LOCUS Exported 10723 bp ds-DNA circular SYN 20-SEP-2017 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Ezh2[SET]-iCas9v2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10723) AUTHORS . TITLE Direct Submission JOURNAL Exported Wednesday, Sep 20, 2017 from SnapGene Viewer 3.3.4 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10723 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 196..405 /label=CMV promoter misc_feature 529..552 /label=SV40 NLS1 misc_feature 568..633 /label=3XFLAG misc_feature 664..1461 /label=mEzh2-SET misc_feature 1522..5625 /label=dCas9_D10A_H840A misc_feature 5701..5748 /label=Nucleoplasmin NLS /note="note=""direct strand translation""translation=""KRPAATKKAGQAKKKK""" misc_feature 5767..5823 /label=P2A misc_feature 6624..6839 /label=SV40 enhancer-pBabe misc_feature 6803..6880 /label=SV40 ORI CDS 7022..7813 /label=Kan/neoR misc_binding complement(8181..8203) /label=LacO promoter complement(8208..8237) /label=lac rep_origin complement(8543..9171) /direction=LEFT /label=ColE1 origin CDS complement(9323..9982) /label=AmpR promoter complement(10222..10250) /label=Amp prom misc_feature 10631..195 /label=CAG enhancer ORIGIN 1 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca 61 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg 121 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc 181 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt 241 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt 301 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg 361 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg 421 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg 481 gaccgatcca gcctccggac tctagaggat cgaacccttg ccaccatgcc caagaagaag 541 aggaaggtgg gaggctccgg aggaagcgac tataaggacc acgacggaga ctacaaggat 601 catgatattg attacaaaga cgatgacgat aagggtggcg ggtccggcgg tggatccggt 661 accatgactg aggatgtaga cactcctcca agaaagaaga aaaggaaaca tcggttgtgg 721 gctgcacact gcagaaagat acaactgaaa aaggacggct cctctaacca tgtttacaac 781 tatcaaccct gtgaccatcc acggcagcct tgtgacagtt cgtgcccttg tgtgatagca 841 caaaattttt gtgaaaagtt ttgtcaatgt agttcagagt gtcaaaaccg ctttcctgga 901 tgtcggtgca aagcacaatg caacaccaaa cagtgtccat gctacctggc tgtccgagag 961 tgtgaccctg acctctgtct cacgtgtgga gctgctgacc attgggacag taaaaatgta 1021 tcctgtaaga actgtagcat tcagcggggc tctaaaaagc acttactgct ggcaccgtct 1081 gatgtggcag gctggggcat ctttatcaaa gatcctgtac agaaaaatga attcatctca 1141 gaatactgtg gggagattat ttctcaggat gaagcagaca gaagaggaaa agtgtatgac 1201 aaatacatgt gcagctttct gttcaacttg aacaatgatt ttgtggtgga tgcaacccga 1261 aagggcaaca aaattcgttt tgctaatcat tcagtaaatc caaactgcta tgcaaaagtt 1321 atgatggtta atggtgacca caggataggc atctttgcta agagggctat ccagactggt 1381 gaagagttgt tttttgatta cagatacagc caggctgatg ccctgaagta tgtgggcatc 1441 gaacgagaaa tggaaatccc tagtaccgga gggagtggtg gaagcggcgg ttctggtggc 1501 tcaggcggca gcggccggcc aatggacaag aagtactcca ttgggctcgc tatcggcaca 1561 aacagcgtcg gctgggccgt cattacggac gagtacaagg tgccgagcaa aaaattcaaa 1621 gttctgggca ataccgatcg ccacagcata aagaagaacc tcattggcgc cctcctgttc 1681 gactccgggg agacggccga agccacgcgg ctcaaaagaa cagcacggcg cagatatacc 1741 cgcagaaaga atcggatctg ctacctgcag gagatcttta gtaatgagat ggctaaggtg 1801 gatgactctt tcttccatag gctggaggag tcctttttgg tggaggagga taaaaagcac 1861 gagcgccacc caatctttgg caatatcgtg gacgaggtgg cgtaccatga aaagtaccca 1921 accatatatc atctgaggaa gaagcttgta gacagtactg ataaggctga cttgcggttg 1981 atctatctcg cgctggcgca tatgatcaaa tttcggggac acttcctcat cgagggggac 2041 ctgaacccag acaacagcga tgtcgacaaa ctctttatcc aactggttca gacttacaat 2101 cagcttttcg aagagaaccc gatcaacgca tccggagttg acgccaaagc aatcctgagc 2161 gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg cacagctccc tggggagaag 2221 aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc caactttaaa 2281 tctaacttcg acctggccga agatgccaag cttcaactga gcaaagacac ctacgatgat 2341 gatctcgaca atctgctggc ccagatcggc gaccagtacg cagacctttt tttggcggca 2401 aagaacctgt cagacgccat tctgctgagt gatattctgc gagtgaacac ggagatcacc 2461 aaagctccgc tgagcgctag tatgatcaag cgctatgatg agcaccacca agacttgact 2521 ttgctgaagg cccttgtcag acagcaactg cctgagaagt acaaggaaat tttcttcgat 2581 cagtctaaaa atggctacgc cggatacatt gacggcggag caagccagga ggaattttac 2641 aaatttatta agcccatctt ggaaaaaatg gacggcaccg aggagctgct ggtaaagctt 2701 aacagagaag atctgttgcg caaacagcgc actttcgaca atggaagcat cccccaccag 2761 attcacctgg gcgaactgca cgctatcctc aggcggcaag aggatttcta cccctttttg 2821 aaagataaca gggaaaagat tgagaaaatc ctcacatttc ggatacccta ctatgtaggc 2881 cccctcgccc ggggaaattc cagattcgcg tggatgactc gcaaatcaga agagaccatc 2941 actccctgga acttcgagga agtcgtggat aagggggcct ctgcccagtc cttcatcgaa 3001 aggatgacta actttgataa aaatctgcct aacgaaaagg tgcttcctaa acactctctg 3061 ctgtacgagt acttcacagt ttataacgag ctcaccaagg tcaaatacgt cacagaaggg 3121 atgagaaagc cagcattcct gtctggagag cagaagaaag ctatcgtgga cctcctcttc 3181 aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag actatttcaa aaagattgaa 3241 tgtttcgact ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc cctgggaacg 3301 tatcacgatc tcctgaaaat cattaaagac aaggacttcc tggacaatga ggagaacgag 3361 gacattcttg aggacattgt cctcaccctt acgttgtttg aagataggga gatgattgaa 3421 gaacgcttga aaacttacgc tcatctcttc gacgacaaag tcatgaaaca gctcaagagg 3481 cgccgatata caggatgggg gcggctgtca agaaaactga tcaatgggat ccgagacaag 3541 cagagtggaa agacaatcct ggattttctt aagtccgatg gatttgccaa ccggaacttc 3601 atgcagttga tccatgatga ctctctcacc tttaaggagg acatccagaa agcacaagtt 3661 tctggccagg gggacagtct tcacgagcac atcgctaatc ttgcaggtag cccagctatc 3721 aaaaagggaa tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt aatgggaagg 3781 cataagcccg agaatatcgt tatcgagatg gcccgagaga accaaactac ccagaaggga 3841 cagaagaaca gtagggaaag gatgaagagg attgaagagg gtataaaaga actggggtcc 3901 caaatcctta aggaacaccc agttgaaaac acccagcttc agaatgagaa gctctacctg 3961 tactacctgc agaacggcag ggacatgtac gtggatcagg aactggacat caatcggctc 4021 tccgactacg acgtggatgc tatcgtgccc cagtcttttc tcaaagatga ttctattgat 4081 aataaagtgt tgacaagatc cgataaaaat agagggaaga gtgataacgt cccctcagaa 4141 gaagttgtca agaaaatgaa aaattattgg cggcagctgc tgaacgccaa actgatcaca 4201 caacggaagt tcgataatct gactaaggct gaacgaggtg gcctgtctga gttggataaa 4261 gccggcttca tcaaaaggca gcttgttgag acacgccaga tcaccaagca cgtggcccaa 4321 attctcgatt cacgcatgaa caccaagtac gatgaaaatg acaaactgat tcgagaggtg 4381 aaagttatta ctctgaagtc taagctggtc tcagatttca gaaaggactt tcagttttat 4441 aaggtgagag agatcaacaa ttaccaccat gcgcatgatg cctacctgaa tgcagtggta 4501 ggcactgcac ttatcaaaaa atatcccaag cttgaatctg aatttgttta cggagactat 4561 aaagtgtacg atgttaggaa aatgatcgca aagtctgagc aggaaatagg caaggccacc 4621 gctaagtact tcttttacag caatattatg aattttttca agaccgagat tacactggcc 4681 aatggagaga ttcggaagcg accacttatc gaaacaaacg gagaaacagg agaaatcgtg 4741 tgggacaagg gtagggattt cgcgacagtc cggaaggtcc tgtccatgcc gcaggtgaac 4801 atcgttaaaa agaccgaagt acagaccgga ggcttctcca aggaaagtat cctcccgaaa 4861 aggaacagcg acaagctgat cgcacgcaaa aaagattggg accccaagaa atacggcgga 4921 ttcgattctc ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga gaaagggaag 4981 tctaaaaaac tcaaaagcgt caaggaactg ctgggcatca caatcatgga gcgatcaagc 5041 ttcgaaaaaa accccatcga ctttctcgag gcgaaaggat ataaagaggt caaaaaagac 5101 ctcatcatta agcttcccaa gtactctctc tttgagcttg aaaacggccg gaaacgaatg 5161 ctcgctagtg cgggcgagct gcagaaaggt aacgagctgg cactgccctc taaatacgtt 5221 aatttcttgt atctggccag ccactatgaa aagctcaaag ggtctcccga agataatgag 5281 cagaagcagc tgttcgtgga acaacacaaa cactaccttg atgagatcat cgagcaaata 5341 agcgaattct ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt gctttctgct 5401 tacaataagc acagggataa gcccatcagg gagcaggcag aaaacattat ccacttgttt 5461 actctgacca acttgggcgc gcctgcagcc ttcaagtact tcgacaccac catagacaga 5521 aagcggtaca cctctacaaa ggaggtcctg gacgccacac tgattcatca gtcaattacg 5581 gggctctatg aaacaagaat cgacctctct cagctcggtg gagacggtgg tagtggaggt 5641 tcaggaggat ccggggggag cggagggagc gctagcggtg gaggctccgg aggtggatct 5701 aaaaggccgg cggccacgaa aaaggccggt caggcaaaaa agaaaaaggg tggtagtgga 5761 agcggagcta ctaacttcag cctgctgaag caggctggag acgtggagga gaaccctgga 5821 cctgcggccg catgaaaggg ttcgatccct accggttagt aatgagttta aacgggggag 5881 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa 5941 aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc 6001 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt 6061 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg 6121 tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc 6181 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 6241 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 6301 cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta 6361 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc 6421 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 6481 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat 6541 aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta 6601 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc 6661 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc 6721 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat 6781 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc 6841 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga 6901 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc 6961 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat 7021 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg 7081 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc 7141 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca 7201 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct 7261 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga 7321 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg 7381 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat 7441 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga 7501 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg 7561 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg 7621 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat 7681 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct 7741 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga 7801 cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc 7861 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg 7921 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg 7981 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 8041 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 8101 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt 8161 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 8221 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 8281 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 8341 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 8401 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 8461 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 8521 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 8581 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 8641 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 8701 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct 8761 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 8821 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 8881 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 8941 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 9001 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 9061 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 9121 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 9181 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 9241 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 9301 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 9361 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 9421 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 9481 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 9541 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 9601 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 9661 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 9721 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 9781 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 9841 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 9901 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 9961 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 10021 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 10081 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 10141 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 10201 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 10261 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg 10321 gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc 10381 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag 10441 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag 10501 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt 10561 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga 10621 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg 10681 cccattgacg tcaataatga cgtatgttcc catagtaacg cca //