LOCUS Exported 12448 bp ds-DNA circular SYN 21-JAN-2019 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12448) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 21, 2019 from SnapGene Viewer 4.1.1 http://www.snapgene.com COMMENT LOCUS pJK1027_caCas9_f 12448 bp ds-DNA circular 31-MAR-2017 DEFINITION . ACCESSION VERSION SOURCE . ORGANISM . COMMENT pACT1 is 4717-3830.tACT1 is 3642-3249.URA3 locus is 643-704 (piece of URA3 promoter) and again 772-3154 (includes orf).pJK795 insert is 1554-~2795 COMMENT COMMENT ApEinfo:methylated:0 FEATURES Location/Qualifiers source 1..12448 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature complement(132..992) /label=AmpR /note="AmpR" misc_feature complement(1140..1807) /label=pBR322 origin misc_feature 2219..3106 /label=pACT1 /note="pACT1" misc_feature 3237..7376 /label=caCas9 misc_feature complement(7434..7821) /label=ACT1 terminator /note="ACT1 terminator" misc_feature complement(8547..9119) /label=NATr /note="NATr" misc_feature complement(10745..10752) /label=sgRNA 2 misc_feature complement(10753..10831) /label=sgRNA tail misc_feature complement(10753..10831) /label=sgRNA tail(1) misc_feature complement(10840..10846) /label=GAGCAGGTATTAATGACCTGCCTA /note="Geneious type: Editing History Replacement" misc_feature complement(10858..11107) /label=SNR52 promoter misc_feature complement(11113..11612) /label=5' fragment NeutL5 misc_feature complement(11621..12120) /label=3' Neut5L frag ORIGIN 1 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 61 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 121 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 241 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 301 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 421 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 481 taagagaatt atgcagtgct gccataagca tgagtgataa cactgcggcc aacttacttc 541 tgacaacgat cggaggaccg aaggagctaa ccgctttttt tcacaacatg ggggatcatg 601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 661 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 721 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 901 tagttatcta cacgacgggc agtcaggcaa ctatggatga acgaaataga cagatcgctg 961 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 1021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 1081 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 1141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 1201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 1261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 1321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 1381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 1441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 1501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagcattgag 1561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 1621 gaacaggaga gcgcacgagg gagcttccag gggggaacgc ctggtatctt tatagtcctg 1681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggccga 1741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 1801 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 1861 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 1921 aggaagcgta agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 1981 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta 2041 atgtgagtta cctcactcat taggcacccc aggctttaca ctttatgctt ccggctccta 2101 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt 2161 acgccaagct cggaattaac cctcactaaa gggaacaaaa gctgaattgg agggtaccag 2221 agctattaag atcaccagcc tcgtttataa taaacttagt ctaatatttc ttttggacca 2281 aaatagaact gttctcaggt ggcaaaaaac agcttaaagt ctgtgttaaa aaaagatctc 2341 ggctatgcca atcaaaaagg agtgacatta ttatgtaaaa gaagaaatgt tgtaattccg 2401 cgaaacaaca attttaatgc ttactgtgcc gccaccaata tcctatggcc aaggggcctt 2461 taaacatgcc acggaaatat acagtcaaag taaacctact atgtctttag agccttcagg 2521 atcaattgtt tttaaatgaa tgactgcaaa actagaaaag tagcctctta cataagcaag 2581 tcatttgtac accgagagtt tagtaaccag tgaagcaagt atatctggca aacctgaaac 2641 ttatctgtga attataaagc tcataagtaa aaagtgtaga aaagaagata aggcacacca 2701 aatacaaatt tgcaacagta caacgaatat ttcagcgtca aaactagaga ataataaaga 2761 aaacgatctt ttcaaaaaga aaaaaccttt tagttttcct ttgttgttgt tgtgggtgtg 2821 tgctatttat attatatagt ttactcgtaa taccataaaa tattcggttt gattaggtta 2881 ttttaataag ctaatttgtt ttctaatcgt gtaatttatg ctgtgtatat taagtagtgt 2941 gtgtgcactg cccaaaaatg tttgttgttt atagtcggtt aaagagaaaa aagaaaaaaa 3001 gatccataca cacacgttaa ttagttgttc aacgtaatac acttatattt tgttcttatt 3061 tgctttcggt cgctgttctc accaggattt attgccaacg aaacaatttt tttttatata 3121 ttttcagatt tttctttttt tcctttcctt tccttttcta attttcactc ctggttttct 3181 ttctttctta gaaacattat ctcgatatta atattaaaaa aatataatca ttcaaaatgg 3241 ataaaaaata ctccataggt ttggccattg gaaccaattc agttggatgg gctgtaatca 3301 ctgatgaata taaggttcct tctaaaaagt tcaaagtttt aggaaatact gatagacact 3361 ccattaaaaa gaatttaatt ggtgccttgt tgtttgacag tggtgaaaca gctgaagcaa 3421 ctcgtttgaa aagaacagct agaagaagat ataccagaag aaagaacaga atttgttact 3481 tgcaagaaat cttttctaat gaaatggcaa aggttgatga ttcttttttt catagattgg 3541 aagaaagttt cttagttgaa gaagataaga aacatgaaag acatccaatc tttggtaata 3601 ttgtagatga agtggcttac cacgaaaaat atcctacaat ttatcatttg agaaaaaaat 3661 tagttgactc cactgataaa gctgacttaa gacttatata tttggcattg gctcacatga 3721 tcaagttcag aggtcacttc ttaattgaag gtgatttgaa tccagataat tccgatgtcg 3781 acaagttgtt tattcaattg gttcaaactt acaaccaact tttcgaggaa aatccaatca 3841 atgcctcagg tgtggatgct aaagctattt tatctgctag attgtcaaaa agtagacgtt 3901 tggaaaatct tattgcccaa ttgccaggtg aaaaaaaaaa tggattgttt ggaaacttaa 3961 ttgcattatc tttgggtttg accccaaatt tcaaatcaaa ctttgatttg gccgaggatg 4021 ctaaacttca attgtctaag gatacatacg atgacgattt agataatctt cttgcccaaa 4081 ttggtgatca atatgcagat ttattcttgg cagctaagaa cttgagtgat gcaattttgt 4141 taagtgacat tttgagagtc aacacagaaa ttacaaaagc tcctttatcc gcctctatga 4201 ttaaaagata tgatgaacac catcaagatt taaccttgtt gaaagctctt gtacgtcaac 4261 aattaccaga aaagtataaa gaaatattct ttgatcaatc taaaaatggt tacgctggtt 4321 atattgatgg tggtgctagt caagaagaat tttataagtt tatcaagcca attcttgaaa 4381 aaatggatgg tactgaggaa ttgttggtta agttgaatcg agaggattta cttcgtaagc 4441 aaagaacatt tgataacggt tctatccctc atcaaatcca cttaggtgag ttacacgcta 4501 tcttgagacg tcaggaagat ttttaccctt ttttgaaaga caatcgtgaa aaaattgaaa 4561 aaattttgac tttcagaatt ccttattacg tcgggccatt ggctagaggt aattctagat 4621 ttgcatggat gactcgtaaa tctgaagaga ctatcacccc atggaatttt gaggaagttg 4681 ttgataaggg agcctccgct caatcattca tcgagagaat gaccaatttc gacaaaaact 4741 tgccaaatga gaaggtgtta cctaaacatt ctttgttata tgaatatttc actgtttata 4801 atgaattaac aaaagtcaaa tacgtcactg aaggtatgag aaaaccagca ttcttaagtg 4861 gtgaacaaaa gaaagctatc gttgacttgt tattcaaaac taatcgtaaa gtcactgtca 4921 aacaacttaa agaggactat tttaagaaaa tagagtgttt cgattccgtt gaaatcagtg 4981 gggttgagga tagatttaat gctagtttag gaacttatca cgatttgttg aaaattatca 5041 aagacaaaga ttttttggat aatgaagaaa acgaagatat cttggaggac attgtattga 5101 ctttgacttt gtttgaggat agagaaatga tcgaagaacg tttaaaaact tacgctcact 5161 tattcgacga taaagtcatg aaacaattaa aaagaagaag atataccggt tggggaagat 5221 tatctagaaa gttgattaat ggtattagag ataagcaaag tggtaaaacc atcttagatt 5281 tccttaaaag tgatggtttt gcaaacagaa actttatgca gttgattcat gatgatagtt 5341 tgacctttaa ggaagatatt caaaaagctc aggtttctgg acagggtgat tcattgcatg 5401 aacatattgc caacttagca gggtctccag ctatcaaaaa gggaatttta caaactgtta 5461 aagttgttga tgagttggtg aaagttatgg gacgtcataa gcctgaaaat atagttatag 5521 aaatggctag agaaaatcaa acaactcaga aaggacaaaa gaattctcgt gaaagaatga 5581 aaagaataga agaaggaatt aaagagttgg gttctcaaat cttaaaagaa cacccagtcg 5641 aaaacaccca gttacagaac gagaagcttt acttgtacta tttgcaaaac gggcgtgaca 5701 tgtatgtaga tcaagagttg gatatcaata gactttcaga ttacgatgtc gatcacatag 5761 ttcctcagtc atttttaaaa gatgattcca ttgacaacaa agttttaaca agatctgata 5821 aggctagagg taaatcagac aatgtgccat cagaagaagt ggttaagaaa atgaagaact 5881 attggagaca attgttaaat gcaaagttga ttactcagag aaaatttgac aacttaacta 5941 aagctgaaag aggaggatta tccgaattag ataaagctgg attcataaaa cgtcagttgg 6001 ttgaaacaag acaaattact aaacatgtag cccaaatttt agattcccga atgaatacta 6061 aatatgatga aaatgacaaa ttaatcagag aggtcaaagt gatcacatta aagtcaaagt 6121 tggtgtcaga tttccgaaag gattttcaat tctacaaggt gagagagatc aacaattatc 6181 atcatgctca cgacgcatat ttgaatgcag tggtaggaac tgccttaata aagaagtatc 6241 ctaagttaga atccgaattc gtgtatggtg attataaagt ctacgatgtc agaaagatga 6301 tagcaaaatc agaacaagaa attgggaaag ccaccgcaaa gtatttcttt tactccaaca 6361 tcatgaattt ctttaagacc gaaatcacct tggctaacgg ggaaatcaga aaacgtcctt 6421 tgattgaaac aaatggagaa accggtgaaa ttgtgtggga taaggggaga gattttgcta 6481 ctgttagaaa agttttatca atgcctcaag ttaatattgt aaagaagact gaagttcaaa 6541 ctggtggttt ctctaaggaa tccattcttc caaaacgaaa ctcagacaag ttgatcgccc 6601 gaaaaaaaga ttgggatcct aagaaatacg gaggttttga ttctcctact gtcgcatatt 6661 ctgtcttggt cgtggctaaa gttgaaaagg gtaaatccaa aaaattgaaa tctgtgaaag 6721 aattgttagg tattaccatc atggagcgtt catcctttga gaaaaaccca attgactttc 6781 tcgaagcgaa gggctataaa gaagtgaaga aagatttgat tattaagttg ccaaagtatt 6841 ctctctttga acttgaaaat gggagaaaga gaatgttggc ttcagctgga gaattgcaaa 6901 aaggaaacga gttagctttg ccttcaaagt acgttaattt cttatactta gcatctcatt 6961 atgaaaagtt gaaggggtcc cctgaagaca atgagcaaaa gcaattattc gtagaacaac 7021 ataaacacta tttggatgag ataattgaac aaatctccga gttttctaaa agagttattt 7081 tggctgatgc aaatttggat aaagttttat ctgcttataa taagcatcgt gacaaaccaa 7141 ttcgagaaca agctgaaaat attatccact tgttcacatt gactaacttg ggagctccag 7201 ccgcgtttaa gtactttgac accaccattg accgtaagag atatacttca acaaaagaag 7261 tgttagacgc cacattgatt catcagtcca taactggttt atatgaaact agaatcgatt 7321 tgtcacaatt ggggggtgat agccgagcgg atcctaagaa aaaacgaaaa gtctgactcg 7381 aggtcgacgg tatcgataag cttgatatcg aattcctgca gcccggggga tccgagtgaa 7441 attctggaaa tctggaaatc tggttttgta ttcttgttat tcttcttttt gttattacat 7501 atataacttg ttactttttt aaaaaaatct ttgtttattt tataaatata taaaactaaa 7561 tttaagaaaa agagaaaaat gttttatttg agagattgat attttacttg aatttagctt 7621 agcttttata aagtattatt atgtaaaaaa acaaaacaaa tatacattaa aaagttaaga 7681 ctataaaata gccacccaag gcatttctat atcttgttgt tgttgttttc atcttctgta 7741 tcagaggaac ttattttatt attttcgtca cgggtatttt ctcttgtttg atgattcatc 7801 ccattcattc catcataaaa ttctagaagg accacctttg attgtaaata gtaataatta 7861 ccacccttat ttaattattt atttaactta tttatttatt tattctacat atatacaaat 7921 ctaataaagt gaaaatctcc cccttcacat ttataattgg ccagtctttt tcaaataagc 7981 attccaacca gcatttctat accttttacc ttcaatatct ggatctcttc ctttaccaaa 8041 caatcctcta ccaacaatga taatatcagt tccagtgcta acaacttcat caacagttct 8101 atattgttgt cctaatccat cacctttatc atctaatcca actccaggtg tcataataag 8161 ccaatcaaat ccttcttctt ggccacccat atcacgttgg gcaataaatc caataacaaa 8221 ttccttatcg gatttagcaa tttcaacagt tttttgagaa tattctccat agtaccgaat 8281 tcgagctcgt ttaaacgtcg acactggatg gcggcgttag tatcgaatcg acagcagtat 8341 agcgaccagc attcacatac gattgacgca tgatattact ttctgcgcac ttaacttcgc 8401 atctgggcag atgatgtcga ggcgaaaaaa aatataaatc acgctaacat ttgattaaaa 8461 tagaacaact acaatataaa aaaactatac aaatgacaag ttcttgaaaa caagaatctt 8521 tttattgtca gtactgactc gagttattat ggacatggca tagacatata caaagcttgt 8581 tcaccatcgg aagcagtacc atcgtataaa gcagtatcca aaccacacaa agtgaaaccc 8641 attcttctat aagcatgaat agctggagca ttaacattgg taacttccaa ccacaaatga 8701 ccagcacctc tttctctggc gaattcagta gccaaaccca tcaaagctct accaacacca 8761 tgacctctat gttctggagc aacttcaata tcttcaacag tcaatcttct gttccaacca 8821 gaataagaaa caacaacgaa accagccaaa tcaccatcat caccataagc aacgaaagtt 8881 ctagaatctg gatcaccatc ttcaccagca tcggattcat catcggattc atcatctggg 8941 aaaaccttag tcaatggtgg atcaactgga acttctctca aagtgaaacc atcaccagta 9001 gcagtaactc taaaaacagt atcggtagtg aaagaaccat ccaaagcttc aatagcttca 9061 gcatcacctg gaacagaagt tctgtatcta taagcagtat catccaaagt agtagacata 9121 attgtaggat ccggttgttt atgttcggat gtgatgtgag aactgtatcc tagcaagatt 9181 ttaaaaggaa gtatatgaaa gaagaacctc agtggcaaat cctaaccttt tatatttctc 9241 tacaggggcg cggcgtgggg acaattcaac gcgtctgtga ggggagcgtt tccctgctcg 9301 caggtctgca gcgaggagcc gtaatttttg cttcgcgccg tgcggccatc aaaatgtatg 9361 gatgcaaatg attatacatg gggatgtatg ggctaaatgt acgggcgaca gtcacatcat 9421 gcccctgagc tgcgcacgtc aagactgtca aggagggtat tctgggcctc catgtcgctg 9481 gccgggtgac ccggcgggga cgaggcaagc ttgattgcta atgatcccac tgatgataat 9541 tcagctaaca tcaataaccc tcttggctct tggttggtgg tggtttcttt agctccctgt 9601 tttaatcctt caaccactcc attcccagtg acaccatgag cattggtaat atctgcccaa 9661 ctactaattt tataaactcc accaatatat tgtttcttta cggtattacc aatatcagca 9721 aattttctat cttcaaaaat cataaattga tgtttacgtg aaagttctaa taatggttca 9781 atagtggatt cataggaaaa atcattgatt atatcaatat gagtcttgat taagcataca 9841 taaggaccta atttatcaat taattcaagg aattccttag tggtatcaac gtcaattgat 9901 gcacataaat tggttttctt cagttccatt aatcgaaata atcgctgtgc tactggtgag 9961 gcatgagttt ctgctctctc actataggtc ttagtgttga ctgtcatatc ttttgtttgc 10021 tatttctatt tctatttctt tcttcaactg taaaaggggc ggtatataaa ccagtgtgta 10081 tggggttgtt gctcaggaat tgatcaatgt ttaaaagaat ggttgaaatc atcattgaag 10141 aacaaaaaaa aagatttttt ctttcattat gttatctgta taatatgcag gaaataagat 10201 tgcaaaaaaa tagccgtttt tttggatact atcaaacaag aggaatgact aatacagaat 10261 tagtgttacg aatcaatggc actacagcaa ctttcaacaa aataatctac gttcaaaata 10321 taatatatac tattaaataa aatcaaataa taaaataatt ttaaagaaag tagtaccagc 10381 tctttttttt gtttccgttt ataccatcca aatcaattcc tattagtcgc cctatagtga 10441 gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt 10501 tacccaactt aatcgccttg cagcacatcc ccccttcgcc agctggcgta atagcgaaga 10561 ggcccgcacc gatcgccctt cccaacagtt gcgtagcctg aatggcgaat gggacgcgcc 10621 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 10681 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 10741 cggcaaaaaa aagcaccacc gactcggtgc cactttttca agttgataac ggactagcct 10801 tattttaact tgctatttct agctctaaaa ctgaagagcc attaatgctc ttcctcgaaa 10861 ttaaaaatag tttacgcaag tctcgaactt ccaccacaat ttaattgtaa gaacgctgcg 10921 aaaacatatt ctccaatcct attgggccga agtttcagct atgccaaagg catcaaattt 10981 ttttttataa aattttccac ttttccatac cagtgaaatt atcacgtgat ggagtgagta 11041 tctataatat ttgattgttt ctttgatgag gcaatactcg agtatagtta ttgaccaatg 11101 acttttaccg gctaaacaag tggtattcaa gcacaattca gttttctgga tctctaataa 11161 ttgcaattgc aattgcttca catactctgt ttataattca gtttctttct cttgatgtta 11221 ctgatatatt aagttaataa ataccaataa ctttttttca aaaatcttct attttccttt 11281 caattgatta tttaaacacg aatcgttaat aagctgtgat tgctttttct ttttcttttt 11341 cttttctttg ttcaagtttg aaaaaagtaa aaaaaaaggt ttctcttgtt cgatgtagaa 11401 ttcagtgggt tatagttgta atgatatatg gaataaaaag aaagaaagaa agagaagaga 11461 aagagaagaa gagaagaatt aagtaaacgt gtattagaat ttaaaaatta aaattcatga 11521 taatctttaa taataattaa ctaagaatca caaaattgaa aatttgaaag aaaaaagatc 11581 actccaaggt cgtgtcaagt gactagaaat tcttaattaa aatagttata acaaaaaaag 11641 aaatttgtat ataacaatta agtaaaagaa aaaaaagcaa ttcgctttgt ataataatac 11701 ccagaaaaat atattcatat attcataaat caaacatgaa atgaagaatg ctgaatcact 11761 tgataggatt tagttccatt atggtactaa gaagtaatat tacttctatg aaaaggttga 11821 atccgagatc aaaattcatc ttttcatata ctctggagaa aaaaaaaaaa gtaattgtag 11881 taagaatgac aagtatcagt tgagaaaaaa agtaacaaag agaaagctcg gaggaggctc 11941 cccaaaggtt ttatcaccaa tggtggtacc actaacccaa gaacagaaaa agcattgata 12001 ttttgaaaaa atacaatact catcttttaa cccgaacctt ttcttgtcct ttaaaacttt 12061 tatctcatat ctgtccaaag tcatattgtc ttactacttt ctctccttca tcgtccttcc 12121 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta 12181 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc 12241 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg 12301 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt 12361 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat 12421 tttaacaaaa tattaacgtt tacaattt //