LOCUS Exported 11504 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION pZ8-1 plasmid carrying dcas9 driven by the propionate-inducible prpD2 promoter (PprpD2), KanR. ACCESSION . VERSION . KEYWORDS pZ8-P_dCas9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11504) AUTHORS Cleto S, Jensen JV, Wendisch VF, Lu TK TITLE Corynebacterium glutamicum Metabolic Engineering with CRISPR Interference (CRISPRi). JOURNAL ACS Synth Biol. 2016 Feb 16. PUBMED 26829286 REFERENCE 2 (bases 1 to 11504) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11504 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 521..586 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 593..613 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 623..4726 /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" CDS 4727..4747 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(4833..4850) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 5003..5089 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 5181..5208 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 5227..5318 /gene="bla" /label=AmpR promoter primer_bind complement(5537..5556) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" CDS complement(9263..10078) /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 TAFQVLEEYPDSGENIADALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 9986..10005 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 10070..10089 /label=pENTR-R /note="pENTR vectors, reverse primer" primer_bind complement(10539..10556) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(10673..11218) /direction=LEFT /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. " primer_bind complement(10753..10772) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cggtacgcct gcggccaaat ttgattcctt tcagctttgc ttcctgtcgg ccctcattcg 61 tgcgctctag gatcagtgat ccgggcttat cgactgcacg gtgcaccaat gcttctggcg 121 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg 181 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt 241 ctggcaaata ttctgaaatg agctgctcca gcgtccaaga atatgccccc gcgcgccggg 301 tggggagcga agggaacccc caaggaattg gcgttgaggt ggtgattttg catgttttac 361 tcaaaatcac tttgatggtc acaaaattac acaactttta cagtgaccta cattgctttt 421 taaagaatta gtgtggtgtg cgatccccgg gtacccagct tgagtaggac aaatccgccg 481 agcttcgacg agattttcag gagctaagga agctaaaatg gactacaaag accatgacgg 541 tgattataaa gatcatgaca tcgattacaa ggatgacgat gacaagatgg cccccaagaa 601 gaagaggaag gtgggccgcg gaatggataa gaaatactca ataggcttag ctatcggcac 661 aaatagcgtc ggatgggcgg tgatcactga tgaatataag gttccgtcta aaaagttcaa 721 ggttctggga aatacagacc gccacagtat caaaaaaaat cttatagggg ctcttttatt 781 tgacagtgga gagacagcgg aagcgactcg tctcaaacgg acagctcgta gaaggtatac 841 acgtcggaag aatcgtattt gttatctaca ggagattttt tcaaatgaga tggcgaaagt 901 agatgatagt ttctttcatc gacttgaaga gtcttttttg gtggaagaag acaagaagca 961 tgaacgtcat cctatttttg gaaatatagt agatgaagtt gcttatcatg agaaatatcc 1021 aactatctat catctgcgaa aaaaattggt agattctact gataaagcgg atttgcgctt 1081 aatctatttg gccttagcgc atatgattaa gtttcgtggt cattttttga ttgagggaga 1141 tttaaatcct gataatagtg atgtggacaa actatttatc cagttggtac aaacctacaa 1201 tcaattattt gaagaaaacc ctattaacgc aagtggagta gatgctaaag cgattctttc 1261 tgcacgattg agtaaatcaa gacgattaga aaatctcatt gctcagctcc ccggtgagaa 1321 gaaaaatggc ttatttggga atctcattgc tttgtcattg ggtttgaccc ctaattttaa 1381 atcaaatttt gatttggcag aagatgctaa attacagctt tcaaaagata cttacgatga 1441 tgatttagat aatttattgg cgcaaattgg agatcaatat gctgatttgt ttttggcagc 1501 taagaattta tcagatgcta ttttactttc agatatccta agagtaaata ctgaaataac 1561 taaggctccc ctatcagctt caatgattaa acgctacgat gaacatcatc aagacttgac 1621 tcttttaaaa gctttagttc gacaacaact tccagaaaag tataaagaaa tcttttttga 1681 tcaatcaaaa aacggatatg caggttatat tgatggggga gctagccaag aagaatttta 1741 taaatttatc aaaccaattt tagaaaaaat ggatggtact gaggaattat tggtgaaact 1801 aaatcgtgaa gatttgctgc gcaagcaacg gacctttgac aacggctcta ttccccatca 1861 aattcacttg ggtgagctgc atgctatttt gagaagacaa gaagactttt atccattttt 1921 aaaagacaat cgtgagaaga ttgaaaaaat cttgactttt cgaattcctt attatgttgg 1981 tccattggcg cgtggcaata gtcgttttgc atggatgact cggaagtctg aagaaacaat 2041 taccccatgg aattttgaag aagttgtcga taaaggtgct tcagctcaat catttattga 2101 acgcatgaca aactttgata aaaatcttcc aaatgaaaaa gtactaccaa aacatagttt 2161 gctttatgag tattttacgg tttataacga attgacaaag gtcaaatatg ttactgaagg 2221 aatgcgaaaa ccagcatttc tttcaggtga acagaagaaa gccattgttg atttactctt 2281 caaaacaaat cgaaaagtaa ccgttaagca attaaaagaa gattatttca aaaaaataga 2341 atgttttgat agtgttgaaa tttcaggagt tgaagataga tttaatgctt cattaggtac 2401 ctaccatgat ttgctaaaaa ttattaaaga taaagatttt ttggataatg aagaaaatga 2461 agatatctta gaggatattg ttttaacatt gaccttattt gaagataggg agatgattga 2521 ggaaagactt aaaacatatg ctcacctctt tgatgataag gtgatgaaac agcttaaacg 2581 tcgccgttat actggttggg gacgtttgtc tcgaaaattg attaatggta ttagggataa 2641 gcaatctggc aaaacaatat tagatttttt gaaatcagat ggttttgcca atcgcaattt 2701 tatgcagctg atccatgatg atagtttgac atttaaagaa gacattcaaa aagcacaagt 2761 gtctggacaa ggcgatagtt tacatgaaca tattgcaaat ttagctggta gccctgctat 2821 taaaaaaggt attttacaga ctgtaaaagt tgttgatgaa ttggtcaaag taatggggcg 2881 gcataagcca gaaaatatcg ttattgaaat ggcacgtgaa aatcagacaa ctcaaaaggg 2941 ccagaaaaat tcgcgagagc gtatgaaacg aatcgaagaa ggtatcaaag aattaggaag 3001 tcagattctt aaagagcatc ctgttgaaaa tactcaattg caaaatgaaa agctctatct 3061 ctattatctc caaaatggaa gagacatgta tgtggaccaa gaattagata ttaatcgttt 3121 aagtgattat gatgtcgatc acattgttcc acaaagtttc cttaaagacg attcaataga 3181 caataaggtc ttaacgcgtt ctgataaaaa tcgtggtaaa tcggataacg ttccaagtga 3241 agaagtagtc aaaaagatga aaaactattg gagacaactt ctaaacgcca agttaatcac 3301 tcaacgtaag tttgataatt taacgaaagc tgaacgtgga ggtttgagtg aacttgataa 3361 agctggtttt atcaaacgcc aattggttga aactcgccaa atcactaagc atgtggcaca 3421 aattttggat agtcgcatga atactaaata cgatgaaaat gataaactta ttcgagaggt 3481 taaagtgatt accttaaaat ctaaattagt ttctgacttc cgaaaagatt tccaattcta 3541 taaagtacgt gagattaaca attaccatca tgcccatgat gcgtatctaa atgccgtcgt 3601 tggaactgct ttgattaaga aatatccaaa acttgaatcg gagtttgtct atggtgatta 3661 taaagtttat gatgttcgta aaatgattgc taagtctgag caagaaatag gcaaagcaac 3721 cgcaaaatat ttcttttact ctaatatcat gaacttcttc aaaacagaaa ttacacttgc 3781 aaatggagag attcgcaaac gccctctaat cgaaactaat ggggaaactg gagaaattgt 3841 ctgggataaa gggcgagatt ttgccacagt gcgcaaagta ttgtccatgc cccaagtcaa 3901 tattgtcaag aaaacagaag tacagacagg cggattctcc aaggagtcaa ttttaccaaa 3961 aagaaattcg gacaagctta ttgctcgtaa aaaagactgg gatccaaaaa aatatggtgg 4021 ttttgatagt ccaacggtag cttattcagt cctagtggtt gctaaggtgg aaaaagggaa 4081 atcgaagaag ttaaaatccg ttaaagagtt actagggatc acaattatgg aaagaagttc 4141 ctttgaaaaa aatccgattg actttttaga agctaaagga tataaggaag ttaaaaaaga 4201 cttaatcatt aaactaccta aatatagtct ttttgagtta gaaaacggtc gtaaacggat 4261 gctggctagt gccggagaat tacaaaaagg aaatgagctg gctctgccaa gcaaatatgt 4321 gaatttttta tatttagcta gtcattatga aaagttgaag ggtagtccag aagataacga 4381 acaaaaacaa ttgtttgtgg agcagcataa gcattattta gatgagatta ttgagcaaat 4441 cagtgaattt tctaagcgtg ttattttagc agatgccaat ttagataaag ttcttagtgc 4501 atataacaaa catagagaca aaccaatacg tgaacaagca gaaaatatta ttcatttatt 4561 tacgttgacg aatcttggag ctcccgctgc ttttaaatat tttgatacaa caattgatcg 4621 taaacgatat acgtctacaa aagaagtttt agatgccact cttatccatc aatccatcac 4681 tggtctttat gaaacacgca ttgatttgag tcagctagga ggtgacccca agaagaagag 4741 gaaggtgggc tgacacacag gaaacagaat tcccggggat ccgtcgacct gcagccaagc 4801 ttctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga 4861 agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc 4921 atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg 4981 agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt 5041 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc 5101 ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac 5161 tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca 5221 aactcttttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 5281 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg 5341 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc 5401 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg 5461 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga 5521 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc 5581 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactgc ggcgtcgctg 5641 atcgccctgg cgacgttgtg cgggtggctt gtccctgagg gcgctgcgac agatagctaa 5701 aaatctgggt caggatcgcc gtagagcgcg cgtcgtcgat tggaggcttc ccctttggtt 5761 gacggtcttc aatcgctcta cggcgatcct gacgcttttt tgttgcgtac cgtcgatcgt 5821 tttatttctg tcgatcccga aaaagttttt gccttttgta aaaaacttct cggtcgcccc 5881 gcaaattttc gattccagat tttttaaaaa ccaagccaga aatacgacac accgtttgca 5941 gataatctgt ctttcggaaa aatcaagtgc gatacaaaat ttttagcacc cctgagctgc 6001 gcaaagtccc gcttcgtgaa aattttcgtg ccgcgtgatt ttccgccaaa aactttaacg 6061 aacgttcgtt ataatggtgt catgaccttc acgacgaagt actaaaattg gcccgaatca 6121 tcagctatgg atctctctga tgtcgcgctg gagtccgacg cgctcgatgc tgccgtcgat 6181 ttaaaaacgg tgatcggatt tttccgagct ctcgatacga cggacgcgcc agcatcacga 6241 gactgggcca gtgccgcgag cgacctagaa actctcgtgg cggatcttga ggagctggct 6301 gacgagctgc gtgctcggcc agcgccagga ggacgcacag tagtggagga tgcaatcagt 6361 tgcgcctact gcggtggcct gattcctccc cggcctgacc cgcgaggacg gcgcgcaaaa 6421 tattgctcag atgcgtgtcg tgccgcagcc agccgcgagc gcgccaacaa acgccacgcc 6481 gaggagctgg aggcggctag gtcgcaaatg gcgctggaag tgcgtccccc gagcgaaatt 6541 ttggccatgg tcgtcacaga gctggaagcg gcagcgagaa ttatccgcga tcgtggcgcg 6601 gtgcccgcag gcatgacaaa catcgtaaat gccgcgtttc gtgtggccgt ggccgcccag 6661 gacgtgtcag cgccgccacc acctgcaccg aatcggcagc agcgtcgcgc gtcgaaaaag 6721 cgcacaggcg gcaagaagcg ataagctgca cgaatacctg aaaaatgttg aacgccccgt 6781 gagcggtaac tcacagggcg tcggctaacc cccagtccaa acctgggaga aagcgctcaa 6841 aaatgactct agcggattca cgagacattg acacaccggc ctggaaattt tccgctgatc 6901 tgttcgacac ccatcccgag ctcgcgctgc gatcacgtgg ctggacgagc gaagaccgcc 6961 gcaaattcct cgctcacctg ggcagagaaa atttccaggg cagcaagacc cgcgacttcg 7021 ccagcgcttg gatcaaagac ccggacacgg gagaaacaca gccgaagtta taccgagttg 7081 gttcaaaatc gcttgcccgg tgccagtatg ttgctctgac gcacgcgcag cacgcagccg 7141 tgcttgtcct ggacattgat gtgccgagcc accaggccgg cgggaaaatc gagcacgtaa 7201 accccgaggt ctacgcgatt ttggagcgct gggcacgcct ggaaaaagcg ccagcttgga 7261 tcggcgtgaa tccactgagc gggaaatgcc agctcatctg gctcattgat ccggtgtatg 7321 ccgcagcagg catgagcagc ccgaatatgc gcctgctggc tgcaacgacc gaggaaatga 7381 cccgcgtttt cggcgctgac caggcttttt cacataggct gagccgtggc cactgcactc 7441 tccgacgatc ccagccgtac cgctggcatg cccagcacaa tcgcgtggat cgcctagctg 7501 atcttatgga ggttgctcgc atgatctcag gcacagaaaa acctaaaaaa cgctatgagc 7561 aggagttttc tagcggacgg gcacgtatcg aagcggcaag aaaagccact gcggaagcaa 7621 aagcacttgc cacgcttgaa gcaagcctgc cgagcgccgc tgaagcgtct ggagagctga 7681 tcgacggcgt ccgtgtcctc tggactgctc cagggcgtgc cgcccgtgat gagacggctt 7741 ttcgccacgc tttgactgtg ggataccagt taaaagcggc tggtgagcgc ctaaaagaca 7801 ccaagggtca tcgagcctac gagcgtgcct acaccgtcgc tcaggcggtc ggaggaggcc 7861 gtgagcctga tctgccgccg gactgtgacc gccagacgga ttggccgcga cgtgtgcgcg 7921 gctacgtcgc taaaggccag ccagtcgtcc ctgctcgtca gacagagacg cagagccagc 7981 cgaggcgaaa agctctggcc actatgggaa gacgtggcgg taaaaaggcc gcagaacgct 8041 ggaaagaccc aaacagtgag tacgcccgag cacagcgaga aaaactagct aagtccagtc 8101 aacgacaagc taggaaagct aaaggaaatc gcttgaccat tgcaggttgg tttatgactg 8161 ttgagggaga gactggctcg tggccgacaa tcaatgaagc tatgtctgaa tttagcgtgt 8221 cacgtcagac cgtgaataga gcacttaagg tctgcgggca ttgaacttcc acgaggacgc 8281 cgaaagcttc ccagtaaatg tgccatctcg taggcagaaa acggttcccc cgtagggtct 8341 ctctcttggc ctcctttcta ggtcgggctg attgctcttg aagctctcta ggggggctca 8401 caccataggc agataacgtt ccccaccggc tcgcctcgta agcgcacaag gactgctccc 8461 aaagatcttc aaagccactg ccgcgactgc cttcgcgaag ccttgccccg cggaaatttc 8521 ctccaccgag ttcgtgcaca cccctatgcc aagcttcttt caccctaaat tcgagagatt 8581 ggattcttac cgtggaaatt cttcgcaaaa atcgtcccct gatcgccctt gcgacgttgg 8641 cgtcggtgcc gctggttgcg cttggcttga ccgacttgat cctccggcgt tcagcctgtg 8701 ccacagccga caggatggtg accaccattt gccccatatc accgtcggta ctgatcccgt 8761 cgtcaataaa ccgaaccgct acaccctgag catcaaactc ttttatcagt tggatcatgt 8821 cggcgtgtcg cggccaagac ggtcgagctt cttcaccaga atgacatcac cttcctccac 8881 cttcatcctc agcaaatcca gcccttcccg atctgttgaa ctgccggatg ccttgtcggt 8941 aaagatgcgg ttagctttta cccctgcatc tttgagcgct gaggtctgcc tcgtgaagaa 9001 ggtgttgctg actcatacca ggcctgaatc gccccatcat ccagccagaa agtgagggag 9061 ccacggttga tgagagcttt gttgtaggtg gaccagttgg tgattttgaa cttttgcttt 9121 gccacggaac ggtctgcgtt gtcgggaaga tgcgtgatct gatccttcaa ctcagcaaaa 9181 gttcgattta ttcaacaaag ccgccgtccc gtcaagtcag cgtaatgctc tgccagtgtt 9241 acaaccaatt aaccaattct gattagaaaa actcatcgag catcaaatga aactgcaatt 9301 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag 9361 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga 9421 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg 9481 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagctta tgcatttctt 9541 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca 9601 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag 9661 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcagcaa 9721 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg 9781 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag 9841 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc 9901 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga 9961 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat 10021 ccatgttgga atttaatcgc ggcctcgagc aagacgtttc ccgttgaata tggctcataa 10081 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt 10141 tatcttgtgc aatgtaacat cagagatttt gagacacaac gtggctttgt tgaataaatc 10201 gaacttttgc tgagttgaag gatcagatca cgcatcttcc cgacaacgca gaccgttccg 10261 tggcaaagca aaagttcaaa atcaccaact ggtccaccta caacaaagct ctcatcaacc 10321 gtggctccct cactttctgg ctggatgatg gggcgattca ggcctggtat gagtcagcaa 10381 caccttcttc acgaggcaga cctcagcgct agcggagtgt atactggctt actatgttgg 10441 cactgatgag ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg 10501 cgtcagcaga atatgtgata caggatatat tccgcttcct cgctcactga ctcgctacgc 10561 tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga tttcctggaa 10621 gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg tttttccata 10681 ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc 10741 cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg cgctctcctg 10801 ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg tctcattcca 10861 cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat gcacgaaccc 10921 cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggaa 10981 agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg agttagtctt 11041 gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact gcgctcctcc 11101 aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa aaccgccctg 11161 caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac gatctcaaga 11221 agatcatctt attaaggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 11281 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 11341 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 11401 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 11461 cgtgtagata actacgatac gggagggctt accatctggc ccca //