctt gtaataagaa attttacttt cattaatcat taaattttgt 8341 tcaacataag tatctatctt ttttgttaaa actttaacaa cacgtttttt ttccataaaa 8401 attttccttt atttttgcaa cattttcctt aattgatata attgttacta ttttagattt 8461 tttaatatcc acttaagaat taatccctca ctagaatatt ttttattata gaattcatat 8521 acatgttttt ctaaatctat aaatttatca acaattgctt tattataggg tgtatctatc 8581 ttttcttttc ttaataatcg tctcatattc tttaaataac aacaaacact gccggtttta 8641 aacctaaatt ctatgtaata tatttttgaa aaagtatata cattttcaac cccgtttatt 8701 tcatacttta caaaaacatt atttattggt tttctataac catagtaaat gcctaaaaat 8761 ttgtccccgt ctttcatagg atatagataa agtttttgaa tctctgattt atctaataat 8821 ttttcaaaaa aaattataca ccgcttttgt ttgttactaa ctactctatg caatgccttt 8881 aatatttttg tgtgatatat atttttatca cctttttttt ctttcagcag gaagatactt 8941 tcttttggaa atttaatttt ttgttcattt attcttgcag ccttaagtcc gattttcata 9001 tttaatcctt atatagcctt tctatttaat tcttttaaac tcaataaact tttttggttt 9061 ttcattagtt ctaataattc gcaataatat gtgcaaaaca ctttattgta ttctattttc 9121 ttttggttat tcaaatattc ttttataatt ggctttagaa gttcaatatt tacttctttt 9181 tttagttgtt caataaggat attgaaaatg tttgttttta aatcttgata attttgttga 9241 gaattttctt tttttctttc aattgacttc tctaatttac gctttatatt gtttaaatcg 9301 ctatatttat gactttcaat aataaaatga ggcttaaatt tgtaattttc gtatattttt 9361 tggaaacttg tttctaattg tttagaatta tattcatttt tttcaaattc tttttgagtg 9421 ttgcatagaa tttcctttaa tttttcttgt ttttctttca tacaagattt agtaaaatgt 9481 atatttttat tttttagtaa tttaatttct attattttta tgattttaag tattttgatt 9541 atattatctt tatcaatatt taaattgaat aaaattgtaa gaaaaattga aagaattttt 9601 ttacataaaa agttacattt attgaagtaa tttcttactt gatacttttc tatttcatta 9661 atctttcttt cttcttttat attattttta ttatttaaac actcctttaa atctacactc 9721 ccatttttag taaatctatc tttaaggccc cttgaaactc tattttgaaa tcttaaatct 9781 tttttttctt gaaagtattt atagattttt tggtaacatt ctttttttga aaaattaagt 9841 tgatagtata tttcggtccc caaatttacc cctaagtgtt tgtgataatt agttgtgact 9901 ttaaattctt tttctaattt gtaaaggtaa ttttgcagtg tttttagttt aacgggtgtt 9961 tgactatttc ttttaaggtt ttcattaaag taatagagta tgtttttttg tgtgtatttc 10021 ttatatgttt tatttatgta cgctagtgtt gagataagga ctactaattt atactgtaat 10081 ttattgtagc aagttgtggt tttactggtt ttgtttatat tagataagtt ttccatttat 10141 agctccttga ataccggatt ttttatctat aaagtatata tatcacaatt ttgatttaaa 10201 agtaaattct ttttttaaaa atacaaaaat tttttataaa ttatatagac catgatattg 10261 tattaatact aattgatttt aatttaaggt tgaactaaaa taaacatagt tttgtagaaa 10321 ataatttttc attattttgc tacttgatat tggatcgtaa aaatattagg gctttacaaa 10381 gttcttttaa aagagaattt agtaaagccc taagtctttt aataaaaaat tttttgtaaa 10441 aaagttggca aaaatagttt ttgctatata tttatatata agaaaaaatt caatactcaa 10501 ggagataaaa aatggaaaac ttaaacaata atccacaaaa aattaatcaa ataaataagc 10561 aaactaattt tattggtttt gaaaatttta tagaacttga taatcaagtt aatcaaacta 10621 aagaaaaaag taaattaagt aagataggta agaaattacc cggaataagc agtcaagtct 10681 agtgcggccg gaaacagcta tgacatgatt acgaattcga gctccttgtt ttcaatgata 10741 ggttttttag gtagagactt gtaagggttt gttcctggaa tataattttt aggaattttc 10801 atatatttta ctcctattta actagctaac ctataattct attatttttt ttgtttttat 10861 gcaatcaata ttgatatttt gattttttat gattagaatc atcatggaag agccaaagtc 10921 aacacttata gacaaatatg atctttctaa ccttcacgaa atagcatcag ggggagcacc 10981 tttgtctaag gaagtaggag aggcagttgc aaagaggttt catttacccg gaattaggca 11041 aggatatggg ttaacagaaa ccacatctgc aattttaatt acacctgaag gagatgataa 11101 accaggagct gttgggaagg ttgttccttt ttttgaggct aaagttgttg atttagatac 11161 tggaaagact ttaggagtta atcaaagagg cgagttatgt gtaagaggac ctatgataat 11221 gtcaggatac gtaaataatc cagaagctac aaatgcattg atagataaag atggttggtt 11281 acactcaggc gatatagctt attgggacga agatgaacac ttttttatag ttgatagatt 11341 gaaatctttg attaagtata aaggttatca agtagctcca gcagaattgg aaagtatttt 11401 attgcaacac ccagctcttc cgttttagag ctagaaatag caagttaaaa taaggctagt 11461 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttgg //