LOCUS       Exported               12163 bp ds-DNA     circular SYN 20-SEP-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Ezh2[FL]-iCas9v2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12163)
  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..12163
                     /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    667..2901
                     /label=mEZH2
     misc_feature    2962..7065
                     /label=dCas9_D10A_H840A
     misc_feature    7141..7188
                     /label=Nucleoplasmin NLS
                     /note="note=""direct strand 
                     translation""translation=""KRPAATKKAGQAKKKK"""
     misc_feature    7207..7263
                     /label=P2A
     misc_feature    8243..8320
                     /label=SV40 ORI
     CDS             8462..9253
                     /label=Kan/neoR
     misc_feature    complement(9453..9476)
                     /label=pGK-R
     misc_binding    complement(9621..9643)
                     /label=LacO
     promoter        complement(9648..9677)
                     /label=lac
     rep_origin      complement(9983..10611)
                     /direction=LEFT
                     /label=ColE1 origin
     CDS             complement(10763..11422)
                     /label=AmpR
     misc_feature    12071..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 accatgggcc agactgggaa gaaatctgag aagggaccgg tttgttggcg gaagcgtgta
      721 aaatcagagt acatgagact gagacagctc aagaggttca gaagagctga tgaagtaaag
      781 actatgttta gttccaatcg tcagaaaatt ttggaaagaa ctgaaacctt aaaccaagag
      841 tggaagcagc ggaggataca gcctgtgcac atcatgactt ctgtgagctc attgcgcggg
      901 actagggagt gttcagtcac cagtgacttg gattttccag cacaagtcat cccgttaaag
      961 accctgaatg cagtcgcctc ggtgcctata atgtactctt ggtcgccctt acaacagaat
     1021 tttatggtgg aagacgaaac tgttttacat aacattcctt atatggggga tgaagttctg
     1081 gatcaggatg gcactttcat tgaagaacta ataaaaaatt atgatggaaa agtgcatggt
     1141 gacagagaat gtggatttat aaatgatgaa atttttgtgg agttggtaaa tgctcttggt
     1201 caatataatg atgatgatga tgacgatgat ggagatgatc cagatgaaag agaagaaaaa
     1261 cagaaagatc tagaggataa tcgagatgat aaagaaactt gcccacctcg gaaatttcct
     1321 gctgataaaa tatttgaagc catttcctca atgtttccag ataagggcac cgcagaagaa
     1381 ctgaaagaaa aatataaaga actcacggag cagcagctcc caggtgctct gcctcctgaa
     1441 tgtactccaa acatcgatgg accaaatgcc aaatctgttc agagggagca aagcttgcat
     1501 tcatttcata cgctcttctg tcgacgatgt tttaagtatg actgcttcct acatcccttc
     1561 catgcaacac ccaacacata taagaggaag aacacagaaa cagctttgga caacaagcct
     1621 tgtggaccac agtgttacca gcatctggag ggagctaagg agtttgctgc tgctcttact
     1681 gctgagcgta taaagacacc acctaaacgc ccagggggcc gcagaagagg aagacttccg
     1741 aataacagta gcagacccag cacccccacc atcagtgtgc tggagtcaaa ggatacagac
     1801 agtgacagag aagcagggac tgaaactggg ggagagaaca atgataaaga agaagaagag
     1861 aaaaaagatg agacgtccag ctcctctgaa gcaaattctc ggtgtcaaac accaataaag
     1921 atgaagccaa atattgaacc tcctgagaat gtggagtgga gtggtgctga agcctccatg
     1981 tttagagtcc tcattggtac ttactacgat aacttttgtg ccattgctag gctaattggg
     2041 accaaaacat gtagacaggt gtatgagttt agagtcaagg agtccagtat catagcacct
     2101 gttcccactg aggatgtaga cactcctcca agaaagaaga aaaggaaaca tcggttgtgg
     2161 gctgcacact gcagaaagat acaactgaaa aaggacggct cctctaacca tgtttacaac
     2221 tatcaaccct gtgaccatcc acggcagcct tgtgacagtt cgtgcccttg tgtgatagca
     2281 caaaattttt gtgaaaagtt ttgtcaatgt agttcagagt gtcaaaaccg ctttcctgga
     2341 tgtcggtgca aagcacaatg caacaccaaa cagtgtccat gctacctggc tgtccgagag
     2401 tgtgaccctg acctctgtct cacgtgtgga gctgctgacc attgggacag taaaaatgta
     2461 tcctgtaaga actgtagcat tcagcggggc tctaaaaagc acttactgct ggcaccgtct
     2521 gatgtggcag gctggggcat ctttatcaaa gatcctgtac agaaaaatga attcatctca
     2581 gaatactgtg gggagattat ttctcaggat gaagcagaca gaagaggaaa agtgtatgac
     2641 aaatacatgt gcagctttct gttcaacttg aacaatgatt ttgtggtgga tgcaacccga
     2701 aagggcaaca aaattcgttt tgctaatcat tcagtaaatc caaactgcta tgcaaaagtt
     2761 atgatggtta atggtgacca caggataggc atctttgcta agagggctat ccagactggt
     2821 gaagagttgt tttttgatta cagatacagc caggctgatg ccctgaagta tgtgggcatc
     2881 gaacgagaaa tggaaatccc tagtaccgga gggagtggtg gaagcggcgg ttctggtggc
     2941 tcaggcggca gcggccggcc aatggacaag aagtactcca ttgggctcgc tatcggcaca
     3001 aacagcgtcg gctgggccgt cattacggac gagtacaagg tgccgagcaa aaaattcaaa
     3061 gttctgggca ataccgatcg ccacagcata aagaagaacc tcattggcgc cctcctgttc
     3121 gactccgggg agacggccga agccacgcgg ctcaaaagaa cagcacggcg cagatatacc
     3181 cgcagaaaga atcggatctg ctacctgcag gagatcttta gtaatgagat ggctaaggtg
     3241 gatgactctt tcttccatag gctggaggag tcctttttgg tggaggagga taaaaagcac
     3301 gagcgccacc caatctttgg caatatcgtg gacgaggtgg cgtaccatga aaagtaccca
     3361 accatatatc atctgaggaa gaagcttgta gacagtactg ataaggctga cttgcggttg
     3421 atctatctcg cgctggcgca tatgatcaaa tttcggggac acttcctcat cgagggggac
     3481 ctgaacccag acaacagcga tgtcgacaaa ctctttatcc aactggttca gacttacaat
     3541 cagcttttcg aagagaaccc gatcaacgca tccggagttg acgccaaagc aatcctgagc
     3601 gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg cacagctccc tggggagaag
     3661 aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc caactttaaa
     3721 tctaacttcg acctggccga agatgccaag cttcaactga gcaaagacac ctacgatgat
     3781 gatctcgaca atctgctggc ccagatcggc gaccagtacg cagacctttt tttggcggca
     3841 aagaacctgt cagacgccat tctgctgagt gatattctgc gagtgaacac ggagatcacc
     3901 aaagctccgc tgagcgctag tatgatcaag cgctatgatg agcaccacca agacttgact
     3961 ttgctgaagg cccttgtcag acagcaactg cctgagaagt acaaggaaat tttcttcgat
     4021 cagtctaaaa atggctacgc cggatacatt gacggcggag caagccagga ggaattttac
     4081 aaatttatta agcccatctt ggaaaaaatg gacggcaccg aggagctgct ggtaaagctt
     4141 aacagagaag atctgttgcg caaacagcgc actttcgaca atggaagcat cccccaccag
     4201 attcacctgg gcgaactgca cgctatcctc aggcggcaag aggatttcta cccctttttg
     4261 aaagataaca gggaaaagat tgagaaaatc ctcacatttc ggatacccta ctatgtaggc
     4321 cccctcgccc ggggaaattc cagattcgcg tggatgactc gcaaatcaga agagaccatc
     4381 actccctgga acttcgagga agtcgtggat aagggggcct ctgcccagtc cttcatcgaa
     4441 aggatgacta actttgataa aaatctgcct aacgaaaagg tgcttcctaa acactctctg
     4501 ctgtacgagt acttcacagt ttataacgag ctcaccaagg tcaaatacgt cacagaaggg
     4561 atgagaaagc cagcattcct gtctggagag cagaagaaag ctatcgtgga cctcctcttc
     4621 aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag actatttcaa aaagattgaa
     4681 tgtttcgact ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc cctgggaacg
     4741 tatcacgatc tcctgaaaat cattaaagac aaggacttcc tggacaatga ggagaacgag
     4801 gacattcttg aggacattgt cctcaccctt acgttgtttg aagataggga gatgattgaa
     4861 gaacgcttga aaacttacgc tcatctcttc gacgacaaag tcatgaaaca gctcaagagg
     4921 cgccgatata caggatgggg gcggctgtca agaaaactga tcaatgggat ccgagacaag
     4981 cagagtggaa agacaatcct ggattttctt aagtccgatg gatttgccaa ccggaacttc
     5041 atgcagttga tccatgatga ctctctcacc tttaaggagg acatccagaa agcacaagtt
     5101 tctggccagg gggacagtct tcacgagcac atcgctaatc ttgcaggtag cccagctatc
     5161 aaaaagggaa tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt aatgggaagg
     5221 cataagcccg agaatatcgt tatcgagatg gcccgagaga accaaactac ccagaaggga
     5281 cagaagaaca gtagggaaag gatgaagagg attgaagagg gtataaaaga actggggtcc
     5341 caaatcctta aggaacaccc agttgaaaac acccagcttc agaatgagaa gctctacctg
     5401 tactacctgc agaacggcag ggacatgtac gtggatcagg aactggacat caatcggctc
     5461 tccgactacg acgtggatgc tatcgtgccc cagtcttttc tcaaagatga ttctattgat
     5521 aataaagtgt tgacaagatc cgataaaaat agagggaaga gtgataacgt cccctcagaa
     5581 gaagttgtca agaaaatgaa aaattattgg cggcagctgc tgaacgccaa actgatcaca
     5641 caacggaagt tcgataatct gactaaggct gaacgaggtg gcctgtctga gttggataaa
     5701 gccggcttca tcaaaaggca gcttgttgag acacgccaga tcaccaagca cgtggcccaa
     5761 attctcgatt cacgcatgaa caccaagtac gatgaaaatg acaaactgat tcgagaggtg
     5821 aaagttatta ctctgaagtc taagctggtc tcagatttca gaaaggactt tcagttttat
     5881 aaggtgagag agatcaacaa ttaccaccat gcgcatgatg cctacctgaa tgcagtggta
     5941 ggcactgcac ttatcaaaaa atatcccaag cttgaatctg aatttgttta cggagactat
     6001 aaagtgtacg atgttaggaa aatgatcgca aagtctgagc aggaaatagg caaggccacc
     6061 gctaagtact tcttttacag caatattatg aattttttca agaccgagat tacactggcc
     6121 aatggagaga ttcggaagcg accacttatc gaaacaaacg gagaaacagg agaaatcgtg
     6181 tgggacaagg gtagggattt cgcgacagtc cggaaggtcc tgtccatgcc gcaggtgaac
     6241 atcgttaaaa agaccgaagt acagaccgga ggcttctcca aggaaagtat cctcccgaaa
     6301 aggaacagcg acaagctgat cgcacgcaaa aaagattggg accccaagaa atacggcgga
     6361 ttcgattctc ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga gaaagggaag
     6421 tctaaaaaac tcaaaagcgt caaggaactg ctgggcatca caatcatgga gcgatcaagc
     6481 ttcgaaaaaa accccatcga ctttctcgag gcgaaaggat ataaagaggt caaaaaagac
     6541 ctcatcatta agcttcccaa gtactctctc tttgagcttg aaaacggccg gaaacgaatg
     6601 ctcgctagtg cgggcgagct gcagaaaggt aacgagctgg cactgccctc taaatacgtt
     6661 aatttcttgt atctggccag ccactatgaa aagctcaaag ggtctcccga agataatgag
     6721 cagaagcagc tgttcgtgga acaacacaaa cactaccttg atgagatcat cgagcaaata
     6781 agcgaattct ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt gctttctgct
     6841 tacaataagc acagggataa gcccatcagg gagcaggcag aaaacattat ccacttgttt
     6901 actctgacca acttgggcgc gcctgcagcc ttcaagtact tcgacaccac catagacaga
     6961 aagcggtaca cctctacaaa ggaggtcctg gacgccacac tgattcatca gtcaattacg
     7021 gggctctatg aaacaagaat cgacctctct cagctcggtg gagacggtgg tagtggaggt
     7081 tcaggaggat ccggggggag cggagggagc gctagcggtg gaggctccgg aggtggatct
     7141 aaaaggccgg cggccacgaa aaaggccggt caggcaaaaa agaaaaaggg tggtagtgga
     7201 agcggagcta ctaacttcag cctgctgaag caggctggag acgtggagga gaaccctgga
     7261 cctgcggccg catgaaaggg ttcgatccct accggttagt aatgagttta aacgggggag
     7321 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     7381 aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     7441 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     7501 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     7561 tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc
     7621 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     7681 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     7741 cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta
     7801 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     7861 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     7921 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     7981 aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     8041 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     8101 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     8161 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     8221 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     8281 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     8341 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     8401 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     8461 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     8521 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     8581 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     8641 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     8701 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     8761 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     8821 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     8881 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     8941 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     9001 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     9061 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     9121 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     9181 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     9241 cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc
     9301 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     9361 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     9421 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     9481 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     9541 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     9601 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     9661 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     9721 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     9781 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     9841 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     9901 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     9961 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
    10021 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
    10081 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
    10141 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
    10201 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
    10261 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
    10321 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
    10381 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
    10441 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
    10501 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
    10561 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
    10621 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
    10681 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
    10741 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
    10801 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
    10861 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
    10921 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
    10981 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
    11041 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
    11101 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
    11161 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
    11221 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
    11281 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
    11341 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
    11401 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
    11461 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
    11521 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
    11581 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
    11641 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
    11701 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
    11761 gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc
    11821 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
    11881 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
    11941 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt
    12001 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
    12061 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
    12121 cccattgacg tcaataatga cgtatgttcc catagtaacg cca
//
