LOCUS Exported 9178 bp ds-DNA circular SYN 18-MAY-2020 DEFINITION . ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9178) AUTHORS . TITLE Direct Submission JOURNAL Exported Monday, May 18, 2020 from SnapGene Viewer 5.1.2 https://www.snapgene.com FEATURES Location/Qualifiers source 1..9178 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..241 /label=U6 promoter /note="color: #c6c9d1" misc_feature 246..274 /label=567/568 /note="color: #ff9ccd" misc_feature 251..270 /label=gRNA sequence /note="color: #33cccc" misc_RNA 271..346 /label=gRNA scaffold /note="color: #85dae9; direction: RIGHT" promoter 730..1007 /label=chicken beta-actin promoter /note="color: #c6c9d1" CDS 1257..1322 /label=3xFLAG /note="color: #ff9ccd; direction: RIGHT" CDS 1329..1349 /label=SV40 NLS /note="color: #993366; direction: RIGHT" CDS 1374..5474 /label=Cas9 /note="color: #993366; direction: RIGHT" CDS 5475..5522 /label=nucleoplasmin NLS /note="color: #993366; direction: RIGHT" CDS 5538..5591 /label=T2A /note="color: #ff9ccd; direction: RIGHT" CDS 5592..6191 /label=PuroR /note="color: #b7e6d7; direction: RIGHT" polyA_signal 6225..6432 /label=bGH poly(A) signal /note="color: #b4abac" repeat_region 6441..6581 /label=AAV2 ITR /note="color: #f8d3a9" rep_origin 6656..7111 /label=f1 ori /note="color: #ffef86" promoter 7393..7497 /label=AmpR promoter /note="color: #c6c9d1" CDS 7498..7566 /label=AmpR /note="color: #b7e6d7; direction: RIGHT" CDS 7567..8358 /label=AmpR /note="color: #b7e6d7; direction: RIGHT" rep_origin 8529..9117 /label=ori /note="color: #ffef86" ORIGIN 1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag 61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga 121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat 181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga 241 cgaaacaccg tgagtagcca ttgtccacgc gttttagagc tagaaatagc aagttaaaat 301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga 361 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca 421 gacaaatggc tctagaggta cccgttacat aacttacggt aaatggcccg cctggctgac 481 cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact ttccattgac 541 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata 601 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattgtgccc 661 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta 721 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc 781 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg 841 gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc 901 ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga 961 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcga 1021 cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga 1081 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat 1141 tagctgagca agaggtaagg gtttaaggga tggttggttg gtggggtatt aatgtttaat 1201 tacctggagc acctgcctga aatcactttt tttcaggttg gaccggtgcc accatggact 1261 ataaggacca cgacggagac tacaaggatc atgatattga ttacaaagac gatgacgata 1321 agatggcccc aaagaagaag cggaaggtcg gtatccacgg agtcccagca gccgacaaga 1381 agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg atcaccgacg 1441 agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca 1501 agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc 1561 tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag 1621 agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt 1681 ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg 1741 acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg 1801 acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt 1861 tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc 1921 tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca 1981 gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa 2041 atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac ctgattgccc 2101 tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac 2161 tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg 2221 accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg 2281 acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga 2341 gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc 2401 ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg 2461 acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg 2521 acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga 2581 ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc 2641 ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc 2701 tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct 2761 ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca 2821 agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca 2881 acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc 2941 tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc 3001 agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc 3061 tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg 3121 aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca 3181 aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga 3241 cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg 3301 acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc 3361 ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga 3421 agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct 3481 ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca 3541 ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg 3601 tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg 3661 ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga 3721 tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca 3781 cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg 3841 tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc 3901 agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc 3961 ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc 4021 ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg 4081 agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa 4141 cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg 4201 acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt 4261 ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg 4321 cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc 4381 tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca 4441 agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga 4501 actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg 4561 agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc 4621 ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg 4681 gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga 4741 aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc 4801 tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc 4861 tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag 4921 ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt 4981 tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa 5041 acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga 5101 agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc 5161 actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg 5221 acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag 5281 agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct 5341 tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg 5401 acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc 5461 agctgggagg cgacaaaagg ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa 5521 aggaattcgg cagtggagag ggcagaggaa gtctgctaac atgcggtgac gtcgaggaga 5581 atcctggccc aatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc 5641 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg 5701 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg 5761 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga 5821 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg 5881 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc 5941 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggagt ctcgcccgac caccagggca 6001 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc 6061 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca 6121 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc 6181 ccggtgcctg agaattctaa ctagagctcg ctgatcagcc tcgactgtgc cttctagttg 6241 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc 6301 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc 6361 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag agaatagcag 6421 gcatgctggg gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg 6481 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg 6541 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt 6601 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg 6661 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 6721 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 6781 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 6841 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg 6901 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 6961 ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg 7021 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 7081 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct 7141 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg 7201 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg 7261 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc 7321 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt 7381 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 7441 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg 7501 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt 7561 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga 7621 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa 7681 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt 7741 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt 7801 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc 7861 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga 7921 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat 7981 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct 8041 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc 8101 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg 8161 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tggaagccgc 8221 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg 8281 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca 8341 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta 8401 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 8461 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 8521 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 8581 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 8641 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc 8701 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 8761 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 8821 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 8881 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 8941 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 9001 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 9061 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 9121 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt //