LOCUS       Exported               10936 bp ds-DNA     circular SYN 20-SEP-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    SUV[SET]-iCas9v2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10936)
  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..10936
                     /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..1674
                     /label=SuvDel1-76
     misc_feature    1735..5838
                     /label=dCas9_D10A_H840A
     misc_feature    5914..5961
                     /label=Nucleoplasmin NLS
                     /note="note=""direct strand 
                     translation""translation=""KRPAATKKAGQAKKKK"""
     misc_feature    5980..6036
                     /label=P2A
     misc_feature    6087..6358
                     /label=TK polyA signal
     misc_feature    7016..7093
                     /label=SV40 ORI
     CDS             7235..8026
                     /label=Kan/neoR
     misc_binding    complement(8394..8416)
                     /label=LacO
     promoter        complement(8421..8450)
                     /label=lac
     rep_origin      complement(8756..9384)
                     /direction=LEFT
                     /label=ColE1 origin
     CDS             complement(9536..10195)
                     /label=AmpR
     misc_feature    10844..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 accccacggc agaatctcaa gtgtgtgcgt atcctcaagc agttccacaa ggacttagaa
      721 agggagctgc tccggcggca ccaccggtca aagacccccc ggcacctgga cccaagcttg
      781 gccaactacc tggtgcagaa ggccaagcag aggcgggcgc tccgtcgctg ggagcaggag
      841 ctcaatgcca agcgcagcca tctgggacgc atcactgtag agaatgaggt ggacctggac
      901 ggccctccgc gggccttcgt gtacatcaat gagtaccgtg ttggtgaggg catcaccctc
      961 aaccaggtgg ctgtgggctg cgagtgccag gactgtctgt gggcacccac tggaggctgc
     1021 tgcccggggg cgtcactgca caagtttgcc tacaatgacc agggccaggt gcggcttcga
     1081 gccgggctgc ccatctacga gtgcaactcc cgctgccgct gcggctatga ctgcccaaat
     1141 cgtgtggtac agaagggtat ccgatatgac ctctgcatct tccgcacgga tgatgggcgt
     1201 ggctggggcg tccgcaccct ggagaagatt cgcaagaaca gcttcgtcat ggagtacgtg
     1261 ggagagatca ttacctcaga ggaggcagag cggcggggcc agatctacga ccgtcagggc
     1321 gccacctacc tctttgacct ggactacgtg gaggacgtgt acaccgtgga tgccgcctac
     1381 tatggcaaca tctcccactt tgtcaaccac agttgtgacc ccaacctgca ggtgtacaac
     1441 gtcttcatag acaaccttga cgagcggctg ccccgcatcg ctttctttgc cacaagaacc
     1501 atccgggcag gcgaggagct cacctttgat tacaacatgc aagtggaccc cgtggacatg
     1561 gagagcaccc gcatggactc caactttggc ctggctgggc tccctggctc ccctaagaag
     1621 cgggtccgta ttgaatgcaa gtgtgggact gagtcctgcc gcaaatacct cttcagtacc
     1681 ggagggagtg gtggaagcgg cggttctggt ggctcaggcg gcagcggccg gccaatggac
     1741 aagaagtact ccattgggct cgctatcggc acaaacagcg tcggctgggc cgtcattacg
     1801 gacgagtaca aggtgccgag caaaaaattc aaagttctgg gcaataccga tcgccacagc
     1861 ataaagaaga acctcattgg cgccctcctg ttcgactccg gggagacggc cgaagccacg
     1921 cggctcaaaa gaacagcacg gcgcagatat acccgcagaa agaatcggat ctgctacctg
     1981 caggagatct ttagtaatga gatggctaag gtggatgact ctttcttcca taggctggag
     2041 gagtcctttt tggtggagga ggataaaaag cacgagcgcc acccaatctt tggcaatatc
     2101 gtggacgagg tggcgtacca tgaaaagtac ccaaccatat atcatctgag gaagaagctt
     2161 gtagacagta ctgataaggc tgacttgcgg ttgatctatc tcgcgctggc gcatatgatc
     2221 aaatttcggg gacacttcct catcgagggg gacctgaacc cagacaacag cgatgtcgac
     2281 aaactcttta tccaactggt tcagacttac aatcagcttt tcgaagagaa cccgatcaac
     2341 gcatccggag ttgacgccaa agcaatcctg agcgctaggc tgtccaaatc ccggcggctc
     2401 gaaaacctca tcgcacagct ccctggggag aagaagaacg gcctgtttgg taatcttatc
     2461 gccctgtcac tcgggctgac ccccaacttt aaatctaact tcgacctggc cgaagatgcc
     2521 aagcttcaac tgagcaaaga cacctacgat gatgatctcg acaatctgct ggcccagatc
     2581 ggcgaccagt acgcagacct ttttttggcg gcaaagaacc tgtcagacgc cattctgctg
     2641 agtgatattc tgcgagtgaa cacggagatc accaaagctc cgctgagcgc tagtatgatc
     2701 aagcgctatg atgagcacca ccaagacttg actttgctga aggcccttgt cagacagcaa
     2761 ctgcctgaga agtacaagga aattttcttc gatcagtcta aaaatggcta cgccggatac
     2821 attgacggcg gagcaagcca ggaggaattt tacaaattta ttaagcccat cttggaaaaa
     2881 atggacggca ccgaggagct gctggtaaag cttaacagag aagatctgtt gcgcaaacag
     2941 cgcactttcg acaatggaag catcccccac cagattcacc tgggcgaact gcacgctatc
     3001 ctcaggcggc aagaggattt ctaccccttt ttgaaagata acagggaaaa gattgagaaa
     3061 atcctcacat ttcggatacc ctactatgta ggccccctcg cccggggaaa ttccagattc
     3121 gcgtggatga ctcgcaaatc agaagagacc atcactccct ggaacttcga ggaagtcgtg
     3181 gataaggggg cctctgccca gtccttcatc gaaaggatga ctaactttga taaaaatctg
     3241 cctaacgaaa aggtgcttcc taaacactct ctgctgtacg agtacttcac agtttataac
     3301 gagctcacca aggtcaaata cgtcacagaa gggatgagaa agccagcatt cctgtctgga
     3361 gagcagaaga aagctatcgt ggacctcctc ttcaagacga accggaaagt taccgtgaaa
     3421 cagctcaaag aagactattt caaaaagatt gaatgtttcg actctgttga aatcagcgga
     3481 gtggaggatc gcttcaacgc atccctggga acgtatcacg atctcctgaa aatcattaaa
     3541 gacaaggact tcctggacaa tgaggagaac gaggacattc ttgaggacat tgtcctcacc
     3601 cttacgttgt ttgaagatag ggagatgatt gaagaacgct tgaaaactta cgctcatctc
     3661 ttcgacgaca aagtcatgaa acagctcaag aggcgccgat atacaggatg ggggcggctg
     3721 tcaagaaaac tgatcaatgg gatccgagac aagcagagtg gaaagacaat cctggatttt
     3781 cttaagtccg atggatttgc caaccggaac ttcatgcagt tgatccatga tgactctctc
     3841 acctttaagg aggacatcca gaaagcacaa gtttctggcc agggggacag tcttcacgag
     3901 cacatcgcta atcttgcagg tagcccagct atcaaaaagg gaatactgca gaccgttaag
     3961 gtcgtggatg aactcgtcaa agtaatggga aggcataagc ccgagaatat cgttatcgag
     4021 atggcccgag agaaccaaac tacccagaag ggacagaaga acagtaggga aaggatgaag
     4081 aggattgaag agggtataaa agaactgggg tcccaaatcc ttaaggaaca cccagttgaa
     4141 aacacccagc ttcagaatga gaagctctac ctgtactacc tgcagaacgg cagggacatg
     4201 tacgtggatc aggaactgga catcaatcgg ctctccgact acgacgtgga tgctatcgtg
     4261 ccccagtctt ttctcaaaga tgattctatt gataataaag tgttgacaag atccgataaa
     4321 aatagaggga agagtgataa cgtcccctca gaagaagttg tcaagaaaat gaaaaattat
     4381 tggcggcagc tgctgaacgc caaactgatc acacaacgga agttcgataa tctgactaag
     4441 gctgaacgag gtggcctgtc tgagttggat aaagccggct tcatcaaaag gcagcttgtt
     4501 gagacacgcc agatcaccaa gcacgtggcc caaattctcg attcacgcat gaacaccaag
     4561 tacgatgaaa atgacaaact gattcgagag gtgaaagtta ttactctgaa gtctaagctg
     4621 gtctcagatt tcagaaagga ctttcagttt tataaggtga gagagatcaa caattaccac
     4681 catgcgcatg atgcctacct gaatgcagtg gtaggcactg cacttatcaa aaaatatccc
     4741 aagcttgaat ctgaatttgt ttacggagac tataaagtgt acgatgttag gaaaatgatc
     4801 gcaaagtctg agcaggaaat aggcaaggcc accgctaagt acttctttta cagcaatatt
     4861 atgaattttt tcaagaccga gattacactg gccaatggag agattcggaa gcgaccactt
     4921 atcgaaacaa acggagaaac aggagaaatc gtgtgggaca agggtaggga tttcgcgaca
     4981 gtccggaagg tcctgtccat gccgcaggtg aacatcgtta aaaagaccga agtacagacc
     5041 ggaggcttct ccaaggaaag tatcctcccg aaaaggaaca gcgacaagct gatcgcacgc
     5101 aaaaaagatt gggaccccaa gaaatacggc ggattcgatt ctcctacagt cgcttacagt
     5161 gtactggttg tggccaaagt ggagaaaggg aagtctaaaa aactcaaaag cgtcaaggaa
     5221 ctgctgggca tcacaatcat ggagcgatca agcttcgaaa aaaaccccat cgactttctc
     5281 gaggcgaaag gatataaaga ggtcaaaaaa gacctcatca ttaagcttcc caagtactct
     5341 ctctttgagc ttgaaaacgg ccggaaacga atgctcgcta gtgcgggcga gctgcagaaa
     5401 ggtaacgagc tggcactgcc ctctaaatac gttaatttct tgtatctggc cagccactat
     5461 gaaaagctca aagggtctcc cgaagataat gagcagaagc agctgttcgt ggaacaacac
     5521 aaacactacc ttgatgagat catcgagcaa ataagcgaat tctccaaaag agtgatcctc
     5581 gccgacgcta acctcgataa ggtgctttct gcttacaata agcacaggga taagcccatc
     5641 agggagcagg cagaaaacat tatccacttg tttactctga ccaacttggg cgcgcctgca
     5701 gccttcaagt acttcgacac caccatagac agaaagcggt acacctctac aaaggaggtc
     5761 ctggacgcca cactgattca tcagtcaatt acggggctct atgaaacaag aatcgacctc
     5821 tctcagctcg gtggagacgg tggtagtgga ggttcaggag gatccggggg gagcggaggg
     5881 agcgctagcg gtggaggctc cggaggtgga tctaaaaggc cggcggccac gaaaaaggcc
     5941 ggtcaggcaa aaaagaaaaa gggtggtagt ggaagcggag ctactaactt cagcctgctg
     6001 aagcaggctg gagacgtgga ggagaaccct ggacctgcgg ccgcatgaaa gggttcgatc
     6061 cctaccggtt agtaatgagt ttaaacgggg gaggctaact gaaacacgga aggagacaat
     6121 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg
     6181 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     6241 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     6301 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag
     6361 atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     6421 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     6481 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     6541 taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     6601 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     6661 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     6721 tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt
     6781 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     6841 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     6901 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     6961 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     7021 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     7081 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     7141 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     7201 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     7261 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     7321 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     7381 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     7441 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     7501 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     7561 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     7621 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     7681 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     7741 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     7801 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     7861 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     7921 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     7981 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     8041 ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     8101 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     8161 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     8221 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     8281 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     8341 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     8401 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     8461 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     8521 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     8581 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     8641 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     8701 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     8761 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     8821 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     8881 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     8941 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     9001 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     9061 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     9121 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     9181 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     9241 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     9301 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     9361 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     9421 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     9481 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     9541 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     9601 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     9661 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     9721 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     9781 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     9841 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     9901 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     9961 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
    10021 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
    10081 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
    10141 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
    10201 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
    10261 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
    10321 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
    10381 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
    10441 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
    10501 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
    10561 gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
    10621 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
    10681 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
    10741 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
    10801 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
    10861 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
    10921 tcccatagta acgcca
//
