LOCUS       Exported                9919 bp ds-DNA     circular SYN 20-SEP-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    iCas9v2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9919)
  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..9919
                     /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    718..4821
                     /label=dCas9_D10A_H840A
     misc_feature    4897..4944
                     /label=Nucleoplasmin NLS
                     /note="note=""direct strand 
                     translation""translation=""KRPAATKKAGQAKKKK"""
     misc_feature    4963..5019
                     /label=P2A
     misc_feature    5999..6076
                     /label=SV40 ORI
     CDS             6218..7009
                     /label=Kan/neoR
     misc_binding    complement(7377..7399)
                     /label=LacO
     promoter        complement(7404..7433)
                     /label=lac
     rep_origin      complement(7739..8367)
                     /direction=LEFT
                     /label=ColE1 origin
     CDS             complement(8519..9178)
                     /label=AmpR
     misc_feature    9827..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 accggaggga gtggtggaag cggcggttct ggtggctcag gcggcagcgg ccggccaatg
      721 gacaagaagt actccattgg gctcgctatc ggcacaaaca gcgtcggctg ggccgtcatt
      781 acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc tgggcaatac cgatcgccac
      841 agcataaaga agaacctcat tggcgccctc ctgttcgact ccggggagac ggccgaagcc
      901 acgcggctca aaagaacagc acggcgcaga tatacccgca gaaagaatcg gatctgctac
      961 ctgcaggaga tctttagtaa tgagatggct aaggtggatg actctttctt ccataggctg
     1021 gaggagtcct ttttggtgga ggaggataaa aagcacgagc gccacccaat ctttggcaat
     1081 atcgtggacg aggtggcgta ccatgaaaag tacccaacca tatatcatct gaggaagaag
     1141 cttgtagaca gtactgataa ggctgacttg cggttgatct atctcgcgct ggcgcatatg
     1201 atcaaatttc ggggacactt cctcatcgag ggggacctga acccagacaa cagcgatgtc
     1261 gacaaactct ttatccaact ggttcagact tacaatcagc ttttcgaaga gaacccgatc
     1321 aacgcatccg gagttgacgc caaagcaatc ctgagcgcta ggctgtccaa atcccggcgg
     1381 ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga acggcctgtt tggtaatctt
     1441 atcgccctgt cactcgggct gacccccaac tttaaatcta acttcgacct ggccgaagat
     1501 gccaagcttc aactgagcaa agacacctac gatgatgatc tcgacaatct gctggcccag
     1561 atcggcgacc agtacgcaga cctttttttg gcggcaaaga acctgtcaga cgccattctg
     1621 ctgagtgata ttctgcgagt gaacacggag atcaccaaag ctccgctgag cgctagtatg
     1681 atcaagcgct atgatgagca ccaccaagac ttgactttgc tgaaggccct tgtcagacag
     1741 caactgcctg agaagtacaa ggaaattttc ttcgatcagt ctaaaaatgg ctacgccgga
     1801 tacattgacg gcggagcaag ccaggaggaa ttttacaaat ttattaagcc catcttggaa
     1861 aaaatggacg gcaccgagga gctgctggta aagcttaaca gagaagatct gttgcgcaaa
     1921 cagcgcactt tcgacaatgg aagcatcccc caccagattc acctgggcga actgcacgct
     1981 atcctcaggc ggcaagagga tttctacccc tttttgaaag ataacaggga aaagattgag
     2041 aaaatcctca catttcggat accctactat gtaggccccc tcgcccgggg aaattccaga
     2101 ttcgcgtgga tgactcgcaa atcagaagag accatcactc cctggaactt cgaggaagtc
     2161 gtggataagg gggcctctgc ccagtccttc atcgaaagga tgactaactt tgataaaaat
     2221 ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt acgagtactt cacagtttat
     2281 aacgagctca ccaaggtcaa atacgtcaca gaagggatga gaaagccagc attcctgtct
     2341 ggagagcaga agaaagctat cgtggacctc ctcttcaaga cgaaccggaa agttaccgtg
     2401 aaacagctca aagaagacta tttcaaaaag attgaatgtt tcgactctgt tgaaatcagc
     2461 ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc acgatctcct gaaaatcatt
     2521 aaagacaagg acttcctgga caatgaggag aacgaggaca ttcttgagga cattgtcctc
     2581 acccttacgt tgtttgaaga tagggagatg attgaagaac gcttgaaaac ttacgctcat
     2641 ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc gatatacagg atgggggcgg
     2701 ctgtcaagaa aactgatcaa tgggatccga gacaagcaga gtggaaagac aatcctggat
     2761 tttcttaagt ccgatggatt tgccaaccgg aacttcatgc agttgatcca tgatgactct
     2821 ctcaccttta aggaggacat ccagaaagca caagtttctg gccaggggga cagtcttcac
     2881 gagcacatcg ctaatcttgc aggtagccca gctatcaaaa agggaatact gcagaccgtt
     2941 aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata agcccgagaa tatcgttatc
     3001 gagatggccc gagagaacca aactacccag aagggacaga agaacagtag ggaaaggatg
     3061 aagaggattg aagagggtat aaaagaactg gggtcccaaa tccttaagga acacccagtt
     3121 gaaaacaccc agcttcagaa tgagaagctc tacctgtact acctgcagaa cggcagggac
     3181 atgtacgtgg atcaggaact ggacatcaat cggctctccg actacgacgt ggatgctatc
     3241 gtgccccagt cttttctcaa agatgattct attgataata aagtgttgac aagatccgat
     3301 aaaaatagag ggaagagtga taacgtcccc tcagaagaag ttgtcaagaa aatgaaaaat
     3361 tattggcggc agctgctgaa cgccaaactg atcacacaac ggaagttcga taatctgact
     3421 aaggctgaac gaggtggcct gtctgagttg gataaagccg gcttcatcaa aaggcagctt
     3481 gttgagacac gccagatcac caagcacgtg gcccaaattc tcgattcacg catgaacacc
     3541 aagtacgatg aaaatgacaa actgattcga gaggtgaaag ttattactct gaagtctaag
     3601 ctggtctcag atttcagaaa ggactttcag ttttataagg tgagagagat caacaattac
     3661 caccatgcgc atgatgccta cctgaatgca gtggtaggca ctgcacttat caaaaaatat
     3721 cccaagcttg aatctgaatt tgtttacgga gactataaag tgtacgatgt taggaaaatg
     3781 atcgcaaagt ctgagcagga aataggcaag gccaccgcta agtacttctt ttacagcaat
     3841 attatgaatt ttttcaagac cgagattaca ctggccaatg gagagattcg gaagcgacca
     3901 cttatcgaaa caaacggaga aacaggagaa atcgtgtggg acaagggtag ggatttcgcg
     3961 acagtccgga aggtcctgtc catgccgcag gtgaacatcg ttaaaaagac cgaagtacag
     4021 accggaggct tctccaagga aagtatcctc ccgaaaagga acagcgacaa gctgatcgca
     4081 cgcaaaaaag attgggaccc caagaaatac ggcggattcg attctcctac agtcgcttac
     4141 agtgtactgg ttgtggccaa agtggagaaa gggaagtcta aaaaactcaa aagcgtcaag
     4201 gaactgctgg gcatcacaat catggagcga tcaagcttcg aaaaaaaccc catcgacttt
     4261 ctcgaggcga aaggatataa agaggtcaaa aaagacctca tcattaagct tcccaagtac
     4321 tctctctttg agcttgaaaa cggccggaaa cgaatgctcg ctagtgcggg cgagctgcag
     4381 aaaggtaacg agctggcact gccctctaaa tacgttaatt tcttgtatct ggccagccac
     4441 tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga agcagctgtt cgtggaacaa
     4501 cacaaacact accttgatga gatcatcgag caaataagcg aattctccaa aagagtgatc
     4561 ctcgccgacg ctaacctcga taaggtgctt tctgcttaca ataagcacag ggataagccc
     4621 atcagggagc aggcagaaaa cattatccac ttgtttactc tgaccaactt gggcgcgcct
     4681 gcagccttca agtacttcga caccaccata gacagaaagc ggtacacctc tacaaaggag
     4741 gtcctggacg ccacactgat tcatcagtca attacggggc tctatgaaac aagaatcgac
     4801 ctctctcagc tcggtggaga cggtggtagt ggaggttcag gaggatccgg ggggagcgga
     4861 gggagcgcta gcggtggagg ctccggaggt ggatctaaaa ggccggcggc cacgaaaaag
     4921 gccggtcagg caaaaaagaa aaagggtggt agtggaagcg gagctactaa cttcagcctg
     4981 ctgaagcagg ctggagacgt ggaggagaac cctggacctg cggccgcatg aaagggttcg
     5041 atccctaccg gttagtaatg agtttaaacg ggggaggcta actgaaacac ggaaggagac
     5101 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg
     5161 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     5221 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     5281 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     5341 cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     5401 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     5461 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5521 ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5581 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5641 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5701 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct
     5761 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     5821 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     5881 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     5941 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     6001 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     6061 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     6121 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     6181 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     6241 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     6301 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     6361 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     6421 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     6481 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     6541 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     6601 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     6661 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     6721 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     6781 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     6841 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     6901 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     6961 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     7021 gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     7081 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     7141 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     7201 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     7261 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     7321 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     7381 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     7441 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     7501 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     7561 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     7621 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     7681 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     7741 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     7801 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7861 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7921 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     7981 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     8041 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     8101 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     8161 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     8221 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     8281 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     8341 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     8401 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     8461 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     8521 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     8581 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     8641 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     8701 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     8761 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     8821 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     8881 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     8941 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     9001 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     9061 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     9121 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     9181 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     9241 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     9301 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     9361 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     9421 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     9481 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     9541 atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     9601 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     9661 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     9721 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
     9781 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     9841 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     9901 tgttcccata gtaacgcca
//
