LOCUS Exported 19102 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses R. toruloides codon-optimized SpCas9 under PGK1 promoter. ACCESSION . VERSION . KEYWORDS NM9-SpCas9-NLS3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 19102) AUTHORS Schultz JC, Cao M, Zhao H TITLE Development of a CRISPR/Cas9 system for high efficiency multiplexed gene deletion in Rhodosporidium toruloides. JOURNAL Biotechnol Bioeng. 2019 Apr 30. doi: 10.1002/bit.27001. PUBMED 31038202 REFERENCE 2 (bases 1 to 19102) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..19102 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 939..963 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" CDS complement(4092..6476) /codon_start=1 /product="C-terminal portion of Streptococcus pyogenes Cas9 (Zetsche et al., 2015)" /label=Cas9(C) /translation="CFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILED IVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGK TILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKG ILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQI LKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDN KVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDK AGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQF YKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGK ATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMP QVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKV EKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELEN GRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLD EIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKY FDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD" misc_feature 9930..9954 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS 11255..11884 /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" CDS 12313..13386 /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 13452..13646 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" primer_bind complement(13882..13904) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 13990..14130 /label=bom /note="basis of mobility region from pBR322" primer_bind 14045..14064 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(14145..14162) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(14316..14904) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(14396..14415) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(16329..17123) /codon_start=1 /gene="aphA-3" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF" CDS complement(17310..18113) /codon_start=1 /gene="S. cerevisiae URA3" /product="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /label=URA3 /note="yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGGDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" promoter complement(18114..18329) /gene="S. cerevisiae URA3" /label=URA3 promoter ORIGIN 1 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg 61 agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc 121 cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc 181 tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct 241 actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg 301 attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta 361 taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat 421 tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat 481 gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt 541 gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag 601 ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc 661 aaagagatac ttttgagcaa tgtttgtgga agcgccgcgg tgatcacagg cagcaacgct 721 ctgtcatcgt tacaatcaac atgctaccct ccgcgagatc atccgtgttt caaacccggc 781 agcttagttg ccgttcttcc gaatagcatc ggtaacatga gcaaagtctg ccgccttaca 841 acggctctcc cgctgacgcc gtcccggact gatgggctgc ctgtatcgag tggtgatttt 901 gtgccgagct gccggtcggg gagctgttgg ctggctggtg gcaggatata ttgtggtgta 961 aacaaattga cgcttagaca acttaataac acattgcgga cgtttttaat gtactgaatt 1021 aacgccgaat taattcctag gccaccatgt tgggcccggc gcgccgaatt cgagctcggt 1081 acccaactgc agaactacgc cctcgcgcat ccgacttcgg actcgaccgg cgaaacgact 1141 gcgacctatt tctactgctt gccttcgacg tccgggcgag tcgccgctta ccccgtgcgc 1201 tctcctcccc ctctggcccc tcaaactgca actcctgacc gcggcgatgc gcgcagtacc 1261 ccgtcaaaat gcaactctac gcgactttcg gcttagaagt cgccaagctc cgtgcttcgc 1321 cgcctcgagc tcttgcgttg cctgcagacg tcgtctatca cgaggcggag aagctcgagt 1381 tgccggcttt gccgagggcg gtcaaggttg agtcggagac tgtcaaggta gaagttcgag 1441 aagaggagga tgaggttaag ggtgaggtgc tgttagtgaa gacggactcg gttgtgcagg 1501 aggacactga agcgggctcg gaaggagtgc aggttgagga aacgatgctg gtcaagctgg 1561 aggcgagcag ttgattgccc atccttgcaa ctcacagctg cttcacgttc tctgcgcctc 1621 gttccatgca ctgcgtcgca cttgtcggcg ggcgaagaca cgaggagagg acaggctgtg 1681 tcctcgagcg agagtagatg ggaaggttac tggaagtcgt gctgttttct atcggcttgc 1741 acgcatctgc tctggttcac tcgctgtgag gcccagcttc catgctgctg cgatctggga 1801 gtgcaagccc gcggtcagca gccattgccc gcctacaatg tcttcccaaa gccgcccttg 1861 ctcgctcgct gcgtggagtt cgacgttctc ctcgctccgc aagcattgga atgaaccttg 1921 ctctctagtt ccctcctccg tgacctcgtt tcgtccttta gacggcacga tggaaggaag 1981 aaatctctgc ggacaagcaa atctgctggc tcgccttgta ggtcgcctac cggagcaagc 2041 cttgtgccgc cgggatgcca acgtcgtttt ttgacgtttg caagacgtag aggacgcttc 2101 ggacgacgaa acaagctgtg aggacatgga agtcgtggga ggaacggcgc agagcggcgc 2161 cgcgggagca taaggcaagc gagatagtcc agaaatcgcg gcgccaagta cagtaattta 2221 ttggagcagg caccagaagc gggcagcagt atgcgcaggc ttggggtcga cgagagacga 2281 ctccctcata ctcggttacc tcgagcaata caatcaatcg aagctgcgcg aatctcggct 2341 tgtaagggtc ggaaaggaac ctcggagatg gccacgtcac atcaccaact tatcgatctc 2401 agccgacgtc gcagagaggg cgagcgaagc ggtgaaggag ggaaacaatc cctcgagagc 2461 atgatccgtc tgaatctgca gcgcaggaag ccgtcacacg cccgcctcga gcgcaggtcg 2521 ggtccagccg ggggacgaaa cgcgcgaggg ctgatttcgt gagcgaagga agccgcatcg 2581 acaagttcgc gtccctttgc cctctttccc atcacccgct ctcgctctac ccgctcagaa 2641 caacaccaga tcagtcacaa tgggcaagga gaagacgcac gtctcgcgcc cgcgcctcaa 2701 ctcgaacatg gacgccgacc tctacggcta caagtggtcc cgcgacaacg tcggccagtc 2761 gggcgccacg atctaccgcc tctacggcaa gccggacgcc ccggagctct tcctcaagca 2821 cggcaagggc tcggtcgcca acgacgtcac ggacgagatg gtccgcctca actggctcac 2881 ggagttcatg ccgctcccga cgatcaagca cttcatccgc acgccggacg acgcctggct 2941 cctcacgacg gccatcccgg gcaagacggc cttccaggtc ctcgaggagt acccggactc 3001 gggcgagaac atcgtcgacg ccctcgccgt cttcctccgc cgcctccact cgatcccggt 3061 ctgcaactgc ccgttcaact cggaccgcgt cttccgcctc gcccaggccc agtcgcgcat 3121 gaacaacggc ctcgtcgacg cctcggactt cgacgacgag cgcaacggct ggccggtcga 3181 gcaggtctgg aaggagatgc acaagctcct cccgttctcg ccggactcgg tcgtcacgca 3241 cggcgacttc tcgctcgaca acctcatctt cgacgagggc aagctcatcg gctgcatcga 3301 cgtcggccgc gtcggcatcg ccgaccgcta ccaggacctc gccatcctct ggaactgcct 3361 cggcgagttc tcgccgtcgc tccagaagcg cctcttccag aagtacggca tcgacaaccc 3421 ggacatgaac aagctccagt tccacctcat gctcgacgag ttcttctaga tcattccact 3481 caacattcag gctcctctgc gcacgtaaag tgccaaaggc aataccctgc tcggtggaat 3541 gccgccgggc ttgtcgattt tacgcacata tgcgcattct tgacttgaag cggaggagtt 3601 cttcgttgcg ggttacagtg ttttaataaa agaatggtca aatcaaactg ctagatatac 3661 ctgtcagaca ctctagttgt tgacccctat actcttaata catcagacag tacatgcatg 3721 ttgcatgatg atgataatgt ctgtttagat tccaagtgtc tactgctggc gtcgacctag 3781 ggggatcctc tagagtcgac ctgcaggcat gcaagcttta agaggagtcc accatggtag 3841 atctactgga ttttggtttt aggaattaga aattttattg atagaagtat tttacaaata 3901 caaatacata ctaagggttt cttatatgct caacacatga gcgaaaccct ataggaaccc 3961 taattccctt atctgggaac tactcacaca ttattatgga gaaaactagt ctactcgtac 4021 ttgacgcgct tgcgggtgcc cgcgtcgccc gcctcgcact cgccgaggcg gcgcttgagt 4081 tcgcggatct cgtcgccgcc gagctgcgag aggtcgatgc gcgtctcgta gaggccggtg 4141 atcgactggt ggatgagggt cgcgtcgagg acttccttgg tcgaggtgta gcgcttgcgg 4201 tcgatggtgg tgtcgaagta cttgaacgcg gcaggggcgc cgaggttggt gagggtgaag 4261 aggtggatga tgttctccgc ctgctcgcgg atgggcttgt cgcggtgctt gttgtaggcc 4321 gagaggacct tgtcgaggtt cgcgtcggcg aggatgacgc gcttcgagaa ctcgctgatc 4381 tgctcgatga tctcgtcgag gtagtgcttg tgctgctcga cgaagagctg cttctgctcg 4441 ttgtcctcgg gcgagccctt gagcttctcg tagtgcgacg cgaggtagag gaagttgacg 4501 tacttcgacg ggagggcgag ttcgttgccc ttctggagtt cgccggccga ggcgagcatg 4561 cgcttgcggc cgttctccaa ctcgaagagc gagtacttgg ggagcttgat gatgaggtcc 4621 ttcttgactt ccttgtagcc cttcgcttcg aggaagtcga tcgggttctt ctcgaacgac 4681 gagcgctcca tgatggtgat gccgaggagt tccttgaccg acttgagctt cttcgacttg 4741 cccttctcga ccttcgcgac gacgaggacc gagtaggcga ccgtcggcga gtcgaatccg 4801 ccgtacttct tcgggtccca gtccttcttg cgggcgatga gcttgtccga gttgcgcttg 4861 gggaggatcg actccttcga gaagccgccg gtctggacct cggtcttctt gacgatgttg 4921 acctgcggca tcgagaggac cttgcggacg gtggcgaagt cgcggccctt gtcccagacg 4981 atctcgcccg tctcgccgtt ggtctcgatg agggggcgct tgcggatctc gccgttggcg 5041 agggtgatct cggtcttgaa gaagttcatg atgttcgagt agaagaagta cttcgcggtg 5101 gccttgccga tctcctgctc cgacttggcg atcatcttgc ggacgtcgta gaccttgtag 5161 tcgccgtaga cgaactccga ttcgagcttc gggtacttct tgatgagcgc ggtgccgacg 5221 accgcgttga ggtaggcgtc gtgggcgtgg tggtagttgt tgatctcgcg gaccttgtag 5281 aactggaagt ccttgcggaa gtccgagacg agcttcgact tgagggtgat gaccttgacc 5341 tcgcggatga gcttgtcgtt ctcgtcgtac ttggtgttca tgcgcgagtc gaggatctgc 5401 gcgacgtgct tggtgatctg gcgggtctcg acgagctggc gcttgatgaa gcccgccttg 5461 tcgagttccg agagtccgcc gcgctccgcc ttcgtgaggt tgtcgaactt gcgctgggtg 5521 atgagcttgg cgttgaggag ctggcgccag tagttcttca tcttcttgac gacttcctcc 5581 gaggggacgt tgtccgactt gccgcggttc ttgtccgagc gggtgaggac cttgttgtcg 5641 atcgagtcgt ccttgaggaa cgactgcggg acgatgtggt ccacgtcgta gtccgagagg 5701 cggttgatgt cgagttcctg gtccacgtac atgtcgcggc cgttctggag gtagtagagg 5761 tagagcttct cgttctggag ctgggtgttc tcgacggggt gctccttgag gatctgcgag 5821 ccgagttcct tgatgccttc ctcgatgcgc ttcatgcgct cgcgcgagtt cttctggccc 5881 ttctgggtgg tctggttctc gcgggccatc tcgatgacga tgttctcggg cttgtggcgg 5941 cccatgacct tgacgagttc gtccacgacc ttgacggtct ggaggatgcc cttcttgatc 6001 gcgggcgagc cggcgaggtt ggcgatgtgc tcgtggagcg agtcgccctg gcccgagacc 6061 tgggccttct ggatgtcttc cttgaaggtg agcgagtcgt cgtggatgag ctgcatgaag 6121 ttgcggttgg cgaagccgtc cgacttgagg aagtcgagga tggtcttgcc cgactgcttg 6181 tcgcggatgc cgttgatgag cttgcgcgag aggcggcccc agcccgtgta tcggcggcgc 6241 ttgagctgct tcatgacctt gtcgtcgaag aggtgggcgt aggtcttgag gcgctcctcg 6301 atcatctcgc ggtcctcgaa gagggtgagg gtgaggacga tgtcttcgag gatgtcctcg 6361 ttctcctcgt tgtcgaggaa gtccttgtcc ttgatgatct tgaggaggtc gtggtaggtg 6421 ccgagcgacg cgttgaagcg gtcctcgacg ccgctgatct cgaccgagtc gaagcactcg 6481 atcttcttga agtagtcttc cttgagctgc ttgacggtga ccttgcggtt ggtcttgaag 6541 aggaggtcca cgatcgcctt cttctgctcg cccgagagga aggccggctt gcgcatgccc 6601 tcggtgacgt acttgacctt ggtgagttcg ttgtagacgg tgaagtactc gtagaggagc 6661 gagtgcttgg ggaggacctt ctcgttcggg aggttcttgt cgaagttggt catgcgctcg 6721 atgaacgact gggccgaggc gcccttgtcc acgacttcct cgaagttcca gggggtgatg 6781 gtctcctccg acttgcgggt catccacgcg aagcgcgagt tgccgcgggc gagggggccg 6841 acgtagtacg ggatgcggaa ggtgaggatc ttctcgatct tctcgcggtt gtccttgagg 6901 aacgggtaga agtcttcctg gcggcggagg atggcgtgga gttcgccgag gtggatctgg 6961 tgggtgatcg agccgttgtc gaaggtgcgc tgcttgcgga ggaggtcctc gcggttgagc 7021 ttgacgagga gttcctcggt gccgtccatc ttttcgagga tcggcttgat gaacttgtag 7081 aactcttcct gcgacgcgcc gccgtcgatg tagcccgcgt agccgttctt cgactggtcg 7141 aagaagattt ccttgtactt ctcggggagc tgctggcgga cgagcgcctt gaggagcgtg 7201 aggtcctggt ggtgctcgtc gtagcgcttg atcatcgacg ccgagagcgg ggccttggtg 7261 atctcggtgt tgacgcggag gatgtccgag aggaggatgg cgtccgagag gttcttcgcg 7321 gcgaggaaga ggtccgcgta ctggtcgccg atctgggcga ggaggttgtc gaggtcgtcg 7381 tcgtaggtgt ccttcgagag ctggagcttc gcgtcctcgg cgaggtcgaa gttcgacttg 7441 aagttggggg tgaggccgag cgagagggcg atgaggttgc cgaagaggcc gttcttcttc 7501 tcgcccggga gctgggcgat gaggttttcg aggcggcgcg acttcgagag gcgggccgag 7561 aggatggcct tggcgtccac gcccgaggcg ttgatggggt tctcctcgaa gagctggttg 7621 taggtctgga cgagctggat gaagagcttg tccacgtccg agttgtccgg gttgaggtcg 7681 ccctcgatga ggaagtggcc gcggaacttg atcatgtgcg cgagggcgag gtagatgagg 7741 cggaggtccg ccttgtaggt cgagtccacg agcttcttgc ggaggtggta gatggtgggg 7801 tacttctcgt ggtaggcgac ctcgtccacg atgttgccga agatcgggtg gcgctcgtgc 7861 ttcttgtctt cctcgacgag gaacgactct tcgaggcggt ggaagaacga gtcgtccacc 7921 ttcgccatct cgttcgagaa gatttcctgg aggtagcaga tgcggttctt gcggcgggtg 7981 tatcggcggc gggccgtgcg cttgaggcgg gtggcctccg ccgtctcgcc cgagtcgaag 8041 aggagggcgc cgatgaggtt cttcttgatc gagtggcggt cggtgttgcc gaggaccttg 8101 aacttcttcg acgggacctt gtactcgtcg gtgatgacgg cccagccgac cgagttggtg 8161 ccgatgtcga ggccgatcga gtacttcttg tccatgtggg agggtgggag ttggacgggt 8221 gagttgctgt gaacctgcag agtagtaagc gagtgactgt agctgtacca gagcgagtgc 8281 gagtgagaag tgcaagttca acgatgcggc cgcacccgac gagcgagtga cgagcagggt 8341 cgacctcgct cacccgcttg cactcgtctg gcgcattcgc tgacaggcta gacagactca 8401 cacaggcttg ttctcccagg gccaagtcga gcggcgcact ccgagcgggt caggcgacga 8461 ccacctcctc cgcttggact cgccagggaa gagtcgaccg actcgaggga tacaagggag 8521 tgggacatga tacgagcaac agagtgcact gcagggatgg acaacacagg cgcagggatc 8581 cgcccgccgt cgctcaccca gcccctcgtc tcgctcgcca gcatctcgac ggagcacttt 8641 cggccctccc ctctcacgct cgactggctg tactcgcttc cattgccgcc aggatgtccc 8701 gtccaacctt tcccacagac gagcgaggac tgggccctgg tcgagcagta ccgcctgctc 8761 tgccaggggt tgacggtcat ggcggagtac gagcgggagc gggcgaggcg ccagcagcgc 8821 ggggaggatc cggatgcgca gccgtgggat cctaggagtg cgtcgcagcg ttctgcctct 8881 tctgccgcgc ttgcttacag ccacttgcct cgacagcctc ggacgacccg gccgtcgtct 8941 tcgccgccga atgcctcttc aacgggccag actgggtcga gtcggaggac cacgcgtcgt 9001 cgccgtccga gcacgacgac gccgcctacg aggtgatcga ctcgccctcg caagcggccg 9061 acgcagcctt cctgggcgtc ctctccgccg tccagcaaca acaacagcgg aacgcctctt 9121 cctcgtcctc gacatccttc acgacctcga tccccctctc cctctcggac ttgacagccc 9181 tctcctccgc cctccccctc tcgtcctcct cctctcaacc gcctcaaacc cacctctcac 9241 cctcccaaat ccccgcagac ggcatcctct ccgactcggc tctctcaccc gcctccctcc 9301 caagcttgat gagcgccatc gccgagttgg agcagtgcgt cgcgcgattg agtttggagg 9361 cgagcgaggc gaggaggttg cagaggcgga tgaggggggc cgtggtcggt gcttcggcgg 9421 cgaatgtggg ggcggtgggt gatggagaag cagaggcggc gaagaagaag ggcaagaaca 9481 ggaagaagaa gaaaaaggtc gccccagccc ctcctcctcc tccccctctc cctccggctg 9541 tccctgcgcc acccgcgccg aaacagcgcg aaggagcagg aggagacgac tcggatttgt 9601 tgaacgcgct gcactcggtt tcgatgctgg atcgcagggc ggacgagtac cgcgagcgat 9661 tgagggttct caaggtccgt ctggcttaag attgaatcct gttgccggtc ttgcgatgat 9721 tatcatataa tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac 9781 gttatttatg agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat 9841 agaaaacaaa atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt 9901 actagatcgg gaattaaact atcagtgttt gacaggatat attggcgggt aaacctaaga 9961 gaaaagagcg tttattagaa taatcggata tttaaaaggg cgtgaaaagg tttatccgtt 10021 cgtccatttg tatgtgcatg ccaaccacag ggttcccctc gggatcaaag tactttgatc 10081 caacccctcc gctgctatag tgcagtcggc ttctgacgtt cagtgcagcc gtcttctgaa 10141 aacgacatgt cgcacaagtc ctaagttacg cgacaggctg ccgccctgcc cttttcctgg 10201 cgttttcttg tcgcgtgttt tagtcgcata aagtagaata cttgcgacta gaaccggaga 10261 cattacgcca tgaacaagag cgccgccgct ggcctgctgg gctatgcccg cgtcagcacc 10321 gacgaccagg acttgaccaa ccaacgggcc gaactgcacg cggccggctg caccaagctg 10381 ttttccgaga agatcaccgg caccaggcgc gaccgcccgg agctggccag gatgcttgac 10441 cacctacgcc ctggcgacgt tgtgacagtg accaggctag accgcctggc ccgcagcacc 10501 cgcgacctac tggacattgc cgagcgcatc caggaggccg gcgcgggcct gcgtagcctg 10561 gcagagccgt gggccgacac caccacgccg gccggccgca tggtgttgac cgtgttcgcc 10621 ggcattgccg agttcgagcg ttccctaatc atcgaccgca cccggagcgg gcgcgaggcc 10681 gccaaggccc gaggcgtgaa gtttggcccc cgccctaccc tcaccccggc acagatcgcg 10741 cacgcccgcg agctgatcga ccaggaaggc cgcaccgtga aagaggcggc tgcactgctt 10801 ggcgtgcatc gctcgaccct gtaccgcgca cttgagcgca gcgaggaagt gacgcccacc 10861 gaggccaggc ggcgcggtgc cttccgtgag gacgcattga ccgaggccga cgccctggcg 10921 gccgccgaga atgaacgcca agaggaacaa gcatgaaacc gcaccaggac ggccaggacg 10981 aaccgttttt cattaccgaa gagatcgagg cggagatgat cgcggccggg tacgtgttcg 11041 agccgcccgc gcacgtctca accgtgcggc tgcatgaaat cctggccggt ttgtctgatg 11101 ccaagctggc ggcctggccg gccagcttgg ccgctgaaga aaccgagcgc cgccgtctaa 11161 aaaggtgatg tgtatttgag taaaacagct tgcgtcatgc ggtcgctgcg tatatgatgc 11221 gatgagtaaa taaacaaata cgcaagggga acgcatgaag gttatcgctg tacttaacca 11281 gaaaggcggg tcaggcaaga cgaccatcgc aacccatcta gcccgcgccc tgcaactcgc 11341 cggggccgat gttctgttag tcgattccga tccccagggc agtgcccgcg attgggcggc 11401 cgtgcgggaa gatcaaccgc taaccgttgt cggcatcgac cgcccgacga ttgaccgcga 11461 cgtgaaggcc atcggccggc gcgacttcgt agtgatcgac ggagcgcccc aggcggcgga 11521 cttggctgtg tccgcgatca aggcagccga cttcgtgctg attccggtgc agccaagccc 11581 ttacgacata tgggccaccg ccgacctggt ggagctggtt aagcagcgca ttgaggtcac 11641 ggatggaagg ctacaagcgg cctttgtcgt gtcgcgggcg atcaaaggca cgcgcatcgg 11701 cggtgaggtt gccgaggcgc tggccgggta cgagctgccc attcttgagt cccgtatcac 11761 gcagcgcgtg agctacccag gcactgccgc cgccggcaca accgttcttg aatcagaacc 11821 cgagggcgac gctgcccgcg aggtccaggc gctggccgct gaaattaaat caaaactcat 11881 ttgagttaat gaggtaaaga gaaaatgagc aaaagcacaa acacgctaag tgccggccgt 11941 ccgagcgcac gcagcagcaa ggctgcaacg ttggccagcc tggcagacac gccagccatg 12001 aagcgggtca actttcagtt gccggcggag gatcacacca agctgaagat gtacgcggta 12061 cgccaaggca agaccattac cgagctgcta tctgaataca tcgcgcagct accagagtaa 12121 atgagcaaat gaataaatga gtagatgaat tttagcggct aaaggaggcg gcatggaaaa 12181 tcaagaacaa ccaggcaccg acgccgtgga atgccccatg tgtggaggaa cgggcggttg 12241 gccaggcgta agcggctggg ttgcctgccg gccctgcaat ggcactggaa cccccaagcc 12301 cgaggaatcg gcgtgagcgg tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg 12361 ggtgatgacc tggtggagaa gttgaaggcc gcgcaggccg cccagcggca acgcatcgag 12421 gcagaagcac gccccggtga atcgtggcaa gcggccgctg atcgaatccg caaagaatcc 12481 cggcaaccgc cggcagccgg tgcgccgtcg attaggaagc cgcccaaggg cgacgagcaa 12541 ccagattttt tcgttccgat gctctatgac gtgggcaccc gcgatagtcg cagcatcatg 12601 gacgtggccg ttttccgtct gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac 12661 gagcttccag acgggcacgt agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg 12721 gattacgacc tggtactgat ggcggtttcc catctaaccg aatccatgaa ccgataccgg 12781 gaagggaagg gagacaagcc cggccgcgtg ttccgtccac acgttgcgga cgtactcaag 12841 ttctgccggc gagccgatgg cggaaagcag aaagacgacc tggtagaaac ctgcattcgg 12901 ttaaacacca cgcacgttgc catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg 12961 acggtatccg agggtgaagc cttgattagc cgctacaaga tcgtaaagag cgaaaccggg 13021 cggccggagt acatcgagat cgagctagct gattggatgt accgcgagat cacagaaggc 13081 aagaacccgg acgtgctgac ggttcacccc gattactttt tgatcgatcc cggcatcggc 13141 cgttttctct accgcctggc acgccgcgcc gcaggcaagg cagaagccag atggttgttc 13201 aagacgatct acgaacgcag tggcagcgcc ggagagttca agaagttctg tttcaccgtg 13261 cgcaagctga tcgggtcaaa tgacctgccg gagtacgatt tgaaggagga ggcggggcag 13321 gctggcccga tcctagtcat gcgctaccgc aacctgatcg agggcgaagc atccgccggt 13381 tcctaatgta cggagcagat gctagggcaa attgccctag caggggaaaa aggtcgaaaa 13441 ggactctttc ctgtggatag cacgtacatt gggaacccaa agccgtacat tgggaaccgg 13501 aacccgtaca ttgggaaccc aaagccgtac attgggaacc ggtcacacat gtaagtgact 13561 gatataaaag agaaaaaagg cgatttttcc gcctaaaact ctttaaaact tattaaaact 13621 cttaaaaccc gcctggcctg tgcataactg tctggccagc gcacagccga agagctgcaa 13681 aaagcgccta cccttcggtc gctgcgctcc ctacgccccg ccgcttcgcg tcggcctatc 13741 gcggccgctg gccgctcaaa aatggctggc ctacggccag gcaatctacc agggcgcgga 13801 caagccgcgc cgtcgccact cgaccgccgg cgcccacatc aaggcaccct gcctcgcgcg 13861 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg 13921 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg 13981 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac 14041 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac 14101 agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg 14161 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 14221 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 14281 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 14341 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 14401 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 14461 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 14521 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 14581 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 14641 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 14701 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca 14761 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 14821 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 14881 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 14941 cagtggaacg aaaactcacg ttaagggctg atgaatcccc taatgatttt tatcaaaatc 15001 attaagttaa ggtagataca catcttgtca tatgatcaaa tggtttcgcc aaaaatcaat 15061 aatcagacaa caaaatgtgc gaactcgata ttttacacga ctctctttac caattctgcc 15121 ccgaattaca cttaaaacga ctcaacagct taacgttggc ttgccacgcc ttacttgact 15181 gtaaaactct cactcttacc gaacttggcc gtaacctgcc aaccaaagcg agaacaaaac 15241 ataacatcaa acgaatcgac cgattgttag gtaatcgtca cctccacaaa gagcgactcg 15301 ctgtataccg ttggcatgct agctttatct gttcgggcaa tacgatgccc attgtacttg 15361 ttgactggtc tgatatccgt gagcaaaaac ggcttatggt attgcgagct tcagtcgcac 15421 tacacggtcg ttctgttact ctttatgaga aagcgttccc gctttcagag caatgttcaa 15481 agaaagctca tgaccaattt ctagccgacc ttgcgagcat tctaccgagt aacaccacac 15541 cgctcattgt cagtgatgct ggctttaaag tgccatggta taaatccgtt gagaagctgg 15601 gttggtactg gttaagtcga gtaagaggaa aagtacaata tgcagaccta ggagcggaaa 15661 actggaaacc tatcagcaac ttacatgata tgtcatctag tcactcaaag actttaggct 15721 ataagaggct gactaaaagc aatccaatct catgccaaat tctattgtat aaatctcgct 15781 ctaaaggccg aaaaaatcag cgctcgacac ggactcattg tcaccacccg tcacctaaaa 15841 tctactcagc gtcggcaaag gagccatgga ttctagcaac taacttacct gttgaaattc 15901 gaacacccaa acaacttgtt aatatctatt cgaagcgaat gcagattgaa gaaaccttcc 15961 gagacttgaa aagtcctgcc tacggactag gcctacgcca tagccgaacg agcagctcag 16021 agcgttttga tatcatgctg ctaatcgccc tgatgcttca actaacatgt tggcttgcgg 16081 gcgttcatgc tcagaaacaa ggttgggaca agcacttcca ggctaacaca gtcagaaatc 16141 gaaacgtact ctcaacagtt cgcttaggca tggaagtttt gcggcattct ggctacacaa 16201 taacaaggga agacttactc gtggctgcaa ccctactagc tcaaaattta ttcacacatg 16261 gttacgcttt ggggaaatta tgaggggatc tctcagcgtt aagggatttt ggtcatgcat 16321 tctaggtact aaaacaattc atccagtaaa atataatatt ttattttctc ccaatcaggc 16381 ttgatcccca gtaagtcaaa aaatagctcg acatactgtt cttccccgat atcctccctg 16441 atcgaccgga cgcagaaggc aatgtcatac cacttgtccg ccctgccgct tctcccaaga 16501 tcaataaagc cacttacttt gccatctttc acaaagatgt tgctgtctcc caggtcgccg 16561 tgggaaaaga caagttcctc ttcgggcttt tccgtcttta aaaaatcata cagctcgcgc 16621 ggatctttaa atggagtgtc ttcttcccag ttttcgcaat ccacatcggc cagatcgtta 16681 ttcagtaagt aatccaattc ggctaagcgg ctgtctaagc tattcgtata gggacaatcc 16741 gatatgtcga tggagtgaaa gagcctgatg cactccgcat acagctcgat aatcttttca 16801 gggctttgtt catcttcata ctcttccgag caaaggacgc catcggcctc actcatgagc 16861 agattgctcc agccatcatg ccgttcaaag tgcaggacct ttggaacagg cagctttcct 16921 tccagccata gcatcatgtc cttttcccgt tccacatcat aggtggtccc tttataccgg 16981 ctgtccgtca tttttaaata taggttttca ttttctccca ccagcttata taccttagca 17041 ggagacattc cttccgtatc ttttacgcag cggtattttt cgatcagttt tttcaattcc 17101 ggtgatattc tcattttagc catttattat ttccttcctc ttttctacag tatttaaaga 17161 taccccaaga agctaattat aacaagacga actccaattc actgttcctt gcattctaaa 17221 accttaaata ccagaaaaca gctttttcaa agttgttttc aaagttggcg tataacatag 17281 tatcgacgga gccgattttg aaaccgcggt tagttttgct ggccgcatct tctcaaatat 17341 gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct tccctttgca 17401 aatagtcctc ttccaacaat aataatgtca gatcctgtag acaccacatc atccacggtt 17461 ctatactgtt gacccaatgc gtctcccttg tcatctaaac ccacaccggg tgtcataatc 17521 aaccaatcgt aaccttcatc tcctccaccc atgtctcttt gagcaataaa gccgataaca 17581 aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg 17641 gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct 17701 tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg tgcattcgta 17761 atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta 17821 ccaatgtcag caaattttct gtcttcgaag agtaaaaaat tgtacttggc ggataatgcc 17881 tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac atgtgttttt 17941 agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga 18001 acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca 18061 gcaacaggac taggatgagt agcagcacgt tccttatatg tagctttcga catgatttat 18121 cttcgtttcc tgcaggtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat 18181 gtttcttcaa cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt 18241 tcttccttct gttcggagat taccgaatca aaaaaatttc aaggaaaccg aaatcaaaaa 18301 aaagaataaa aaaaaaatga tgaattgaaa agctagctta tcgatgataa gctgtcaaac 18361 atgagaatta attccacgga ctatagacta tacctagtat actccgtcta ctgtacgata 18421 cacttccgct caggtccttg tcctttaacg aggccttacc actcttttgt tactctattg 18481 atccagctca gcaaaggcag tgtgatctaa gattctatct tcgcgatgta gtaaaactag 18541 ctagaccgag aaagagacta gaaatgcaaa aggcacttct acaatggctg ccatcattat 18601 tatccgatgt gacgctgcag cttctcaatg atattcgaat acgctttgag gagatacagc 18661 ctaatatccg acaaactgtt ttacagattt acgatcgtac ttgttaccca tcattgaatt 18721 ttgaacatcc gaacctggga gttttccctg aaacagatag tatatttgaa cctgtataat 18781 aatatatagt ctagcgcttt acggaagaca atgtatgtat ttcggttcct ggagaaacta 18841 ttgcatctat tgcataggta atcttgcacg tcgcatcccc ggttcatttt ctgcgtttcc 18901 atcttgcact tcaatagcat atctttgtta acgaagcatc tgtgcttcat tttgtagaac 18961 aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag 19021 aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt 19081 aaaacaaaaa tgcaacgcga ga //