LOCUS Exported 16084 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION For CRISPR/Cas9 mediated gene editing in Arabidopsis; provides a visual screen for Cas9-free plants. Enable production of two gRNAs. ACCESSION . VERSION . KEYWORDS pHDE-35S-Cas9-mCherry-UBQ SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 16084) AUTHORS Gao X, Chen J, Dai X, Zhang D, Zhao Y TITLE An effective strategy for reliably isolating heritable and Cas9-free Arabidopsis mutants generated by CRISPR/Cas9-mediated genome editing. JOURNAL Plant Physiol. 2016 May 15. pii: pp.00663.2016. PUBMED 27208253 REFERENCE 2 (bases 1 to 16084) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..16084 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 1..25 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" primer_bind complement(632..650) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 900..919 /label=mCherry-F /note="mCherry, forward primer" terminator 1203..1455 /label=MAS terminator /note="mannopine synthase terminator" promoter 2568..2747 /label=NOS promoter /note="nopaline synthase promoter" promoter 2600..2783 /function="NOS promoter" /label=NOS promoter /note="nopaline synthase promoter" terminator 3898..4150 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" promoter 4283..4628 /label=CaMV 35S promoter /note="strong constitutive promoter from cauliflower mosaic virus" primer_bind 4571..4591 /label=35S promoter /note="CaMV 35S promoter, forward primer" CDS 4633..8736 /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 LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 8749..8769 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(9281..9298) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(9281..9297) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(9290..9312) /label=M13/pUC Forward /note="In lacZ gene" misc_feature 9519..9543 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" CDS 10071..10862 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 11106..11694 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 11595..11614 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 11848..11865 /label=L4440 /note="L4440 vector, forward primer" misc_feature 11880..12020 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(11946..11965) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 12106..12128 /label=pGEX 3' /note="pGEX vectors, reverse primer" rep_origin 12364..12558 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" CDS complement(12624..13691) /codon_start=1 /product="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 RepA /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV MRYRNLIEGEASAGS" CDS complement(14125..14754) /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" ORIGIN 1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactagtgaa gtgaagcttg 61 gtctcaacta tgagacctaa gctggcacaa ctatatttcc aacatcacta gctaccatca 121 aaagattgac ttctcatctt actcgattga aaccaaatta acatagggtt tttatttaaa 181 taaaagttta accttctttt taaaaaattg ttcatagtgt catgtcagaa caagagctac 241 aaatcacaca tagcatgcat aagcggagct atgatgagtg gtattgtttt gttcgtcact 301 tgtcactctt ttccaacaca taatcccgac aacaacgtaa gagcatctct ctctctccac 361 acacactcat gcatgcatgc attcttacac gtgattgcca tgcaaatctc ctttctcacc 421 tataaataca aaccaaccct tcactacact cttcactcaa accaaaacaa gaaaacatac 481 acaaatagca aaacggtacc aacaatggat aacatggcca tcatcaagga gttcatgcgc 541 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc 601 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc 661 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg 721 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg 781 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg 841 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc 901 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag 961 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac 1021 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac 1081 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag 1141 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtgagag 1201 ctcttggact cccatgttgg caaaggcaac caaacaaaca atgaatgatc cgctcctgca 1261 tatggggcgg tttgagtatt tcaactgcca tttgggctga attgaagaca tgctcctgtc 1321 agaaattccg tgatcttact caatattcag taatctcggc caatatccta aatgtgcgtg 1381 gctttatctg tctttgtatt gtttcatcaa ttcatgtaac gtttgctttt cttatgaatt 1441 ttcaaataaa ttatcgtatt gagaccgaat tctcgtcgac gagtcagtaa taaacggcgt 1501 caaagtggtt gcagccggca cacacgagtc gtgtttatca actcaaagca caaatacttt 1561 tcctcaacct aaaaataagg caattagcca aaaacaactt tgcgtgtaaa caacgctcaa 1621 tacacgtgtc attttattat tagctattgc ttcaccgcct tagctttctc gtgacctagt 1681 cgtcctcgtc ttttcttctt cttcttctat aaaacaatac ccaaagagct cttcttcttc 1741 acaattcaga tttcaatttc tcaaaatctt aaaaactttc tctcaattct ctctaccgtg 1801 atcaaggtaa atttctgtgt tccttattct ctcaaaatct tcgattttgt tttcgttcga 1861 tcccaatttc gtatatgttc tttggtttag attctgttaa tcttagatcg aagacgattt 1921 tctgggtttg atcgttagat atcatcttaa ttctcgatta gggtttcata gatatcatcc 1981 gatttgttca aataatttga gttttgtcga ataattactc ttcgatttgt gatttctatc 2041 tagatctggt gttagtttct agtttgtgcg atcgaatttg tcgattaatc tgagtttttc 2101 tgattaacag ctcgcaattg acctaactga gtaagctagc ttgttcgagt attatggcat 2161 tgggaaaact gtttttcttg taccatttgt tgtgcttgta atttactgtg ttttttattc 2221 ggttttcgct atcgaactgt gaaatggaaa tggatggaga agagttaatg aatgatatgg 2281 tccttttgtt cattctcaaa ttaatattat ttgttttttc tcttatttgt tgtgtgttga 2341 atttgaaatt ataagagata tgcaaacatt ttgttttgag taaaaatgtg tcaaatcgtg 2401 gcctctaatg accgaagtta atatgaggag taaaacacta gatccccaaa caagcttgaa 2461 actgaaggcg ggaaacgaca atctgatcat gagcggagaa ttaagggagt cacgttatga 2521 cccccgccga tgacgcggga caagccgttt tacgtttgga actgacagaa ccgcaacgtt 2581 gaaggagcca ctcagccgcg ggtttctgga gtttaatgag ctaagcacat acgtcagaaa 2641 ccattattgc gcgttcaaaa gtcgcctaag gtcactatca gctagcaaat atttcttgtc 2701 aaaaatgctc cactgacgtt ccataaattc ccctcggtat ccaattagag tctcatattc 2761 actctcaatc caaataatct gcaccggatc cgctagagga tctcgacctg caagatcccg 2821 gggggcaatg agatatgaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga 2881 tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg 2941 ctttcagctt cgatatagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg 3001 gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg 3061 aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac 3121 agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg 3181 cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat 3241 tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg 3301 atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc 3361 aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc 3421 acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg 3481 actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga 3541 ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc 3601 ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc 3661 agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa 3721 tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg 3781 tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc 3841 gtccgggatc ttggaggtga tgtaacatga tcacaagctg atcccccgaa tttccccgat 3901 cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg 3961 attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg 4021 acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg 4081 atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg 4141 ttactagatc gggaattgat cccccctcga cagcttgcat gccagcttgg gctgcaggtc 4201 gacgcgtcaa catggtggag cacgacactc tcgtctactc caagaatatc aaagatacag 4261 tctcagaaga ccaaagggct attgagactt ttcaacaaag ggtaatatcg ggaaacctcc 4321 tcggattcca ttgcccagct atctgtcact tcatcaaaag gacagtagaa aaggaaggtg 4381 gcacctacaa atgccatcat tgcgataaag gaaaggctat cgttcaagat gcctctgccg 4441 acagtggtcc caaagatgga cccccaccca cgaggagcat cgtggaaaaa gaagacgttc 4501 caaccacgtc ttcaaagcaa gtggattgat gtgatatctc cactgacgta agggatgacg 4561 cacaatccca ctatccttcg caagaccctt cctctatata aggaagttca tttcatttgg 4621 agaggacacg atatggacaa gaagtactcc attgggctcg atatcggcac aaacagcgtc 4681 ggctgggccg tcattacgga cgagtacaag gtgccgagca aaaaattcaa agttctgggc 4741 aataccgatc gccacagcat aaagaagaac ctcattggcg ccctcctgtt cgactccggg 4801 gagacggccg aagccacgcg gctcaaaaga acagcacggc gcagatatac ccgcagaaag 4861 aatcggatct gctacctgca ggagatcttt agtaatgaga tggctaaggt ggatgactct 4921 ttcttccata ggctggagga gtcctttttg gtggaggagg ataaaaagca cgagcgccac 4981 ccaatctttg gcaatatcgt ggacgaggtg gcgtaccatg aaaagtaccc aaccatatat 5041 catctgagga agaagcttgt agacagtact gataaggctg acttgcggtt gatctatctc 5101 gcgctggcgc atatgatcaa atttcgggga cacttcctca tcgaggggga cctgaaccca 5161 gacaacagcg atgtcgacaa actctttatc caactggttc agacttacaa tcagcttttc 5221 gaagagaacc cgatcaacgc atccggagtt gacgccaaag caatcctgag cgctaggctg 5281 tccaaatccc ggcggctcga aaacctcatc gcacagctcc ctggggagaa gaagaacggc 5341 ctgtttggta atcttatcgc cctgtcactc gggctgaccc ccaactttaa atctaacttc 5401 gacctggccg aagatgccaa gcttcaactg agcaaagaca cctacgatga tgatctcgac 5461 aatctgctgg cccagatcgg cgaccagtac gcagaccttt ttttggcggc aaagaacctg 5521 tcagacgcca ttctgctgag tgatattctg cgagtgaaca cggagatcac caaagctccg 5581 ctgagcgcta gtatgatcaa gcgctatgat gagcaccacc aagacttgac tttgctgaag 5641 gcccttgtca gacagcaact gcctgagaag tacaaggaaa ttttcttcga tcagtctaaa 5701 aatggctacg ccggatacat tgacggcgga gcaagccagg aggaatttta caaatttatt 5761 aagcccatct tggaaaaaat ggacggcacc gaggagctgc tggtaaagct taacagagaa 5821 gatctgttgc gcaaacagcg cactttcgac aatggaagca tcccccacca gattcacctg 5881 ggcgaactgc acgctatcct caggcggcaa gaggatttct accccttttt gaaagataac 5941 agggaaaaga ttgagaaaat cctcacattt cggataccct actatgtagg ccccctcgcc 6001 cggggaaatt ccagattcgc gtggatgact cgcaaatcag aagagaccat cactccctgg 6061 aacttcgagg aagtcgtgga taagggggcc tctgcccagt ccttcatcga aaggatgact 6121 aactttgata aaaatctgcc taacgaaaag gtgcttccta aacactctct gctgtacgag 6181 tacttcacag tttataacga gctcaccaag gtcaaatacg tcacagaagg gatgagaaag 6241 ccagcattcc tgtctggaga gcagaagaaa gctatcgtgg acctcctctt caagacgaac 6301 cggaaagtta ccgtgaaaca gctcaaagaa gactatttca aaaagattga atgtttcgac 6361 tctgttgaaa tcagcggagt ggaggatcgc ttcaacgcat ccctgggaac gtatcacgat 6421 ctcctgaaaa tcattaaaga caaggacttc ctggacaatg aggagaacga ggacattctt 6481 gaggacattg tcctcaccct tacgttgttt gaagataggg agatgattga agaacgcttg 6541 aaaacttacg ctcatctctt cgacgacaaa gtcatgaaac agctcaagag gcgccgatat 6601 acaggatggg ggcggctgtc aagaaaactg atcaatggga tccgagacaa gcagagtgga 6661 aagacaatcc tggattttct taagtccgat ggatttgcca accggaactt catgcagttg 6721 atccatgatg actctctcac ctttaaggag gacatccaga aagcacaagt ttctggccag 6781 ggggacagtc ttcacgagca catcgctaat cttgcaggta gcccagctat caaaaaggga 6841 atactgcaga ccgttaaggt cgtggatgaa ctcgtcaaag taatgggaag gcataagccc 6901 gagaatatcg ttatcgagat ggcccgagag aaccaaacta cccagaaggg acagaagaac 6961 agtagggaaa ggatgaagag gattgaagag ggtataaaag aactggggtc ccaaatcctt 7021 aaggaacacc cagttgaaaa cacccagctt cagaatgaga agctctacct gtactacctg 7081 cagaacggca gggacatgta cgtggatcag gaactggaca tcaatcggct ctccgactac 7141 gacgtggatc atatcgtgcc ccagtctttt ctcaaagatg attctattga taataaagtg 7201 ttgacaagat ccgataaaaa tagagggaag agtgataacg tcccctcaga agaagttgtc 7261 aagaaaatga aaaattattg gcggcagctg ctgaacgcca aactgatcac acaacggaag 7321 ttcgataatc tgactaaggc tgaacgaggt ggcctgtctg agttggataa agccggcttc 7381 atcaaaaggc agcttgttga gacacgccag atcaccaagc acgtggccca aattctcgat 7441 tcacgcatga acaccaagta cgatgaaaat gacaaactga ttcgagaggt gaaagttatt 7501 actctgaagt ctaagctggt ctcagatttc agaaaggact ttcagtttta taaggtgaga 7561 gagatcaaca attaccacca tgcgcatgat gcctacctga atgcagtggt aggcactgca 7621 cttatcaaaa aatatcccaa gcttgaatct gaatttgttt acggagacta taaagtgtac 7681 gatgttagga aaatgatcgc aaagtctgag caggaaatag gcaaggccac cgctaagtac 7741 ttcttttaca gcaatattat gaattttttc aagaccgaga ttacactggc caatggagag 7801 attcggaagc gaccacttat cgaaacaaac ggagaaacag gagaaatcgt gtgggacaag 7861 ggtagggatt tcgcgacagt ccggaaggtc ctgtccatgc cgcaggtgaa catcgttaaa 7921 aagaccgaag tacagaccgg aggcttctcc aaggaaagta tcctcccgaa aaggaacagc 7981 gacaagctga tcgcacgcaa aaaagattgg gaccccaaga aatacggcgg attcgattct 8041 cctacagtcg cttacagtgt actggttgtg gccaaagtgg agaaagggaa gtctaaaaaa 8101 ctcaaaagcg tcaaggaact gctgggcatc acaatcatgg agcgatcaag cttcgaaaaa 8161 aaccccatcg actttctcga ggcgaaagga tataaagagg tcaaaaaaga cctcatcatt 8221 aagcttccca agtactctct ctttgagctt gaaaacggcc ggaaacgaat gctcgctagt 8281 gcgggcgagc tgcagaaagg taacgagctg gcactgccct ctaaatacgt taatttcttg 8341 tatctggcca gccactatga aaagctcaaa gggtctcccg aagataatga gcagaagcag 8401 ctgttcgtgg aacaacacaa acactacctt gatgagatca tcgagcaaat aagcgaattc 8461 tccaaaagag tgatcctcgc cgacgctaac ctcgataagg tgctttctgc ttacaataag 8521 cacagggata agcccatcag ggagcaggca gaaaacatta tccacttgtt tactctgacc 8581 aacttgggcg cgcctgcagc cttcaagtac ttcgacacca ccatagacag aaagcggtac 8641 acctctacaa aggaggtcct ggacgccaca ctgattcatc agtcaattac ggggctctat 8701 gaaacaagaa tcgacctctc tcagctcggt ggagacagca gggctgaccc caagaagaag 8761 aggaaggtgt gatcgatcca ggcctcccag ctttcgtccg tatcatcggt ttcgacaacg 8821 ttcgtcaagt tcaatgcatc agtttcattg cccacacacc agaatcctac taagtttgag 8881 tattatggca ttggaaaagc tgttttcttc tatcatttgt tctgcttgta atttactgtg 8941 ttctttcagt ttttgttttc ggacatcaaa atgcaaatgg atggataaga gttaataaat 9001 gatatggtcc ttttgttcat tctcaaatta ttattatctg ttgtttttac tttaatgggt 9061 tgaatttaag taagaaagga actaacagtg tgatattaag gtgcaatgtt agacatataa 9121 aacagtcttt cacctctctt tggttatgtc ttgaattggt ttgtttcttc acttatctgt 9181 gtaatcaagt ttactatgag tctatgatca agtaattatg caatcaagtt aagtacagta 9241 taggcttttt gtgtcgaggg ggtaccgagt cgaggaattc actggccgtc gttttacaac 9301 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt 9361 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca 9421 gcctgaatgg cgggtaccga gctcgaattc aattcggcgt taattcagta cattaaaaac 9481 gtccgcaatg tgttattaag ttgtctaagc gtcaatttgt ttacaccaca atatatcctg 9541 ccaccagcca gccaacagct ccccgaccgg cagctcggca caaaatcacc actcgataca 9601 ggcagcccat cagtccggga cggcgtcagc gggagagccg ttgtaaggcg gcagactttg 9661 ctcatgttac cgatgctatt cggaagaacg gcaactaagc tgccgggttt gaaacacgga 9721 tgatctcgcg gagggtagca tgttgattgt aacgatgaca gagcgttgct gcctgtgatc 9781 aattcgggca cgaacccagt ggacataagc ctcgttcggt tcgtaagctg taatgcaagt 9841 agcgtaactg ccgtcacgca actggtccag aaccttgacc gaacgcagcg gtggtaacgg 9901 cgcagtggcg gttttcatgg cttcttgtta tgacatgttt ttttggggta cagtctatgc 9961 ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca 10021 gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatc atgggggaag 10081 cggtgatcgc cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc 10141 tcgaaccgac gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc 10201 cacacagtga tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc 10261 gagctttgat caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc 10321 gcgctgtaga agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta 10381 agcgcgaact gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc 10441 cagccacgat cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg 10501 ccttggtagg tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg 10561 aggcgctaaa tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc 10621 gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc 10681 cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca 10741 tacttgaagc tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag 10801 atcagttgga agaatttgtc cactacgtga aaggcgagat caccaaggta gtcggcaaat 10861 aatgtctagc tagaaattcg ttcaagccga cgccgcttcg ccggcgttaa ctcaagcgat 10921 tagatgcact aagcacataa ttgctcacag ccaaactatc aggtcaagtc tgcttttatt 10981 atttttaagc gtgcataata agccctacac aaattgggag atatatcatg catgaccaaa 11041 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 11101 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 11161 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact 11221 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac 11281 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 11341 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 11401 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 11461 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc 11521 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 11581 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 11641 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc 11701 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt 11761 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc 11821 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc 11881 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact 11941 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac 12001 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg 12061 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt 12121 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gggtgccttg atgtgggcgc 12181 cggcggtcga gtggcgacgg cgcggcttgt ccgcgccctg gtagattgcc tggccgtagg 12241 ccagccattt ttgagcggcc agcggccgcg ataggccgac gcgaagcggc ggggcgtagg 12301 gagcgcagcg accgaagggt aggcgctttt tgcagctctt cggctgtgcg ctggccagac 12361 agttatgcac aggccaggcg ggttttaaga gttttaataa gttttaaaga gttttaggcg 12421 gaaaaatcgc cttttttctc ttttatatca gtcacttaca tgtgtgaccg gttcccaatg 12481 tacggctttg ggttcccaat gtacgggttc cggttcccaa tgtacggctt tgggttccca 12541 atgtacgtgc tatccacagg aaagagacct tttcgacctt tttcccctgc tagggcaatt 12601 tgccctagca tctgctccgt acattaggaa ccggcggatg cttcgccctc gatcaggttg 12661 cggtagcgca tgactaggat cgggccagcc tgccccgcct cctccttcaa atcgtactcc 12721 ggcaggtcat ttgacccgat cagcttgcgc acggtgaaac agaacttctt gaactctccg 12781 gcgctgccac tgcgttcgta gatcgtcttg aacaaccatc tggcttctgc cttgcctgcg 12841 gcgcggcgtg ccaggcggta gagaaaacgg ccgatgccgg gatcgatcaa aaagtaatcg 12901 gggtgaaccg tcagcacgtc cgggttcttg ccttctgtga tctcgcggta catccaatca 12961 gctagctcga tctcgatgta ctccggccgc ccggtttcgc tctttacgat cttgtagcgg 13021 ctaatcaagg cttcaccctc ggataccgtc accaggcggc cgttcttggc cttcttcgta 13081 cgctgcatgg caacgtgcgt ggtgtttaac cgaatgcagg tttctaccag gtcgtctttc 13141 tgctttccgc catcggctcg ccggcagaac ttgagtacgt ccgcaacgtg tggacggaac 13201 acgcggccgg gcttgtctcc cttcccttcc cggtatcggt tcatggattc ggttagatgg 13261 gaaaccgcca tcagtaccag gtcgtaatcc cacacactgg ccatgccggc cggccctgcg 13321 gaaacctcta cgtgcccgtc tggaagctcg tagcggatca cctcgccagc tcgtcggtca 13381 cgcttcgaca gacggaaaac ggccacgtcc atgatgctgc gactatcgcg ggtgcccacg 13441 tcatagagca tcggaacgaa aaaatctggt tgctcgtcgc ccttgggcgg cttcctaatc 13501 gacggcgcac cggctgccgg cggttgccgg gattctttgc ggattcgatc agcggccgct 13561 tgccacgatt caccggggcg tgcttctgcc tcgatgcgtt gccgctgggc ggcctgcgcg 13621 gccttcaact tctccaccag gtcatcaccc agcgccgcgc cgatttgtac cgggccggat 13681 ggtttgcgac cgtcacgccg attcctcggg cttgggggtt ccagtgccat tgcagggccg 13741 gcagacaacc cagccgctta cgcctggcca accgcccgtt cctccacaca tggggcattc 13801 cacggcgtcg gtgcctggtt gttcttgatt ttccatgccg cctcctttag ccgctaaaat 13861 tcatctactc atttattcat ttgctcattt actctggtag ctgcgcgatg tattcagata 13921 gcagctcggt aatggtcttg ccttggcgta ccgcgtacat cttcagcttg gtgtgatcct 13981 ccgccggcaa ctgaaagttg acccgcttca tggctggcgt gtctgccagg ctggccaacg 14041 ttgcagcctt gctgctgcgt gcgctcggac ggccggcact tagcgtgttt gtgcttttgc 14101 tcattttctc tttacctcat taactcaaat gagttttgat ttaatttcag cggccagcgc 14161 ctggacctcg cgggcagcgt cgccctcggg ttctgattca agaacggttg tgccggcggc 14221 ggcagtgcct gggtagctca cgcgctgcgt gatacgggac tcaagaatgg gcagctcgta 14281 cccggccagc gcctcggcaa cctcaccgcc gatgcgcgtg cctttgatcg cccgcgacac 14341 gacaaaggcc gcttgtagcc ttccatccgt gacctcaatg cgctgcttaa ccagctccac 14401 caggtcggcg gtggcccata tgtcgtaagg gcttggctgc accggaatca gcacgaagtc 14461 ggctgccttg atcgcggaca cagccaagtc cgccgcctgg ggcgctccgt cgatcactac 14521 gaagtcgcgc cggccgatgg ccttcacgtc gcggtcaatc gtcgggcggt cgatgccgac 14581 aacggttagc ggttgatctt cccgcacggc cgcccaatcg cgggcactgc cctggggatc 14641 ggaatcgact aacagaacat cggccccggc gagttgcagg gcgcgggcta gatgggttgc 14701 gatggtcgtc ttgcctgacc cgcctttctg gttaagtaca gcgataacct tcatgcgttc 14761 cccttgcgta tttgtttatt tactcatcgc atcatatacg cagcgaccgc atgacgcaag 14821 ctgttttact caaatacaca tcaccttttt agacggcggc gctcggtttc ttcagcggcc 14881 aagctggccg gccaggccgc cagcttggca tcagacaaac cggccaggat ttcatgcagc 14941 cgcacggttg agacgtgcgc gggcggctcg aacacgtacc cggccgcgat catctccgcc 15001 tcgatctctt cggtaatgaa aaacggttcg tcctggccgt cctggtgcgg tttcatgctt 15061 gttcctcttg gcgttcattc tcggcggccg ccagggcgtc ggcctcggtc aatgcgtcct 15121 cacggaaggc accgcgccgc ctggcctcgg tgggcgtcac ttcctcgctg cgctcaagtg 15181 cgcggtacag ggtcgagcga tgcacgccaa gcagtgcagc cgcctctttc acggtgcggc 15241 cttcctggtc gatcagctcg cgggcgtgcg cgatctgtgc cggggtgagg gtagggcggg 15301 ggccaaactt cacgcctcgg gccttggcgg cctcgcgccc gctccgggtg cggtcgatga 15361 ttagggaacg ctcgaactcg gcaatgccgg cgaacacggt caacaccatg cggccggccg 15421 gcgtggtggt gtcggcccac ggctctgcca ggctacgcag gcccgcgccg gcctcctgga 15481 tgcgctcggc aatgtccagt aggtcgcggg tgctgcgggc caggcggtct agcctggtca 15541 ctgtcacaac gtcgccaggg cgtaggtggt caagcatcct ggccagctcc gggcggtcgc 15601 gcctggtgcc ggtgatcttc tcggaaaaca gcttggtgca gccggccgcg tgcagttcgg 15661 cccgttggtt ggtcaagtcc tggtcgtcgg tgctgacgcg ggcatagccc agcaggccag 15721 cggcggcgct cttgttcatg gcgtaatgtc tccggttcta gtcgcaagta ttctacttta 15781 tgcgactaaa acacgcgaca agaaaacgcc aggaaaaggg cagggcggca gcctgtcgcg 15841 taacttagga cttgtgcgac atgtcgtttt cagaagacgg ctgcactgaa cgtcagaagc 15901 cgactgcact atagcagcgg aggggttgga tcaaagtact ttaaagtact ttaaagtact 15961 ttaaagtact ttgatcccga ggggaaccct gtggttggca tgcacataca aatggacgaa 16021 cggataaacc ttttcacgcc cttttaaata tccgattatt ctaataaacg ctcttttctc 16081 ttag //