LOCUS       Exported               11068 bp ds-DNA     circular SYN 20-SEP-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    G9A[SET]-iCas9v2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11068)
  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..11068
                     /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..1806
                     /label=G9A-SET
     misc_feature    1867..5970
                     /label=dCas9_D10A_H840A
     misc_feature    6046..6093
                     /label=Nucleoplasmin NLS
                     /note="note=""direct strand 
                     translation""translation=""KRPAATKKAGQAKKKK"""
     misc_feature    6112..6168
                     /label=P2A
     misc_feature    7148..7225
                     /label=SV40 ORI
     CDS             7367..8158
                     /label=Kan/neoR
     misc_binding    complement(8526..8548)
                     /label=LacO
     promoter        complement(8553..8582)
                     /label=lac
     rep_origin      complement(8888..9516)
                     /direction=LEFT
                     /label=ColE1 origin
     CDS             complement(9668..10327)
                     /label=AmpR
     misc_feature    10976..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 accggcagcg ccgccatcgc cgaagtcctt ctgaatgcgc gctgtgacct ccatgctgtc
      721 aactaccatg gggacacccc cctgcacatc gcagctcggg agagctacca tgactgcgtg
      781 ctgttattcc tgtcacgtgg ggccaaccct gagctgcgga acaaagaggg ggacacagca
      841 tgggacctga ctcccgagcg ctccgacgtg tggtttgcgc ttcaactcaa ccgcaagctc
      901 cgacttgggg tgggaaatcg ggccatccgc acagagaaga tcatctgccg ggacgtggct
      961 cggggctatg agaacgtgcc cattccctgt gtcaacggtg tggatgggga gccctgccct
     1021 gaggattaca agtacatctc agagaactgc gagacgtcca ccatgaacat cgatcgcaac
     1081 atcacccacc tgcagcactg cacgtgtgtg gacgactgct ctagctccaa ctgcctgtgc
     1141 ggccagctca gcatccggtg ctggtatgac aaggatgggc gattgctcca ggaatttaac
     1201 aagattgagc ctccgctgat tttcgagtgt aaccaggcgt gctcatgctg gagaaactgc
     1261 aagaaccggg tcgtacagag tggcatcaag gtgcggctac agctctaccg aacagccaag
     1321 atgggctggg gggtccgcgc cctgcagacc atcccacagg ggaccttcat ctgcgagtat
     1381 gtcggggagc tgatctctga tgctgaggct gatgtgagag aggatgattc ttacctcttc
     1441 gacttagaca acaaggatgg agaggtgtac tgcatagatg cccgttacta tggcaacatc
     1501 agccgcttca tcaaccacct gtgtgacccc aacatcattc ccgtccgggt cttcatgctg
     1561 caccaagacc tgcgatttcc acgcatcgcc ttcttcagtt cccgagacat ccggactggg
     1621 gaggagctag ggtttgacta tggcgaccgc ttctgggaca tcaaaagcaa atatttcacc
     1681 tgccaatgtg gctctgagaa gtgcaagcac tcagccgaag ccattgccct ggagcagagc
     1741 cgtctggccc gcctggaccc acaccctgag ctgctgcccg agctcggctc cctgccccct
     1801 gtcaacagta ccggagggag tggtggaagc ggcggttctg gtggctcagg cggcagcggc
     1861 cggccaatgg acaagaagta ctccattggg ctcgctatcg gcacaaacag cgtcggctgg
     1921 gccgtcatta cggacgagta caaggtgccg agcaaaaaat tcaaagttct gggcaatacc
     1981 gatcgccaca gcataaagaa gaacctcatt ggcgccctcc tgttcgactc cggggagacg
     2041 gccgaagcca cgcggctcaa aagaacagca cggcgcagat atacccgcag aaagaatcgg
     2101 atctgctacc tgcaggagat ctttagtaat gagatggcta aggtggatga ctctttcttc
     2161 cataggctgg aggagtcctt tttggtggag gaggataaaa agcacgagcg ccacccaatc
     2221 tttggcaata tcgtggacga ggtggcgtac catgaaaagt acccaaccat atatcatctg
     2281 aggaagaagc ttgtagacag tactgataag gctgacttgc ggttgatcta tctcgcgctg
     2341 gcgcatatga tcaaatttcg gggacacttc ctcatcgagg gggacctgaa cccagacaac
     2401 agcgatgtcg acaaactctt tatccaactg gttcagactt acaatcagct tttcgaagag
     2461 aacccgatca acgcatccgg agttgacgcc aaagcaatcc tgagcgctag gctgtccaaa
     2521 tcccggcggc tcgaaaacct catcgcacag ctccctgggg agaagaagaa cggcctgttt
     2581 ggtaatctta tcgccctgtc actcgggctg acccccaact ttaaatctaa cttcgacctg
     2641 gccgaagatg ccaagcttca actgagcaaa gacacctacg atgatgatct cgacaatctg
     2701 ctggcccaga tcggcgacca gtacgcagac ctttttttgg cggcaaagaa cctgtcagac
     2761 gccattctgc tgagtgatat tctgcgagtg aacacggaga tcaccaaagc tccgctgagc
     2821 gctagtatga tcaagcgcta tgatgagcac caccaagact tgactttgct gaaggccctt
     2881 gtcagacagc aactgcctga gaagtacaag gaaattttct tcgatcagtc taaaaatggc
     2941 tacgccggat acattgacgg cggagcaagc caggaggaat tttacaaatt tattaagccc
     3001 atcttggaaa aaatggacgg caccgaggag ctgctggtaa agcttaacag agaagatctg
     3061 ttgcgcaaac agcgcacttt cgacaatgga agcatccccc accagattca cctgggcgaa
     3121 ctgcacgcta tcctcaggcg gcaagaggat ttctacccct ttttgaaaga taacagggaa
     3181 aagattgaga aaatcctcac atttcggata ccctactatg taggccccct cgcccgggga
     3241 aattccagat tcgcgtggat gactcgcaaa tcagaagaga ccatcactcc ctggaacttc
     3301 gaggaagtcg tggataaggg ggcctctgcc cagtccttca tcgaaaggat gactaacttt
     3361 gataaaaatc tgcctaacga aaaggtgctt cctaaacact ctctgctgta cgagtacttc
     3421 acagtttata acgagctcac caaggtcaaa tacgtcacag aagggatgag aaagccagca
     3481 ttcctgtctg gagagcagaa gaaagctatc gtggacctcc tcttcaagac gaaccggaaa
     3541 gttaccgtga aacagctcaa agaagactat ttcaaaaaga ttgaatgttt cgactctgtt
     3601 gaaatcagcg gagtggagga tcgcttcaac gcatccctgg gaacgtatca cgatctcctg
     3661 aaaatcatta aagacaagga cttcctggac aatgaggaga acgaggacat tcttgaggac
     3721 attgtcctca cccttacgtt gtttgaagat agggagatga ttgaagaacg cttgaaaact
     3781 tacgctcatc tcttcgacga caaagtcatg aaacagctca agaggcgccg atatacagga
     3841 tgggggcggc tgtcaagaaa actgatcaat gggatccgag acaagcagag tggaaagaca
     3901 atcctggatt ttcttaagtc cgatggattt gccaaccgga acttcatgca gttgatccat
     3961 gatgactctc tcacctttaa ggaggacatc cagaaagcac aagtttctgg ccagggggac
     4021 agtcttcacg agcacatcgc taatcttgca ggtagcccag ctatcaaaaa gggaatactg
     4081 cagaccgtta aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa gcccgagaat
     4141 atcgttatcg agatggcccg agagaaccaa actacccaga agggacagaa gaacagtagg
     4201 gaaaggatga agaggattga agagggtata aaagaactgg ggtcccaaat ccttaaggaa
     4261 cacccagttg aaaacaccca gcttcagaat gagaagctct acctgtacta cctgcagaac
     4321 ggcagggaca tgtacgtgga tcaggaactg gacatcaatc ggctctccga ctacgacgtg
     4381 gatgctatcg tgccccagtc ttttctcaaa gatgattcta ttgataataa agtgttgaca
     4441 agatccgata aaaatagagg gaagagtgat aacgtcccct cagaagaagt tgtcaagaaa
     4501 atgaaaaatt attggcggca gctgctgaac gccaaactga tcacacaacg gaagttcgat
     4561 aatctgacta aggctgaacg aggtggcctg tctgagttgg ataaagccgg cttcatcaaa
     4621 aggcagcttg ttgagacacg ccagatcacc aagcacgtgg cccaaattct cgattcacgc
     4681 atgaacacca agtacgatga aaatgacaaa ctgattcgag aggtgaaagt tattactctg
     4741 aagtctaagc tggtctcaga tttcagaaag gactttcagt tttataaggt gagagagatc
     4801 aacaattacc accatgcgca tgatgcctac ctgaatgcag tggtaggcac tgcacttatc
     4861 aaaaaatatc ccaagcttga atctgaattt gtttacggag actataaagt gtacgatgtt
     4921 aggaaaatga tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa gtacttcttt
     4981 tacagcaata ttatgaattt tttcaagacc gagattacac tggccaatgg agagattcgg
     5041 aagcgaccac ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga caagggtagg
     5101 gatttcgcga cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt taaaaagacc
     5161 gaagtacaga ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa cagcgacaag
     5221 ctgatcgcac gcaaaaaaga ttgggacccc aagaaatacg gcggattcga ttctcctaca
     5281 gtcgcttaca gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa aaaactcaaa
     5341 agcgtcaagg aactgctggg catcacaatc atggagcgat caagcttcga aaaaaacccc
     5401 atcgactttc tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat cattaagctt
     5461 cccaagtact ctctctttga gcttgaaaac ggccggaaac gaatgctcgc tagtgcgggc
     5521 gagctgcaga aaggtaacga gctggcactg ccctctaaat acgttaattt cttgtatctg
     5581 gccagccact atgaaaagct caaagggtct cccgaagata atgagcagaa gcagctgttc
     5641 gtggaacaac acaaacacta ccttgatgag atcatcgagc aaataagcga attctccaaa
     5701 agagtgatcc tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa taagcacagg
     5761 gataagccca tcagggagca ggcagaaaac attatccact tgtttactct gaccaacttg
     5821 ggcgcgcctg cagccttcaa gtacttcgac accaccatag acagaaagcg gtacacctct
     5881 acaaaggagg tcctggacgc cacactgatt catcagtcaa ttacggggct ctatgaaaca
     5941 agaatcgacc tctctcagct cggtggagac ggtggtagtg gaggttcagg aggatccggg
     6001 gggagcggag ggagcgctag cggtggaggc tccggaggtg gatctaaaag gccggcggcc
     6061 acgaaaaagg ccggtcaggc aaaaaagaaa aagggtggta gtggaagcgg agctactaac
     6121 ttcagcctgc tgaagcaggc tggagacgtg gaggagaacc ctggacctgc ggccgcatga
     6181 aagggttcga tccctaccgg ttagtaatga gtttaaacgg gggaggctaa ctgaaacacg
     6241 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     6301 gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     6361 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     6421 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     6481 ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg
     6541 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     6601 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     6661 cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc
     6721 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     6781 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     6841 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgggga
     6901 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     6961 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     7021 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     7081 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     7141 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     7201 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     7261 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     7321 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     7381 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     7441 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     7501 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     7561 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     7621 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     7681 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     7741 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     7801 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     7861 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     7921 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     7981 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     8041 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     8101 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     8161 gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     8221 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     8281 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     8341 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     8401 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     8461 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     8521 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     8581 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     8641 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     8701 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     8761 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     8821 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     8881 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     8941 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     9001 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     9061 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     9121 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     9181 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     9241 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     9301 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     9361 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     9421 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     9481 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     9541 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     9601 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     9661 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     9721 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     9781 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     9841 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     9901 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     9961 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
    10021 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
    10081 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
    10141 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
    10201 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
    10261 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
    10321 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
    10381 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
    10441 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
    10501 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
    10561 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
    10621 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
    10681 cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta
    10741 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
    10801 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
    10861 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
    10921 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
    10981 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
    11041 aatgacgtat gttcccatag taacgcca
//
