LOCUS       Exported               10057 bp DNA     circular SYN 22-SEP-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10057)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 22, 2021 from SnapGene 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10057
                     /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..795
                     /label=VP64
     misc_feature    856..4959
                     /label=dCas9_D10A_H840A
     misc_feature    5005..5010
                     /label=NheI
                     /note="note=""direct strand 
                     translation""translation=""AS"""
     misc_feature    5035..5082
                     /label=Nucleoplasmin NLS
                     /note="note=""direct strand 
                     translation""translation=""KRPAATKKAGQAKKKK"""
     misc_feature    5101..5157
                     /label=P2A
     misc_feature    5958..6173
                     /label=SV40 enhancer-pBabe
     misc_feature    6137..6214
                     /label=SV40 ORI
     CDS             6356..7147
                     /label=Kan/neoR
     primer_bind     complement(7489..7509)
                     /label=M13-rev
     misc_binding    complement(7515..7537)
                     /label=LacO
     promoter        complement(7542..7571)
                     /label=lac
     rep_origin      complement(7877..8505)
                     /direction=LEFT
                     /label=ColE1 origin
     CDS             complement(8657..9316)
                     /label=AmpR
     promoter        complement(9556..9584)
                     /label=Amp prom
     misc_feature    join(9965..10057,1..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 accgacgctt tggatgactt tgatttggac atgctggatg ctctagatga cttcgacctg
      721 gatatgctgg acgcacttga cgactttgac ctcgacatgc tagacgctct ggacgacttc
      781 gatctagaca tgctcagtac cggagggagt ggtggaagcg gcggttctgg tggctcaggc
      841 ggcagcggcc ggccaatgga caagaagtac tccattgggc tcgctatcgg cacaaacagc
      901 gtcggctggg ccgtcattac ggacgagtac aaggtgccga gcaaaaaatt caaagttctg
      961 ggcaataccg atcgccacag cataaagaag aacctcattg gcgccctcct gttcgactcc
     1021 ggggagacgg ccgaagccac gcggctcaaa agaacagcac ggcgcagata tacccgcaga
     1081 aagaatcgga tctgctacct gcaggagatc tttagtaatg agatggctaa ggtggatgac
     1141 tctttcttcc ataggctgga ggagtccttt ttggtggagg aggataaaaa gcacgagcgc
     1201 cacccaatct ttggcaatat cgtggacgag gtggcgtacc atgaaaagta cccaaccata
     1261 tatcatctga ggaagaagct tgtagacagt actgataagg ctgacttgcg gttgatctat
     1321 ctcgcgctgg cgcatatgat caaatttcgg ggacacttcc tcatcgaggg ggacctgaac
     1381 ccagacaaca gcgatgtcga caaactcttt atccaactgg ttcagactta caatcagctt
     1441 ttcgaagaga acccgatcaa cgcatccgga gttgacgcca aagcaatcct gagcgctagg
     1501 ctgtccaaat cccggcggct cgaaaacctc atcgcacagc tccctgggga gaagaagaac
     1561 ggcctgtttg gtaatcttat cgccctgtca ctcgggctga cccccaactt taaatctaac
     1621 ttcgacctgg ccgaagatgc caagcttcaa ctgagcaaag acacctacga tgatgatctc
     1681 gacaatctgc tggcccagat cggcgaccag tacgcagacc tttttttggc ggcaaagaac
     1741 ctgtcagacg ccattctgct gagtgatatt ctgcgagtga acacggagat caccaaagct
     1801 ccgctgagcg ctagtatgat caagcgctat gatgagcacc accaagactt gactttgctg
     1861 aaggcccttg tcagacagca actgcctgag aagtacaagg aaattttctt cgatcagtct
     1921 aaaaatggct acgccggata cattgacggc ggagcaagcc aggaggaatt ttacaaattt
     1981 attaagccca tcttggaaaa aatggacggc accgaggagc tgctggtaaa gcttaacaga
     2041 gaagatctgt tgcgcaaaca gcgcactttc gacaatggaa gcatccccca ccagattcac
     2101 ctgggcgaac tgcacgctat cctcaggcgg caagaggatt tctacccctt tttgaaagat
     2161 aacagggaaa agattgagaa aatcctcaca tttcggatac cctactatgt aggccccctc
     2221 gcccggggaa attccagatt cgcgtggatg actcgcaaat cagaagagac catcactccc
     2281 tggaacttcg aggaagtcgt ggataagggg gcctctgccc agtccttcat cgaaaggatg
     2341 actaactttg ataaaaatct gcctaacgaa aaggtgcttc ctaaacactc tctgctgtac
     2401 gagtacttca cagtttataa cgagctcacc aaggtcaaat acgtcacaga agggatgaga
     2461 aagccagcat tcctgtctgg agagcagaag aaagctatcg tggacctcct cttcaagacg
     2521 aaccggaaag ttaccgtgaa acagctcaaa gaagactatt tcaaaaagat tgaatgtttc
     2581 gactctgttg aaatcagcgg agtggaggat cgcttcaacg catccctggg aacgtatcac
     2641 gatctcctga aaatcattaa agacaaggac ttcctggaca atgaggagaa cgaggacatt
     2701 cttgaggaca ttgtcctcac ccttacgttg tttgaagata gggagatgat tgaagaacgc
     2761 ttgaaaactt acgctcatct cttcgacgac aaagtcatga aacagctcaa gaggcgccga
     2821 tatacaggat gggggcggct gtcaagaaaa ctgatcaatg ggatccgaga caagcagagt
     2881 ggaaagacaa tcctggattt tcttaagtcc gatggatttg ccaaccggaa cttcatgcag
     2941 ttgatccatg atgactctct cacctttaag gaggacatcc agaaagcaca agtttctggc
     3001 cagggggaca gtcttcacga gcacatcgct aatcttgcag gtagcccagc tatcaaaaag
     3061 ggaatactgc agaccgttaa ggtcgtggat gaactcgtca aagtaatggg aaggcataag
     3121 cccgagaata tcgttatcga gatggcccga gagaaccaaa ctacccagaa gggacagaag
     3181 aacagtaggg aaaggatgaa gaggattgaa gagggtataa aagaactggg gtcccaaatc
     3241 cttaaggaac acccagttga aaacacccag cttcagaatg agaagctcta cctgtactac
     3301 ctgcagaacg gcagggacat gtacgtggat caggaactgg acatcaatcg gctctccgac
     3361 tacgacgtgg atgctatcgt gccccagtct tttctcaaag atgattctat tgataataaa
     3421 gtgttgacaa gatccgataa aaatagaggg aagagtgata acgtcccctc agaagaagtt
     3481 gtcaagaaaa tgaaaaatta ttggcggcag ctgctgaacg ccaaactgat cacacaacgg
     3541 aagttcgata atctgactaa ggctgaacga ggtggcctgt ctgagttgga taaagccggc
     3601 ttcatcaaaa ggcagcttgt tgagacacgc cagatcacca agcacgtggc ccaaattctc
     3661 gattcacgca tgaacaccaa gtacgatgaa aatgacaaac tgattcgaga ggtgaaagtt
     3721 attactctga agtctaagct ggtctcagat ttcagaaagg actttcagtt ttataaggtg
     3781 agagagatca acaattacca ccatgcgcat gatgcctacc tgaatgcagt ggtaggcact
     3841 gcacttatca aaaaatatcc caagcttgaa tctgaatttg tttacggaga ctataaagtg
     3901 tacgatgtta ggaaaatgat cgcaaagtct gagcaggaaa taggcaaggc caccgctaag
     3961 tacttctttt acagcaatat tatgaatttt ttcaagaccg agattacact ggccaatgga
     4021 gagattcgga agcgaccact tatcgaaaca aacggagaaa caggagaaat cgtgtgggac
     4081 aagggtaggg atttcgcgac agtccggaag gtcctgtcca tgccgcaggt gaacatcgtt
     4141 aaaaagaccg aagtacagac cggaggcttc tccaaggaaa gtatcctccc gaaaaggaac
     4201 agcgacaagc tgatcgcacg caaaaaagat tgggacccca agaaatacgg cggattcgat
     4261 tctcctacag tcgcttacag tgtactggtt gtggccaaag tggagaaagg gaagtctaaa
     4321 aaactcaaaa gcgtcaagga actgctgggc atcacaatca tggagcgatc aagcttcgaa
     4381 aaaaacccca tcgactttct cgaggcgaaa ggatataaag aggtcaaaaa agacctcatc
     4441 attaagcttc ccaagtactc tctctttgag cttgaaaacg gccggaaacg aatgctcgct
     4501 agtgcgggcg agctgcagaa aggtaacgag ctggcactgc cctctaaata cgttaatttc
     4561 ttgtatctgg ccagccacta tgaaaagctc aaagggtctc ccgaagataa tgagcagaag
     4621 cagctgttcg tggaacaaca caaacactac cttgatgaga tcatcgagca aataagcgaa
     4681 ttctccaaaa gagtgatcct cgccgacgct aacctcgata aggtgctttc tgcttacaat
     4741 aagcacaggg ataagcccat cagggagcag gcagaaaaca ttatccactt gtttactctg
     4801 accaacttgg gcgcgcctgc agccttcaag tacttcgaca ccaccataga cagaaagcgg
     4861 tacacctcta caaaggaggt cctggacgcc acactgattc atcagtcaat tacggggctc
     4921 tatgaaacaa gaatcgacct ctctcagctc ggtggagacg gtggtagtgg aggttcagga
     4981 ggatccgggg ggagcggagg gagcgctagc ggtggaggct ccggaggtgg atctaaaagg
     5041 ccggcggcca cgaaaaaggc cggtcaggca aaaaagaaaa agggtggtag tggaagcgga
     5101 gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctgcg
     5161 gccgcatgaa agggttcgat ccctaccggt tagtaatgag tttaaacggg ggaggctaac
     5221 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     5281 gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     5341 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     5401 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     5461 cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc
     5521 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5581 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5641 ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt
     5701 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     5761 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5821 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     5881 ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5941 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     6001 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     6061 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     6121 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     6181 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     6241 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     6301 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     6361 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     6421 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     6481 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     6541 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     6601 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     6661 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     6721 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     6781 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     6841 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     6901 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     6961 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     7021 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     7081 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     7141 cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat
     7201 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     7261 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     7321 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     7381 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     7441 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     7501 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     7561 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     7621 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     7681 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     7741 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     7801 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     7861 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7921 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7981 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     8041 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     8101 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     8161 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     8221 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     8281 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     8341 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     8401 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     8461 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     8521 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     8581 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     8641 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     8701 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     8761 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     8821 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     8881 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8941 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     9001 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     9061 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     9121 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     9181 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     9241 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     9301 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     9361 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     9421 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     9481 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     9541 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     9601 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     9661 gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt
     9721 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     9781 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     9841 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
     9901 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     9961 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
    10021 gacgtcaata atgacgtatg ttcccatagt aacgcca
//
