LOCUS Exported 11257 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Level2 MoClo construct, containing level1 plant expression cassettes for Sp-eCas9 1.1(pEPOR1CB0011) and sgRNA_NbPDS+1 mismatch(pEPOR1CB021). ACCESSION . VERSION . KEYWORDS pEPOR2KN0031 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11257) AUTHORS Raitskin O, Schudoma C, West A, Patron NJ TITLE Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. JOURNAL PLoS One. 2019 Feb 27;14(2):e0211598. doi: 10.1371/journal.pone.0211598. eCollection 2019. PUBMED 30811422 REFERENCE 2 (bases 1 to 11257) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11257 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 454..799 /label=CaMV 35S promoter /note="strong constitutive promoter from cauliflower mosaic virus" primer_bind 742..762 /label=35S promoter /note="CaMV 35S promoter, forward primer" CDS 890..4993 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 5006..5026 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 5033..5752 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(5078..5099) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(5339..5358) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5686..5707 /label=EGFP-C /note="EGFP, forward primer" terminator 5765..6017 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" misc_RNA 6269..6344 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" protein_bind complement(6356..6380) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" misc_feature 6449..6473 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS 6782..7411 /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" CDS 7840..8913 /codon_start=1 /product="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 RepA /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW QAAADRIRKESRQPPAAGASSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI LVMRYRNLIEGEASAGS" rep_origin 8979..9173 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" promoter 9381..9485 /gene="bla" /label=AmpR promoter CDS 9486..10280 /codon_start=1 /gene="aphA-3" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF" rep_origin 10451..11039 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 10940..10959 /label=pBR322ori-F /note="pBR322 origin, forward primer" misc_feature 11175..11199 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" ORIGIN 1 gaacactctg tgccgatgta tcacctctcg caagaattca agcttggagg tcaacatggt 61 ggagcacgac actctggtct actccaaaaa tgtcaaagat acagtctcag aagatcaaag 121 ggctattgag acttttcaac aaaggataat ttcgggaaac ctcctcggat tccattgccc 181 agctatctgt cacttcatcg aaaggacagt agaaaaggaa ggtggctcct acaaatgcca 241 tcattgcgat aaaggaaagg ctatcattca agatctctct gccgacagtg gtcccaaaga 301 tggaccccca cccacgagga gcatcgtgga aaaagaagag gttccaacca cgtctacaaa 361 gcaagtggat tgatgtgata acatggtgga gcacgacact ctggtctact ccaaaaatgt 421 caaagataca gtctcagaag atcaaagggc tattgagact tttcaacaaa ggataatttc 481 gggaaacctc ctcggattcc attgcccagc tatctgtcac ttcatcgaaa ggacagtaga 541 aaaggaaggt ggctcctaca aatgccatca ttgcgataaa ggaaaggcta tcattcaaga 601 tctctctgcc gacagtggtc ccaaagatgg acccccaccc acgaggagca tcgtggaaaa 661 agaagaggtt ccaaccacgt ctacaaagca agtggattga tgtgacatct ccactgacgt 721 aagggatgac gcacaatccc actatccttc gcaagaccct tcctctatat aaggaagttc 781 atttcatttg gagaggacac gctcgagtat aagagctcat ttttacaaca attaccaaca 841 acaacaaaca acaaacaaca ttacaattac atttacaatt atcgatacaa tggacaagaa 901 gtactccatt gggctcgata tcggcacaaa cagcgtcggc tgggccgtca ttacggacga 961 gtacaaggtg ccgagcaaaa aattcaaagt tctgggcaat accgatcgcc acagcataaa 1021 gaagaacctc attggcgccc tcctgttcga ctccggggag acggccgaag ccacgcggct 1081 caaaagaaca gcacggcgca gatatacccg cagaaagaat cggatctgct acctgcagga 1141 gatctttagt aatgagatgg ctaaggtgga tgactctttc ttccataggc tggaggagtc 1201 ctttttggtg gaggaggata aaaagcacga gcgccaccca atctttggca atatcgtgga 1261 cgaggtggcg taccatgaaa agtacccaac catatatcat ctgaggaaga agcttgtaga 1321 cagtactgat aaggctgact tgcggttgat ctatctcgcg ctggcgcata tgatcaaatt 1381 tcggggacac ttcctcatcg agggggacct gaacccagac aacagcgatg tcgacaaact 1441 ctttatccaa ctggttcaga cttacaatca gcttttcgaa gagaacccga tcaacgcatc 1501 cggagttgac gccaaagcaa tcctgagcgc taggctgtcc aaatcccggc ggctcgaaaa 1561 cctcatcgca cagctccctg gggagaagaa gaacggcctg tttggtaatc ttatcgccct 1621 gtcactcggg ctgaccccca actttaaatc taacttcgac ctggccgaag atgccaagct 1681 tcaactgagc aaagacacct acgatgatga tctcgacaat ctgctggccc agatcggcga 1741 ccagtacgca gacctttttt tggcggcaaa gaacctgtca gacgccattc tgctgagtga 1801 tattctgcga gtgaacacgg agatcaccaa agctccgctg agcgctagta tgatcaagcg 1861 ctatgatgag caccaccaag acttgacttt gctgaaggcc cttgtcagac agcaactgcc 1921 tgagaagtac aaggaaattt tcttcgatca gtctaaaaat ggctacgccg gatacattga 1981 cggcggagca agccaggagg aattttacaa atttattaag cccatcttgg aaaaaatgga 2041 cggcaccgag gagctgctgg taaagcttaa cagagaagat ctgttgcgca aacagcgcac 2101 tttcgacaat ggaagcatcc cccaccagat tcacctgggc gaactgcacg ctatcctcag 2161 gcggcaagag gatttctacc cctttttgaa agataacagg gaaaagattg agaaaatcct 2221 cacatttcgg ataccctact atgtaggccc cctcgcccgg ggaaattcca gattcgcgtg 2281 gatgactcgc aaatcagaag agactatcac tccctggaac ttcgaggaag tcgtggataa 2341 gggggcctct gcccagtcct tcatcgaaag gatgactaac tttgataaaa atctgcctaa 2401 cgaaaaggtg cttcctaaac actctctgct gtacgagtac ttcacagttt ataacgagct 2461 caccaaggtc aaatacgtca cagaagggat gagaaagcca gcattcctgt ctggagagca 2521 gaagaaagct atcgtggacc tcctcttcaa gacgaaccgg aaagttaccg tgaaacagct 2581 caaagaagat tatttcaaaa agattgaatg tttcgactct gttgaaatca gcggagtgga 2641 ggatcgcttc aacgcatccc tgggaacgta tcacgatctc ctgaaaatca ttaaagacaa 2701 ggacttcctg gacaatgagg agaacgagga cattcttgag gacattgtcc tcacccttac 2761 gttgtttgaa gatagggaga tgattgaaga acgcttgaaa acttacgctc atctcttcga 2821 cgacaaagtc atgaaacagc tcaagaggcg ccgatataca ggatgggggc ggctgtcaag 2881 aaaactgatc aatgggatcc gagacaagca gagtggaaag acaatcctgg attttcttaa 2941 gtccgatgga tttgccaacc ggaacttcat gcagttgatc catgatgact ctctcacctt 3001 taaggaggac atccagaaag cacaagtttc tggccagggg gacagtctcc acgagcacat 3061 cgctaatctt gcaggtagcc cagctatcaa aaagggaata ctgcagaccg ttaaggtcgt 3121 ggatgaactc gtcaaagtaa tgggaaggca taagcccgag aatatcgtta tcgagatggc 3181 ccgagagaac caaactaccc agaagggaca gaagaacagt agggaaagga tgaagaggat 3241 tgaagagggt ataaaagaac tggggtccca aatccttaag gaacacccag ttgaaaacac 3301 ccagcttcag aatgagaagc tctacctgta ctacctgcag aacggcaggg acatgtacgt 3361 ggatcaggaa ctggacatca atcggctctc cgactacgac gtggatcata tcgtgcccca 3421 gtcttttctc gcagatgatt ctattgataa taaagtgttg acaagatccg ataaaaatag 3481 agggaagagt gataacgtcc cctcagaaga agttgtcaag aaaatgaaaa attattggcg 3541 gcagctgctg aacgccaaac tgatcacaca acggaagttc gataatctga ctaaggctga 3601 acgaggtggc ctgtctgagt tggataaagc cggcttcatc aaaaggcagc ttgttgagac 3661 acgccagatc accaagcacg tggcccaaat tctcgattca cgcatgaaca ccaagtacga 3721 tgaaaatgac aaactgattc gagaggtgaa agttattact ctgaagtcta agctggtttc 3781 agatttcaga aaggactttc agttttataa ggtgagagag atcaacaatt accaccatgc 3841 gcatgatgcc tacctgaatg cagtggtagg cactgcactt atcaaaaaat atcccgcgct 3901 tgaatctgaa tttgtttacg gagactataa agtgtacgat gttaggaaaa tgatcgcaaa 3961 gtctgagcag gaaataggca aggccaccgc taagtacttc ttttacagca atattatgaa 4021 ttttttcaag accgagatta cactggccaa tggagagatt cggaaggcac cacttatcga 4081 aacaaacgga gaaacaggag aaatcgtgtg ggacaagggt agggatttcg cgacagtccg 4141 gaaggtcctg tccatgccgc aggtgaacat cgttaaaaag accgaagtac agaccggagg 4201 cttctccaag gaaagtatcc tcccgaaaag gaacagcgac aagctgatcg cacgcaaaaa 4261 agattgggac cccaagaaat acggcggatt cgattctcct acagtcgctt acagtgtact 4321 ggttgtggcc aaagtggaga aagggaagtc taaaaaactc aaaagcgtca aggaactgct 4381 gggcatcaca atcatggagc gatcaagctt cgaaaaaaac cccatcgact ttctcgaggc 4441 gaaaggatat aaagaggtca aaaaagacct catcattaag cttcccaagt actctctctt 4501 tgagcttgaa aacggccgga aacgaatgct cgctagtgcg ggcgagctgc agaaaggtaa 4561 cgagctggca ctgccctcta aatacgttaa tttcttgtat ctggccagcc actatgaaaa 4621 gctcaaagga tctcccgaag ataatgagca gaagcagctg ttcgtggaac aacacaaaca 4681 ctaccttgat gagatcatcg agcaaataag cgaattctcc aaaagagtga tcctcgccga 4741 cgctaacctc gataaggtgc tttctgctta caataagcac agggataagc ccatcaggga 4801 gcaggcagaa aacattatcc acttgtttac tctgaccaac ttgggcgcgc ctgcagcctt 4861 caagtacttc gacaccacca tagacagaaa gcggtacacc tctacaaagg aggtcctgga 4921 cgccacactg attcatcagt caattacggg gctctatgaa acaagaatcg acctctctca 4981 gctcggtgga gacagcaggg ctgaccccaa gaagaagagg aaggtgggtt cgatggtgag 5041 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt 5101 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct 5161 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac 5221 caccttcggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga agcagcacga 5281 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga 5341 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg 5401 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga 5461 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa 5521 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta 5581 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag 5641 ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga 5701 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt gagcttgtca 5761 agcagatcgt tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct 5821 tgcgatgatt atcatataat ttctgttgaa ttacgttaag catgtaataa ttaacatgta 5881 atgcatgacg ttatttatga gatgggtttt tatgattaga gtcccgcaat tatacattta 5941 atacgcgata gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc 6001 atctatgtta ctagatcgac gctactagaa ttcgagctcg gagcatcttc attcttaaga 6061 tatgaagata atcttcaaaa ggcccctggg aatctgaaag aagagaagca ggcccattta 6121 tatgggaaag aacaatagta tttcttatat aggcccattt aagttgaaaa caatcttcaa 6181 aagtcccaca tcgcttagat aagaaaacga agctgagttt atatacagct agagtcgaag 6241 tagtgattgc cgttaatttg agagttcagt tttagagcta gaaatagcaa gttaaaataa 6301 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tctagaccca 6361 gctttcttgt acaaagttgg cattacgctt tacgaggatg cacatgtgac cgagggacac 6421 gaagtgatcc gtttaaacta tcagtgtttg acaggatata ttggcgggta aacctaagag 6481 aaaagagcgt ttattagaat aatcggatat ttaaaagggc gtgaaaaggt ttatccgttc 6541 gtccatttgt atgtgcatgc caaccacagg gttcccctcg ggagtcagcc gtgcggctgc 6601 atgaaatcct ggccggtttg tctgatgcca agctggcggc ctggccggcc agcttggccg 6661 ctgaagaaac cgagcgccgc cgtctaaaaa ggtgatgtgt atttgagtaa aacagcttgc 6721 gtcatgcggt cgctgcgtat atgatgcgat gagtaaataa acaaatacgc aaggggaacg 6781 catgaaggtt atcgctgtac ttaaccagaa aggcgggtca ggcaagacga ccatcgcaac 6841 ccatctagcc cgcgccctgc aactcgccgg ggccgatgtt ctgttagtcg attccgatcc 6901 ccagggcagt gcccgcgatt gggcggccgt gcgggaagat caaccgctaa ccgttgtcgg 6961 catcgaccgc ccgacgattg accgcgacgt gaaggccatc ggccggcgcg acttcgtagt 7021 gatcgacgga gcgccccagg cggcggactt ggctgtgtcc gcgatcaagg cagccgactt 7081 cgtgctgatt ccggtgcagc caagccctta cgacatatgg gccaccgccg acctggtgga 7141 gctggttaag cagcgcattg aggtcacgga tggaaggcta caagcggcct ttgtcgtgtc 7201 gcgggcgatc aaaggcacgc gcatcggcgg tgaggttgcc gaggcgctgg ccgggtacga 7261 gctgcccatt cttgagtccc gtatcacgca gcgcgtgagc tacccaggca ctgccgccgc 7321 cggcacaacc gttcttgaat cagaacccga gggcgacgct gcccgcgagg tccaggcgct 7381 ggccgctgaa attaaatcaa aactcatttg agttaatgag gtaaagagaa aatgagcaaa 7441 agcacaaaca cgctaagtgc cggccgtccg agcgcacgca gcagcaaggc tgcaacgttg 7501 gccagcctgg cagacacgcc agccatgaag cgggtcaact ttcagttgcc ggcggaggat 7561 cacaccaagc tgaagatgta cgcggtacgc caaggcaaga ccattaccga gctgctatct 7621 gaatacatcg cgcagctacc agagtaaatg agcaaatgaa taaatgagta gatgaatttt 7681 agcggctaaa ggaggcggca tggaaaatca agaacaacca ggcaccgacg ccgtggaatg 7741 ccccatgtgt ggaggaacgg gcggttggcc aggcgtaagc ggctgggttg tctgccggcc 7801 ctgcaatggc actggaaccc ccaagcccga ggaatcggcg tgagcggtcg caaaccatcc 7861 ggcccggtac aaatcggcgc ggcgctgggt gatgacctgg tggagaagtt gaaggccgcg 7921 caggccgccc agcggcaacg catcgaggca gaagcacgcc ccggtgaatc gtggcaagcg 7981 gccgctgatc gaatccgcaa agaatcccgg caaccgccgg cagccggtgc gtcgtcgatt 8041 aggaagccgc ccaagggcga cgagcaacca gattttttcg ttccgatgct ctatgacgtg 8101 ggcacccgcg atagtcgcag catcatggac gtggccgttt tccgtctgtc gaagcgtgac 8161 cgacgagctg gcgaggtgat ccgctacgag cttccagacg ggcacgtaga ggtttccgca 8221 gggccggccg gcatggccag tgtgtgggat tacgacctgg tactgatggc ggtttcccat 8281 ctaaccgaat ccatgaaccg ataccgggaa gggaagggag acaagcccgg ccgcgtgttc 8341 cgtccacacg ttgcggacgt actcaagttc tgccggcgag ccgatggcgg aaagcagaaa 8401 gacgacctgg tagaaacctg cattcggtta aacaccacgc acgttgccat gcagcgtacg 8461 aagaaggcca agaacggccg cctggtgacg gtatccgagg gtgaagcctt gattagccgc 8521 tacaagatcg taaagagcga aaccgggcgg ccggagtaca tcgagatcga gctagctgat 8581 tggatgtacc gcgagattac agaaggcaag aacccggacg tgctgacggt tcaccccgat 8641 tactttttga tcgatcccgg catcggccgt tttctctacc gcctggcacg ccgcgccgca 8701 ggcaaggcag aagccagatg gttgttcaag acgatctacg aacgcagtgg cagcgccgga 8761 gagttcaaga agttctgttt caccgtgcgc aagctgatcg ggtcaaatga cctgccggag 8821 tacgatttga aggaggaggc ggggcaggct ggcccgatcc tagtcatgcg ctaccgcaac 8881 ctgatcgagg gcgaagcatc cgccggttcc taatgtacgg agcagatgct agggcaaatt 8941 gccctagcag gggaaaaagg tcgaaaaagc ttctttcctg tggatagcac gtacattggg 9001 aacccaaagc cgtacattgg gaaccggaac ccgtacattg ggaacccaaa gccgtacatt 9061 gggaaccggt cacacatgta agtgactgat ataaaagaga aaaaaggcga tttttccgcc 9121 taaaactctt taaaacttat taaaactctt aaaacccgcc tggcctgtgc ataactgtct 9181 ggccagcgca cagccgaaca gctgcaaaaa gcgcccacat caaggctccg agtgcgcgga 9241 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 9301 cccgcctggc ctgtgcataa ctgtctggcc agcgcacagc cgaacagctg caaaaagcgc 9361 ccacatcaag gctccgagtg cgcggaaccc ctatttgttt atttttctaa atacattcaa 9421 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga 9481 agagtatggc taaaatgaga atatcaccgg aattgaaaaa actgatcgaa aaataccgct 9541 gcgtaaaaga tacggaagga atgtctcctg ctaaggtata taagctggtg ggagaaaatg 9601 aaaacctata tttaaaaatg acggacagcc ggtataaagg gaccacctat gatgtggaac 9661 gggaaaagga catgatgcta tggctggaag gaaagctgcc tgttccaaag gtcctgcact 9721 ttgaacggca tgatggctgg agcaatctgc tcatgagtga ggccgatggc gtcctttgct 9781 cggaagagta tgaagatgaa caaagccctg aaaagattat cgagctgtat gcggagtgca 9841 tcaggctctt tcactccatc gacatatcgg attgtcccta tacgaatagc ttagacagcc 9901 gcttagccga attggattac ttactgaata acgatctggc cgatgtggat tgcgaaaact 9961 gggaagagga cactccattt aaagatccgc gcgagctgta tgatttttta aagacggaaa 10021 agcccgaaga ggaacttgtc ttttcccacg gcgacctggg agacagcaac atctttgtga 10081 aagatggcaa agtaagtggc tttattgatc ttgggagaag cggcagggcg gacaagtggt 10141 atgacattgc cttctgcgtc cggtcgctca gggaggatat cggggaagaa cagtatgtcg 10201 agctattttt tgacttactg gggatcaagc ctgattggga gaaaataaaa tattatattt 10261 tactggatga attgttttag ctgtcagacc aagtttactc atatatactt tagattgatt 10321 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga 10381 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca 10441 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac 10501 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg 10561 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag 10621 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac 10681 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt 10741 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg 10801 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc 10861 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc 10921 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc 10981 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa 11041 acgccagcaa cgcggccttt ttacggttcc tgctcggatc tgttggaccg gacagtagtc 11101 atggttgatg ggctgcctgt atcgagtggt gattttgtgc cgagctgccg gtcggggagc 11161 tgttggctgg ctggtggcag gatatattgt ggtgtaaaca aattgacgct tagacaactt 11221 aataacacat tgcggacgtt tttaatgtac tggggtt //