LOCUS Exported 7614 bp ds-DNA circular SYN 26-SEP-2017 DEFINITION Human expression plasmid for SpCas9 eSpCas9(1.1)-HF1 variant: CMV-T7-hSpCas9-eSpCas9(1.1)-HF1(N497A, R661A, Q695A, K848A, Q926A, K1003A, R1060A)-NLS(SV40)-3xFLAG . ACCESSION urn.local...187-7xx66k5 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7614) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 26, 2017 from SnapGene 4.0.5 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7614 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 287..356 promoter 377..396 misc_feature 403..412 /label=Geneious type: motif /note="Geneious type: motif" misc_feature 409..4512 /label=Geneious type: CDS /note="Geneious type: CDS" misc_feature 1833 /label=Geneious type: Editing History Replacement /note="Geneious type: Editing History Replacement" misc_binding 1897..1899 misc_feature 1897..1898 /label=Geneious type: Editing History Replacement /note="Geneious type: Editing History Replacement" misc_binding 2389..2391 misc_feature 2389..2391 /label=Geneious type: Editing History Replacement /note="Geneious type: Editing History Replacement" misc_binding 2491..2493 misc_feature 2491..2493 /label=Geneious type: Editing History Replacement /note="Geneious type: Editing History Replacement" misc_binding 2950..2952 misc_feature 2950..2952 /label=Geneious type: Editing History Replacement /note="Geneious type: Editing History Replacement" misc_binding 3184..3186 misc_feature 3184..3186 /label=Geneious type: Editing History Replacement /note="Geneious type: Editing History Replacement" misc_feature 3204 /label=Geneious type: Editing History Replacement /note="Geneious type: Editing History Replacement" misc_binding 3415..3417 misc_feature 3415..3417 /label=Geneious type: Editing History Replacement /note="Geneious type: Editing History Replacement" misc_binding 3586..3588 misc_feature 3586..3588 /label=Geneious type: Editing History Replacement /note="Geneious type: Editing History Replacement" misc_feature 4519..4539 /label=Geneious type: ORF /note="Geneious type: ORF" misc_feature 4546..4611 /label=Geneious type: ORF /note="Geneious type: ORF" terminator 4672..4899 promoter complement(5025..5048) terminator 5158..5289 misc_feature complement(5357..5976) /label=Geneious type: origin of replication /note="Geneious type: origin of replication" gene complement(6131..6991) /gene="Ampicillin" /label=Ampicillin /note="Geneious type: CDS" promoter complement(6992..7096) misc_feature 7380..129 /label=Geneious type: promoter /note="Geneious type: promoter" promoter complement(7429..7457) ORIGIN 1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg 61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg 121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact 181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa 241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta 301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct 361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat ggataaaaag 421 tattctattg gtttagacat cggcactaat tccgttggat gggctgtcat aaccgatgaa 481 tacaaagtac cttcaaagaa atttaaggtg ttggggaaca cagaccgtca ttcgattaaa 541 aagaatctta tcggtgccct cctattcgat agtggcgaaa cggcagaggc gactcgcctg 601 aaacgaaccg ctcggagaag gtatacacgt cgcaagaacc gaatatgtta cttacaagaa 661 atttttagca atgagatggc caaagttgac gattctttct ttcaccgttt ggaagagtcc 721 ttccttgtcg aagaggacaa gaaacatgaa cggcacccca tctttggaaa catagtagat 781 gaggtggcat atcatgaaaa gtacccaacg atttatcacc tcagaaaaaa gctagttgac 841 tcaactgata aagcggacct gaggttaatc tacttggctc ttgcccatat gataaagttc 901 cgtgggcact ttctcattga gggtgatcta aatccggaca actcggatgt cgacaaactg 961 ttcatccagt tagtacaaac ctataatcag ttgtttgaag agaaccctat aaatgcaagt 1021 ggcgtggatg cgaaggctat tcttagcgcc cgcctctcta aatcccgacg gctagaaaac 1081 ctgatcgcac aattacccgg agagaagaaa aatgggttgt tcggtaacct tatagcgctc 1141 tcactaggcc tgacaccaaa ttttaagtcg aacttcgact tagctgaaga tgccaaattg 1201 cagcttagta aggacacgta cgatgacgat ctcgacaatc tactggcaca aattggagat 1261 cagtatgcgg acttattttt ggctgccaaa aaccttagcg atgcaatcct cctatctgac 1321 atactgagag ttaatactga gattaccaag gcgccgttat ccgcttcaat gatcaaaagg 1381 tacgatgaac atcaccaaga cttgacactt ctcaaggccc tagtccgtca gcaactgcct 1441 gagaaatata aggaaatatt ctttgatcag tcgaaaaacg ggtacgcagg ttatattgac 1501 ggcggagcga gtcaagagga attctacaag tttatcaaac ccatattaga gaagatggat 1561 gggacggaag agttgcttgt aaaactcaat cgcgaagatc tactgcgaaa gcagcggact 1621 ttcgacaacg gtagcattcc acatcaaatc cacttaggcg aattgcatgc tatacttaga 1681 aggcaggagg atttttatcc gttcctcaaa gacaatcgtg aaaagattga gaaaatccta 1741 acctttcgca taccttacta tgtgggaccc ctggcccgag ggaactctcg gttcgcatgg 1801 atgacaagaa agtccgaaga aacgattact ccctggaatt ttgaggaagt tgtcgataaa 1861 ggtgcgtcag ctcaatcgtt catcgagagg atgaccgcct ttgacaagaa tttaccgaac 1921 gaaaaagtat tgcctaagca cagtttactt tacgagtatt tcacagtgta caatgaactc 1981 acgaaagtta agtatgtcac tgagggcatg cgtaaacccg cctttctaag cggagaacag 2041 aagaaagcaa tagtagatct gttattcaag accaaccgca aagtgacagt taagcaattg 2101 aaagaggact actttaagaa aattgaatgc ttcgattctg tcgagatctc cggggtagaa 2161 gatcgattta atgcgtcact tggtacgtat catgacctcc taaagataat taaagataag 2221 gacttcctgg ataacgaaga gaatgaagat atcttagaag atatagtgtt gactcttacc 2281 ctctttgaag atcgggaaat gattgaggaa agactaaaaa catacgctca cctgttcgac 2341 gataaggtta tgaaacagtt aaagaggcgt cgctatacgg gctggggagc cttgtcgcgg 2401 aaacttatca acgggataag agacaagcaa agtggtaaaa ctattctcga ttttctaaag 2461 agcgacggct tcgccaatag gaactttatg gccctgatcc atgatgactc tttaaccttc 2521 aaagaggata tacaaaaggc acaggtttcc ggacaagggg actcattgca cgaacatatt 2581 gcgaatcttg ctggttcgcc agccatcaaa aagggcatac tccagacagt caaagtagtg 2641 gatgagctag ttaaggtcat gggacgtcac aaaccggaaa acattgtaat cgagatggca 2701 cgcgaaaatc aaacgactca gaaggggcaa aaaaacagtc gagagcggat gaagagaata 2761 gaagagggta ttaaagaact gggcagccag atcttaaagg agcatcctgt ggaaaatacc 2821 caattgcaga acgagaaact ttacctctat tacctacaaa atggaaggga catgtatgtt 2881 gatcaggaac tggacataaa ccgtttatct gattacgacg tcgatcacat tgtaccccaa 2941 tcctttttgg ccgacgattc aatcgacaat aaagtgctta cacgctcgga taagaaccga 3001 gggaaaagtg acaatgttcc aagcgaggaa gtcgtaaaga aaatgaagaa ctattggcgg 3061 cagctcctaa atgcgaaact gataacgcaa agaaagttcg ataacttaac taaagctgag 3121 aggggtggct tgtctgaact tgacaaggcc ggatttatta aacgtcagct cgtggaaacc 3181 cgcgccatca caaagcatgt tgcgcagata ctagattccc gaatgaatac gaaatacgac 3241 gagaacgata agctgattcg ggaagtcaaa gtaatcactt taaagtcaaa attggtgtcg 3301 gacttcagaa aggattttca attctataaa gttagggaga taaataacta ccaccatgcg 3361 cacgacgctt atcttaatgc cgtcgtaggg accgcactca ttaagaaata cccggcccta 3421 gaaagtgagt ttgtgtatgg tgattacaaa gtttatgacg tccgtaagat gatcgcgaaa 3481 agcgaacagg agataggcaa ggctacagcc aaatacttct tttattctaa cattatgaat 3541 ttctttaaga cggaaatcac tctggcaaac ggagagatac gcaaagcccc tttaattgaa 3601 accaatgggg agacaggtga aatcgtatgg gataagggcc gggacttcgc gacggtgaga 3661 aaagttttgt ccatgcccca agtcaacata gtaaagaaaa ctgaggtgca gaccggaggg 3721 ttttcaaagg aatcgattct tccaaaaagg aatagtgata agctcatcgc tcgtaaaaag 3781 gactgggacc cgaaaaagta cggtggcttc gatagcccta cagttgccta ttctgtccta 3841 gtagtggcaa aagttgagaa gggaaaatcc aagaaactga agtcagtcaa agaattattg 3901 gggataacga ttatggagcg ctcgtctttt gaaaagaacc ccatcgactt ccttgaggcg 3961 aaaggttaca aggaagtaaa aaaggatctc ataattaaac taccaaagta tagtctgttt 4021 gagttagaaa atggccgaaa acggatgttg gctagcgccg gagagcttca aaaggggaac 4081 gaactcgcac taccgtctaa atacgtgaat ttcctgtatt tagcgtccca ttacgagaag 4141 ttgaaaggtt cacctgaaga taacgaacag aagcaacttt ttgttgagca gcacaaacat 4201 tatctcgacg aaatcataga gcaaatttcg gaattcagta agagagtcat cctagctgat 4261 gccaatctgg acaaagtatt aagcgcatac aacaagcaca gggataaacc catacgtgag 4321 caggcggaaa atattatcca tttgtttact cttaccaacc tcggcgctcc agccgcattc 4381 aagtattttg acacaacgat agatcgcaaa cgatacactt ctaccaagga ggtgctagac 4441 gcgacactga ttcaccaatc catcacggga ttatatgaaa ctcggataga tttgtcacag 4501 cttgggggtg acggatcccc caagaagaag aggaaagtct cgagcgacta caaagaccat 4561 gacggtgatt ataaagatca tgacatcgat tacaaggatg acgatgacaa ggctgcagga 4621 tgaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc 4681 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag 4741 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta 4801 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag 4861 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca 4921 gctggggctc gataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 4981 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 5041 agtgtaaagc ctagggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 5101 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 5161 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 5221 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 5281 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 5341 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 5401 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 5461 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 5521 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 5581 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 5641 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 5701 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 5761 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 5821 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 5881 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 5941 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 6001 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 6061 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 6121 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 6181 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 6241 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 6301 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 6361 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 6421 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 6481 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 6541 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 6601 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 6661 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 6721 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 6781 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 6841 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 6901 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 6961 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 7021 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 7081 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag 7141 atcgatctcc cgatccccta gggtcgactc tcagtacaat ctgctctgat gccgcatagt 7201 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa 7261 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta 7321 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga 7381 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc 7441 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat 7501 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc 7561 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatc //