LOCUS Exported 15876 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Cas9 only expression, Cas9 Type: TaCas9_D10A+H840A (dead), Plant Selection: PvUbi2:bar. ACCESSION . VERSION . KEYWORDS pTaCas9-dead_6 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 15876) AUTHORS Cermak T, Curtin SJ, Gil-Humanes J, Cegan R, Kono TJY, Konecna E, Belanto JJ, Starker CG, Mathre JW, Greenstein RL, Voytas DF TITLE A multi-purpose toolkit to enable advanced genome engineering in plants. JOURNAL Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. PUBMED 28522548 REFERENCE 2 (bases 1 to 15876) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..15876 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(291..1085) /codon_start=1 /gene="aphA-3" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF" misc_feature 1510..1534 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" polyA_signal 1612..1786 /label=CaMV poly(A) signal /note="cauliflower mosaic virus polyadenylation signal" CDS complement(1793..2344) /codon_start=1 /gene="Streptomyces hygroscopicus bar" /product="phosphinothricin acetyltransferase" /label=BlpR /note="confers resistance to bialophos or phosphinothricin" /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF WQLDFSLPVPPRPVLPVTEI" protein_bind 4284..4305 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4320..4350 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4358..4374 /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 4363..4385 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 4382..4398 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4382..4398 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 6455..10558 /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 TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 10559..10606 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" primer_bind complement(10887..10904) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(10887..10903) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(10896..10918) /label=M13/pUC Forward /note="In lacZ gene" misc_feature 11106..11130 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS 12431..13060 /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" CDS 13489..14562 /codon_start=1 /product="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 RepA /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI LVMRYRNLIEGEASAGS" rep_origin 14628..14822 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" primer_bind complement(15058..15080) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 15166..15306 /label=bom /note="basis of mobility region from pBR322" primer_bind 15221..15240 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(15321..15338) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(15492..204) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(15572..15591) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca 61 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 121 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 181 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 241 cagtggaacg aaaactcacg ttaagggatt ttggtcatgc attctaggta ctaaaacaat 301 tcatccagta aaatataata ttttattttc tcccaatcag gcttgatccc cagtaagtca 361 aaaaatagct cgacatactg ttcttccccg atatcctccc tgatcgaccg gacgcagaag 421 gcaatgtcat accacttgtc cgccctgccg cttctcccaa gatcaataaa gccacttact 481 ttgccatctt tcacaaagat gttgctgtct cccaggtcgc cgtgggaaaa gacaagttcc 541 tcttcgggct tttccgtctt taaaaaatca tacagctcgc gcggatcttt aaatggagtg 601 tcttcttccc agttttcgca atccacatcg gccagatcgt tattcagtaa gtaatccaat 661 tcggctaagc ggctgtctaa gctattcgta tagggacaat ccgatatgtc gatggagtga 721 aagagcctga tgcactccgc atacagctcg ataatctttt cagggctttg ttcatcttca 781 tactcttccg agcaaaggac gccatcggcc tcactcatga gcagattgct ccagccatca 841 tgccgttcaa agtgcaggac ctttggaaca ggcagctttc cttccagcca tagcatcatg 901 tccttttccc gttccacatc ataggtggtc cctttatacc ggctgtccgt catttttaaa 961 tataggtttt cattttctcc caccagctta tataccttag caggagacat tccttccgta 1021 tcttttacgc agcggtattt ttcgatcagt tttttcaatt ccggtgatat tctcatttta 1081 gccatttatt atttccttcc tcttttctac agtatttaaa gataccccaa gaagctaatt 1141 ataacaagac gaactccaat tcactgttcc ttgcattcta aaaccttaaa taccagaaaa 1201 cagctttttc aaagttgttt tcaaagttgg cgtataacat agtatcgacg gagccgattt 1261 tgaaaccgcg gtgatcacag gcagcaacgc tctgtcatcg ttacaatcaa catgctaccc 1321 tccgcgagat catccgtgtt tcaaacccgg cagcttagtt gccgttcttc cgaatagcat 1381 cggtaacatg agcaaagtct gccgccttac aacggctctc ccgctgacgc cgtcccggac 1441 tgatgggctg cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg 1501 gctggctggt ggcaggatat attgtggtgt aaacaaattg acgcttagac aacttaataa 1561 cacattgcgg acgtttttaa tgtactgaat taacgccgaa ttaattcggg ggatctggat 1621 tttagtactg gattttggtt ttaggaatta gaaattttat tgatagaagt attttacaaa 1681 tacaaataca tactaagggt ttcttatatg ctcaacacat gagcgaaacc ctataggaac 1741 cctaattccc ttatctggga actactcaca cattattatg gagaaactcg agtcaaatct 1801 cggtgacggg caggaccgga cggggcggta ccggcaggct gaagtccagc tgccagaaac 1861 ccacgtcatg ccagttcccg tgcttgaagc cggccgcccg cagcatgccg cggggggcat 1921 atccgagcgc ctcgtgcatg cgcacgctcg ggtcgttggg cagcccgatg acagcgacca 1981 cgctcttgaa gccctgtgcc tccagggact tcagcagatg ggtgtagagc gtggagccca 2041 gtcccgtccg ctggtggcgg gggctgacgt acacggtgga ctcggccgtc cagtcgtagg 2101 cgttgcgtgc cttccaggga cccgcgtagg cgatgccggc gacctcgccg tccacctcgg 2161 cgacgagcca gggatagcgc tcccgcagac ggacgaggtc gtccgtccac tcctgcggtt 2221 cctgcggctc ggtacggaag ttgaccgtgc ttgtctcgat gtagtggttg acgatggtgc 2281 agaccgccgg catgtccgcc tcggtggcac ggcggatgtc ggccgggcgt cgttctgggc 2341 tcataagctt ctgcaaaaga gaaccagaca acagggtaag tgcctagcag taaacaaaca 2401 gaactcatca caagcaaaca gcaacatcat attcatacca acaggtcatg tgtgttcatc 2461 acatcattag tactaagcat gccatcatcc aagtatatca aagtaagggc aaagagcatg 2521 catgatcatc aggtgcacaa aagaatcatc aaattgtagc agtacatatc ttcatctatc 2581 atgcatatct atccataaca ggacgatgca tgttgaccag gtaaaagcta caggatccta 2641 taggaacagc agcgtatata tcttcaccaa tcttagcatc ttaatcatgt ggcacatgca 2701 gtttcaattt aagcacatga gctagttgat tatgaggtac cagagaatca tcaaactgat 2761 gtagcagcat atatcctcat ctatcatgca atctaatcta atctaactaa acaggaaagg 2821 tgtgctattc agttaaaagc taccgcatca tatacaaacg gcagcataag aaaaagcata 2881 atcatcttaa tcatgcaaca aacgcagatt cataataagc ccaagagcta gcttgtgatg 2941 atcttattct actctgatct acagcaatca gataacgacc taaccttgca catggcaaca 3001 aaacaatcga tcggacgaat cagttgtttg ttcctagcta gcaccatcga accagataat 3061 agatgcacgt acagatcccg aaaacgaacc caaaaacagg gcagacctag ctgaacctag 3121 gcagcgaccc agcagatcgt gagaacgatc tcatctacga acagcctaga agcaacccca 3181 cgattcccgg acaaacgacc taaaatcccc acaaatcaca tgagcatgac aggataaaca 3241 gcggaaccga tcagatctac acgaaaaccc cacctcccag ccacccacga tcaggaaaca 3301 cgcggatcta gcatgatttc gtcaacgcct cagcctagtt cctagccaca gaccaagcag 3361 aaccaccaaa ccacgccgag cgaggagatg gggcaagagg acgggggaga cgatcgccgt 3421 accttgaagc gggggaagga tcgccgaggg tcgcgaggag agcaatttgg atttggaacc 3481 cgggggttgt gcgctccgaa cgatgagacg atgtgagatt gtgggaagag gcgcggaggg 3541 ccctgtattt atgggctgcg acggggggag gagaggtggg gagggttggg gaaggaatcc 3601 cccacccgtg ccgtgacggt tccgggccgt gtgagaggag cccgctcgtc tccgccacgc 3661 aatttccgcg atcggagcgg agctttcgag aggcggctgg atggttggtg gccgttagat 3721 ttgtagacgc cgttaacgcc tcgcctccac cgggaagagt tttgagcagc cgcttatgac 3781 aatggcttaa cgacgttaga cggagcgtta gtggcaggcc atgatagaat agacgtattg 3841 caatgggatt ataattaaat aaataagaat ataataagat atggcaagtc ggcactcatg 3901 acatggtctt cgaaatgata gtgctcactt tcttagccga gaaagttgac gcgactgatt 3961 tagaagttaa gatattattt ctctcttctt ttctttcctc gtcatataag gatgaaataa 4021 actttagaga ttgccggtag tgattttgga tttcggcgat caggcttggt ttgccggttt 4081 cggacggtgt gccttaggcc acccgcagtg tatcttgtaa tgttcaaccg ataagcaagg 4141 gtggggctca agcaagtagt aaacaactat gtcaaatgtc accatggtta tggtcttgtt 4201 tagttggctt cttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa 4261 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct 4321 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac 4381 acaggaaaca gctatgacat gattacgaat tcgagctcgg tacccgggga tctgcaggcg 4441 cgcctgcagt gcagcgtgac ccggtcgtgc ccctctctag agataatgag cattgcatgt 4501 ctaagttata aaaaattacc acatattttt tttgtcacac ttgtttgaag tgcagtttat 4561 ctatctttat acatatattt aaactttact ctacgaataa tataatctat agtactacaa 4621 taatatcagt gttttagaga atcatataaa tgaacagtta gacatggtct aaaggacaat 4681 tgagtatttt gacaacagga ctctacagtt ttatcttttt agtgtgcatg tgttctcctt 4741 tttttttgca aatagcttca cctatataat acttcatcca ttttattagt acatccattt 4801 agggtttagg gttaatggtt tttatagact aattttttta gtacatctat tttattctat 4861 tttagcctct aaattaagaa aactaaaact ctattttagt ttttttattt aataatttag 4921 atataaaata gaataaaata aagtgactaa aaattaaaca aatacccttt aagaaattaa 4981 aaaaactaag gaaacatttt tcttgtttcg agtagataat gccagcctgt taaacgccgt 5041 cgacgagtct aacggacacc aaccagcgaa ccagcagcgt cgcgtcgggc caagcgaagc 5101 agacggcacg gcatctctgt cgctgcctct ggacccctct cgagagttcc gctccaccgt 5161 tggacttgct ccgctgtcgg catccagaaa ttgcgtggcg gagcggcaga cgtgagccgg 5221 cacggcaggc ggcctcctcc tcctctcacg gcaccggcag ctacggggga ttcctttccc 5281 accgctcctt cgctttccct tcctcgcccg ccgtaataaa tagacacccc ctccacaccc 5341 tctttcccca acctcgtgtt gttcggagcg cacacacaca caaccagatc tcccccaaat 5401 ccacccgtcg gcacctccgc ttcaaggtac gccgctcgtc ctcccccccc cccctctcta 5461 ccttctctag atcggcgttc cggtccatgg ttagggcccg gtagttctac ttctgttcat 5521 gtttgtgtta gatccgtgtt tgtgttagat ccgtgctgct agcgttcgta cacggatgcg 5581 acctgtacgt cagacacgtt ctgattgcta acttgccagt gtttctcttt ggggaatcct 5641 gggatggctc tagccgttcc gcagacggga tcgatttcat gatttttttt gtttcgttgc 5701 atagggtttg gtttgccctt ttcctttatt tcaatatatg ccgtgcactt gtttgtcggg 5761 tcatcttttc atgctttttt ttgtcttggt tgtgatgatg tggtctggtt gggcggtcgt 5821 tctagatcgg agtagaatta attctgtttc aaactacctg gtggatttat taattttgga 5881 tctgtatgtg tgtgccatac atattcatag ttacgaattg aagatgatgg atggaaatat 5941 cgatctagga taggtataca tgttgatgcg ggttttactg atgcatatac agagatgctt 6001 tttgttcgct tggttgtgat gatgtggtgt ggttgggcgg tcgttcattc gttctagatc 6061 ggagtagaat actgtttcaa actacctggt gtatttatta attttggaac tgtatgtgtg 6121 tgtcatacat cttcatagtt acgagtttaa gatggatgga aatatcgatc taggataggt 6181 atacatgttg atgtgggttt tactgatgca tatacatgat ggcatatgca gcatctattc 6241 atatgctcta accttgagta cctatctatt ataataaaca agtatgtttt ataattattt 6301 tgatcttgat atacttggat gatggcatat gcagcagcta tatgtggatt tttttagccc 6361 tgccttcata cgctatttat ttgcttggta ctgtttcttt tgtcgatgct caccctgttg 6421 tttggtgtta cttcctgcag ggcgatctat tcgaatggac aagaagtact cgatcggcct 6481 cgccatcggg acgaactcag ttggctgggc cgtgatcacc gacgagtaca aggtgccctc 6541 taagaagttc aaggtcctgg ggaacaccga ccgccattcc atcaagaaga acctcatcgg 6601 cgctctcctg ttcgacagcg gggagaccgc tgaggctacg aggctcaaga gaaccgctag 6661 gcgccggtac acgagaagga agaacaggat ctgctacctc caagagattt tctccaacga 6721 gatggccaag gttgacgatt cattcttcca ccgcctggag gagtctttcc tcgtggagga 6781 ggataagaag cacgagcggc atcccatctt cggcaacatc gtggacgagg ttgcctacca 6841 cgagaagtac cctacgatct accatctgcg gaagaagctc gtggactcca ccgataaggc 6901 ggacctcaga ctgatctacc tcgctctggc ccacatgatc aagttccgcg gccatttcct 6961 gatcgagggg gatctcaacc cagacaacag cgatgttgac aagctgttca tccaactcgt 7021 gcagacctac aaccaactct tcgaggagaa cccgatcaac gcctctggcg tggacgcgaa 7081 ggctatcctg tccgcgaggc tctcgaagtc caggaggctg gagaacctga tcgctcagct 7141 cccaggcgag aagaagaacg gcctgttcgg gaacctcatc gctctcagcc tggggctcac 7201 cccgaacttc aagtcgaact tcgatctcgc tgaggacgcc aagctgcaac tctccaagga 7261 cacctacgac gatgacctcg ataacctcct ggcccagatc ggcgatcaat acgcggacct 7321 gttcctcgct gccaagaacc tgtcggacgc catcctcctg tcagatatcc tccgcgtgaa 7381 caccgagatc acgaaggctc cactctctgc ctccatgatc aagcgctacg acgagcacca 7441 tcaggatctg accctcctga aggcgctggt ccgccaacag ctcccggaga agtacaagga 7501 gattttcttc gatcagtcga agaacggcta cgctgggtac atcgacggcg gggcctcaca 7561 agaggagttc tacaagttca tcaagccaat cctggagaag atggacggca cggaggagct 7621 cctggtgaag ctcaacaggg aggacctcct gcggaagcag agaaccttcg ataacggcag 7681 catcccccac caaatccatc tcggggagct gcacgccatc ctgagaaggc aagaggactt 7741 ctaccctttc ctcaaggata accgggagaa gatcgagaag atcctgacct tcagaatccc 7801 atactacgtc ggccctctcg cgcgggggaa ctcaagattc gcttggatga cccgcaagtc 7861 tgaggagacc atcacgccgt ggaacttcga ggaggtggtg gacaagggcg ctagcgctca 7921 gtcgttcatc gagaggatga ccaacttcga caagaacctg cccaacgaga aggtgctccc 7981 taagcactcg ctcctgtacg agtacttcac cgtctacaac gagctcacga aggtgaagta 8041 cgtcaccgag ggcatgcgca agccagcgtt cctgtccggg gagcagaaga aggctatcgt 8101 ggacctcctg ttcaagacca accggaaggt cacggttaag caactcaagg aggactactt 8161 caagaagatc gagtgcttcg attcggtcga gatcagcggc gttgaggacc gcttcaacgc 8221 cagcctcggg acctaccacg atctcctgaa gatcatcaag gataaggact tcctggacaa 8281 cgaggagaac gaggatatcc tggaggacat cgtgctgacc ctcacgctgt tcgaggacag 8341 ggagatgatc gaggagcgcc tgaagacgta cgcccatctc ttcgatgaca aggtcatgaa 8401 gcaactcaag cgccggagat acaccggctg ggggaggctg tcccgcaagc tcatcaacgg 8461 catccgggac aagcagtccg ggaagaccat cctcgacttc ctgaagagcg atggcttcgc 8521 caacaggaac ttcatgcaac tgatccacga tgacagcctc accttcaagg aggatatcca 8581 aaaggctcaa gtgagcggcc agggggactc gctgcacgag catatcgcga acctcgctgg 8641 ctcccccgcg atcaagaagg gcatcctcca gaccgtgaag gttgtggacg agctcgtgaa 8701 ggtcatgggc cggcacaagc ctgagaacat cgtcatcgag atggccagag agaaccaaac 8761 cacgcagaag gggcaaaaga actctaggga gcgcatgaag cgcatcgagg agggcatcaa 8821 ggagctgggg tcccaaatcc tcaaggagca cccagtggag aacacccaac tgcagaacga 8881 gaagctctac ctgtactacc tccagaacgg cagggatatg tacgtggacc aagagctgga 8941 tatcaaccgc ctcagcgatt acgacgtcga tgctatcgtt ccccagtctt tcctgaagga 9001 tgactccatc gacaacaagg tcctcaccag gtcggacaag aaccgcggca agtcagataa 9061 cgttccatct gaggaggtcg ttaagaagat gaagaactac tggaggcagc tcctgaacgc 9121 caagctgatc acgcaaagga agttcgacaa cctcaccaag gctgagagag gcgggctctc 9181 agagctggac aaggccggct tcatcaagcg gcagctggtc gagaccagac aaatcacgaa 9241 gcacgttgcg caaatcctcg actctcggat gaacacgaag tacgatgaga acgacaagct 9301 gatcagggag gttaaggtga tcaccctgaa gtctaagctc gtctccgact tcaggaagga 9361 tttccagttc tacaaggttc gcgagatcaa caactaccac catgcccatg acgcttacct 9421 caacgctgtg gtcggcaccg ctctgatcaa gaagtaccca aagctggagt ccgagttcgt 9481 gtacggggac tacaaggttt acgatgtgcg caagatgatc gccaagtcgg agcaagagat 9541 cggcaaggct accgccaagt acttcttcta ctcaaacatc atgaacttct tcaagaccga 9601 gatcacgctg gccaacggcg agatccggaa gagaccgctc atcgagacca acggcgagac 9661 gggggagatc gtgtgggaca agggcaggga tttcgcgacc gtccgcaagg ttctctccat 9721 gccccaggtg aacatcgtca agaagaccga ggtccaaacg ggcgggttct caaaggagtc 9781 tatcctgcct aagcggaaca gcgacaagct catcgccaga aagaaggact gggacccaaa 9841 gaagtacggc gggttcgaca gccctaccgt ggcctactcg gtcctggttg tggcgaaggt 9901 tgagaagggc aagtccaaga agctcaagag cgtgaaggag ctcctgggga tcaccatcat 9961 ggagaggtcc agcttcgaga agaacccaat cgacttcctg gaggccaagg gctacaagga 10021 ggtgaagaag gacctgatca tcaagctccc gaagtactct ctcttcgagc tggagaacgg 10081 caggaagaga atgctggctt ccgctggcga gctccagaag gggaacgagc tcgcgctgcc 10141 aagcaagtac gtgaacttcc tctacctggc ttcccactac gagaagctca agggcagccc 10201 ggaggacaac gagcaaaagc agctgttcgt cgagcagcac aagcattacc tcgacgagat 10261 catcgagcaa atctccgagt tcagcaagcg cgtgatcctc gccgacgcga acctggataa 10321 ggtcctctcc gcctacaaca agcaccggga caagcccatc agagagcaag cggagaacat 10381 catccatctc ttcaccctga cgaacctcgg cgctcctgct gctttcaagt acttcgacac 10441 cacgatcgat cggaagagat acacctccac gaaggaggtc ctggacgcga ccctcatcca 10501 ccagtcgatc accggcctgt acgagacgag gatcgacctc tcacaactcg gcggggataa 10561 gagacccgca gcaaccaaga aggcagggca agcaaagaag aagaagtgac tcgagatatg 10621 aagatgaaga tgaaatattt ggtgtgtcaa ataaaaagct tgtgtgctta agtttgtgtt 10681 tttttcttgg cttgttgtgt tatgaatttg tggctttttc taatattaaa tgaatgtaag 10741 atcacattat aatgaataaa caaatgtttc tataatccat tgtgaatgtt ttgttggatc 10801 tcttctgcag catataacta ctgtatgtgc tatggtatgg actatggaat atgattaaag 10861 ataaggagct ccggtgacgg actggcactg gccgtcgttt tacaacgtcg tgactgggaa 10921 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt 10981 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa 11041 tgctagagca gcttgagctt ggatcagatt gtcgtttccc gccttcagtt taaactatca 11101 gtgtttgaca ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataat 11161 cggatattta aaagggcgtg aaaaggttta tccgttcgtc catttgtatg tgcatgccaa 11221 ccacagggtt cccctcggga tcaaagtact ttgatccaac ccctccgctg ctatagtgca 11281 gtcggcttct gacgttcagt gcagccgtct tctgaaaacg acatgtcgca caagtcctaa 11341 gttacgcgac aggctgccgc cctgcccttt tcctggcgtt ttcttgtcgc gtgttttagt 11401 cgcataaagt agaatacttg cgactagaac cggagacatt acgccatgaa caagagcgcc 11461 gccgctggcc tgctgggcta tgcccgcgtc agcaccgacg accaggactt gaccaaccaa 11521 cgggccgaac tgcacgcggc cggctgcacc aagctgtttt ccgagaagat caccggcacc 11581 aggcgcgacc gcccggagct ggccaggatg cttgaccacc tacgccctgg cgacgttgtg 11641 acagtgacca ggctagaccg cctggcccgc agcacccgcg acctactgga cattgccgag 11701 cgcatccagg aggccggcgc gggcctgcgt agcctggcag agccgtgggc cgacaccacc 11761 acgccggccg gccgcatggt gttgaccgtg ttcgccggca ttgccgagtt cgagcgttcc 11821 ctaatcatcg accgcacccg gagcgggcgc gaggccgcca aggcccgagg cgtgaagttt 11881 ggcccccgcc ctaccctcac cccggcacag atcgcgcacg cccgcgagct gatcgaccag 11941 gaaggccgca ccgtgaaaga ggcggctgca ctgcttggcg tgcatcgctc gaccctgtac 12001 cgcgcacttg agcgcagcga ggaagtgacg cccaccgagg ccaggcggcg cggtgccttc 12061 cgtgaggacg cattgaccga ggccgacgcc ctggcggccg ccgagaatga acgccaagag 12121 gaacaagcat gaaaccgcac caggacggcc aggacgaacc gtttttcatt accgaagaga 12181 tcgaggcgga gatgatcgcg gccgggtacg tgttcgagcc gcccgcgcac ggctcaaccg 12241 tgcggctgca tgaaatcctg gccggtttgt ctgatgccaa gctggcggcc tggccggcca 12301 gcttggccgc tgaagaaacc gagcgccgcc gtctaaaaag gtgatgtgta tttgagtaaa 12361 acagcttgcg tcatgcggtc gctgcgtata tgatgcgatg agtaaataaa caaatacgca 12421 aggggaacgc atgaaggtta tcgctgtact taaccagaaa ggcgggtcag gcaagacgac 12481 catcgcaacc catctagccc gcgccctgca actcgccggg gccgatgttc tgttagtcga 12541 ttccgatccc cagggcagtg cccgcgattg ggcggccgtg cgggaagatc aaccgctaac 12601 cgttgtcggc atcgaccgcc cgacgattga ccgcgacgtg aaggccatcg gccggcgcga 12661 cttcgtagtg atcgacggag cgccccaggc ggcggacttg gctgtgtccg cgatcaaggc 12721 agccgacttc gtgctgattc cggtgcagcc aagcccttac gacatatggg ccaccgccga 12781 cctggtggag ctggttaagc agcgcattga ggtcacggat ggaaggctac aagcggcctt 12841 tgtcgtgtcg cgggcgatca aaggcacgcg catcggcggt gaggttgccg aggcgctggc 12901 cgggtacgag ctgcccattc ttgagtcccg tatcacgcag cgcgtgagct acccaggcac 12961 tgccgccgcc ggcacaaccg ttcttgaatc agaacccgag ggcgacgctg cccgcgaggt 13021 ccaggcgctg gccgctgaaa ttaaatcaaa actcatttga gttaatgagg taaagagaaa 13081 atgagcaaaa gcacaaacac gctaagtgcc ggccgtccga gcgcacgcag cagcaaggct 13141 gcaacgttgg ccagcctggc agacacgcca gccatgaagc gggtcaactt tcagttgccg 13201 gcggaggatc acaccaagct gaagatgtac gcggtacgcc aaggcaagac cattaccgag 13261 ctgctatctg aatacatcgc gcagctacca gagtaaatga gcaaatgaat aaatgagtag 13321 atgaatttta gcggctaaag gaggcggcat ggaaaatcaa gaacaaccag gcaccgacgc 13381 cgtggaatgc cccatgtgtg gaggaacggg cggttggcca ggcgtaagcg gctgggttgc 13441 ctgccggccc tgcaatggca ctggaacccc caagcccgag gaatcggcgt gagcggtcgc 13501 aaaccatccg gcccggtaca aatcggcgcg gcgctgggtg atgacctggt ggagaagttg 13561 aaggccgcgc aggccgccca gcggcaacgc atcgaggcag aagcacgccc cggtgaatcg 13621 tggcaagcgg ccgctgatcg aatccgcaaa gaatcccggc aaccgccggc agccggtgcg 13681 ccgtcgatta ggaagccgcc caagggcgac gagcaaccag attttttcgt tccgatgctc 13741 tatgacgtgg gcacccgcga tagtcgcagc atcatggacg tggccgtttt ccgtctgtcg 13801 aagcgtgacc gacgagctgg cgaggtgatc cgctacgagc ttccagacgg gcacgtagag 13861 gtttccgcag ggccggccgg catggccagt gtgtgggatt acgacctggt actgatggcg 13921 gtttcccatc taaccgaatc catgaaccga taccgggaag ggaagggaga caagcccggc 13981 cgcgtgttcc gtccacacgt tgcggacgta ctcaagttct gccggcgagc cgatggcgga 14041 aagcagaaag acgacctggt agaaacctgc attcggttaa acaccacgca cgttgccatg 14101 cagcgtacga agaaggccaa gaacggccgc ctggtgacgg tatccgaggg tgaagccttg 14161 attagccgct acaagatcgt aaagagcgaa accgggcggc cggagtacat cgagatcgag 14221 ctagctgatt ggatgtaccg cgagatcaca gaaggcaaga acccggacgt gctgacggtt 14281 caccccgatt actttttgat cgatcccggc atcggccgtt ttctctaccg cctggcacgc 14341 cgcgccgcag gcaaggcaga agccagatgg ttgttcaaga cgatctacga acgcagtggc 14401 agcgccggag agttcaagaa gttctgtttc accgtgcgca agctgatcgg gtcaaatgac 14461 ctgccggagt acgatttgaa ggaggaggcg gggcaggctg gcccgatcct agtcatgcgc 14521 taccgcaacc tgatcgaggg cgaagcatcc gccggttcct aatgtacgga gcagatgcta 14581 gggcaaattg ccctagcagg ggaaaaaggt cgaaaaggcc tctttcctgt ggatagcacg 14641 tacattggga acccaaagcc gtacattggg aaccggaacc cgtacattgg gaacccaaag 14701 ccgtacattg ggaaccggtc acacatgtaa gtgactgata taaaagagaa aaaaggcgat 14761 ttttccgcct aaaactcttt aaaacttatt aaaactctta aaacccgcct ggcctgtgca 14821 taactgtctg gccagcgcac agcccaagag ctgcaaaaag cgcctaccct tcggtcgctg 14881 cgctccctac gccccgccgc ttcgcgtcgg cctatcgcgg ccgctggccg ctcaaaaatg 14941 gctggcctac ggccaggcaa tctaccaggg cgcggacaag ccgcgccgtc gccactcgac 15001 cgccggcgcc cacatcaagg caccctgcct cgcgcgtttc ggtgatgacg gtgaaaacct 15061 ctgacacatg cagctcccgg aaacggtcac agcttgtctg taagcggatg ccgggagcag 15121 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca 15181 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta 15241 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc 15301 atcaggccct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 15361 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 15421 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 15481 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 15541 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 15601 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 15661 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 15721 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 15781 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 15841 gcagccactg gtaacaggat tagcagagcg aggtat //