LOCUS       Exported               10762 bp ds-DNA     circular SYN 19-AUG-2020
DEFINITION  pcDNA3.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-dCas9:gs:NLS:10P3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10762)
  AUTHORS   UNKNOWN
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Aug 19, 2020 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10762
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        236..852
                     /label=CMV_immearly_promoter
     misc_feature    315..602
                     /label=CAG_enhancer
     misc_feature    769..789
                     /label=CMV_fwd_primer
     promoter        863..881
                     /label=T7_promoter
     misc_signal     944..949
                     /label=Kozak
     misc_signal     953..970
                     /label=His-tag
     misc_feature    971..5068
                     /label=dCas9
     misc_signal     5072..5095
                     /label=NLS
     misc_feature    5096..5194
                     /label=P3
     misc_feature    5219..5317
                     /label=P3
     misc_feature    5342..5440
                     /label=P3
     misc_feature    5465..5563
                     /label=P3
     misc_feature    5588..5686
                     /label=P3
     misc_feature    5705..5803
                     /label=P3
     misc_feature    5828..5926
                     /label=P3
     misc_feature    5951..6049
                     /label=P3
     misc_feature    6074..6172
                     /label=P3
     misc_feature    6197..6295
                     /label=P3
     misc_feature    complement(6315..6332)
                     /label=Sp6_primer
     promoter        complement(6316..6333)
                     /label=Sp6_promoter
     misc_feature    complement(6352..6369)
                     /label=BGH_rev_primer
     terminator      6355..6582
                     /label=bGH_PA_terminator
     rep_origin      6645..6951
                     /label=f1_origin
     misc_feature    complement(7065..7085)
                     /label=pBABE_3_primer
     misc_feature    complement(7071..7287)
                     /label=SV40_enhancer
     promoter        7083..7352
                     /label=SV40_promoter
     rep_origin      7251..7328
                     /label=SV40_origin
     misc_feature    7313..7332
                     /label=SV40pro_F_primer
     CDS             7467..8261
                     /gene="Neomycin"
                     /label=Neomycin
                     /note="ORF frame 3"
     gene            7470..8258
                     /gene="Neomycin"
                     /label=Neomycin
     terminator      8438..8557
                     /label=SV40_PA_terminator
     misc_feature    8526..8545
                     /label=EBV_rev_primer
     misc_feature    complement(8601..8619)
                     /label=M13_reverse_primer
     misc_feature    complement(8618..8640)
                     /label=M13_pUC_rev_primer
     promoter        complement(8654..8683)
                     /label=lac_promoter
     rep_origin      complement(8992..9611)
                     /direction=LEFT
                     /label=pBR322_origin
     CDS             complement(9766..10626)
                     /gene="Ampicillin"
                     /label=Ampicillin
                     /note="ORF frame 1"
     gene            complement(9766..10626)
                     /gene="Ampicillin"
                     /label=Ampicillin
     promoter        complement(10668..10696)
                     /label=AmpR_promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcgccacca tgcaccacca
      961 ccaccaccac gacaagaagt acagcatcgg cctggccatc ggcaccaact ctgtgggctg
     1021 ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac
     1081 cgaccggcac agcatcaaga agaacctgat cggcgccctg ctgttcgaca gcggagaaac
     1141 agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg
     1201 gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt
     1261 ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat
     1321 cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct
     1381 gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct
     1441 ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa
     1501 cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga
     1561 aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa
     1621 gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt
     1681 cggcaacctg attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct
     1741 ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct
     1801 gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga
     1861 cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag
     1921 cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct
     1981 cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg
     2041 ctacgccggc tacatcgatg gcggagccag ccaggaagag ttctacaagt tcatcaagcc
     2101 catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct
     2161 gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga
     2221 gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga
     2281 aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg
     2341 aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt
     2401 cgaggaagtg gtggacaagg gcgccagcgc ccagagcttc atcgagcgga tgaccaactt
     2461 cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt
     2521 caccgtgtac aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc
     2581 cttcctgagc ggcgagcaga aaaaagccat cgtggacctg ctgttcaaga ccaaccggaa
     2641 agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt
     2701 ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct
     2761 gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga
     2821 tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac
     2881 ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg
     2941 ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac
     3001 aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca
     3061 cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga
     3121 tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct
     3181 gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa
     3241 catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg
     3301 cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga
     3361 acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa
     3421 tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt
     3481 ggacgctatc gtgcctcaga gctttctgaa ggacgactcc atcgataaca aagtgctgac
     3541 tcggagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa
     3601 gatgaagaac tactggcgcc agctgctgaa tgccaagctg attacccaga ggaagttcga
     3661 caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa
     3721 gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg
     3781 gatgaacact aagtacgacg agaacgacaa actgatccgg gaagtgaaag tgatcaccct
     3841 gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat
     3901 caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat
     3961 caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt
     4021 gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt
     4081 ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg
     4141 gaagcggcct ctgatcgaga caaacggcga aacaggcgag atcgtgtggg ataagggccg
     4201 ggactttgcc accgtgcgga aagtgctgtc tatgccccaa gtgaatatcg tgaaaaagac
     4261 cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgacaa
     4321 gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac
     4381 cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa
     4441 gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc
     4501 catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct
     4561 gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg
     4621 cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct
     4681 ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt
     4741 tgtggaacag cacaaacact acctggacga gatcatcgag cagatcagcg agttctccaa
     4801 gagagtgatc ctggccgacg ctaatctgga caaggtgctg agcgcctaca acaagcacag
     4861 agacaagcct atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct
     4921 gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag
     4981 caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac
     5041 acggatcgac ctgtctcagc tgggaggctc tgatccaaaa aagaagagaa aggtatcccc
     5101 ggaagatgag atccagcaac tggaagaaga aatcgctcag ctggaacaga aaaacgcagc
     5161 gctgaaagag aaaaaccagg cgctgaaata cggtcttaag agcggccccg gcagcggctc
     5221 cccggaagat gagatccagc aactggaaga agaaatcgct cagctggaac agaaaaacgc
     5281 agcgctgaaa gagaaaaacc aggcgctgaa atacggtacc ggtagcggcc ccggcagcgg
     5341 ctccccggaa gatgagatcc agcaactgga agaagaaatc gctcagctgg aacagaaaaa
     5401 cgcagcgctg aaagagaaaa accaggcgct gaaatacggt gctagcagcg gccccggcag
     5461 cggctccccg gaagatgaga tccagcaact ggaagaagaa atcgctcagc tggaacagaa
     5521 aaacgcagcg ctgaaagaga aaaaccaggc gctgaaatac ggtctcgaga gcggccccgg
     5581 cagcggctcc ccggaagatg agatccagca actggaagaa gaaatcgctc agctggaaca
     5641 gaaaaacgca gcgctgaaag agaaaaacca ggcgctgaaa tacggtagcg gccccggcag
     5701 cggctccccg gaagatgaga tccagcaact ggaagaagaa atcgctcagc tggaacagaa
     5761 aaacgcagcg ctgaaagaga aaaaccaggc gctgaaatac ggtctcgaga gcggccccgg
     5821 cagcggctcc ccggaagatg agatccagca actggaagaa gaaatcgctc agctggaaca
     5881 gaaaaacgca gcgctgaaag agaaaaacca ggcgctgaaa tacggtgcta gcagcggccc
     5941 cggcagcggc tccccggaag atgagatcca gcaactggaa gaagaaatcg ctcagctgga
     6001 acagaaaaac gcagcgctga aagagaaaaa ccaggcgctg aaatacggta ccggtagcgg
     6061 ccccggcagc ggctccccgg aagatgagat ccagcaactg gaagaagaaa tcgctcagct
     6121 ggaacagaaa aacgcagcgc tgaaagagaa aaaccaggcg ctgaaatacg gtcttaagag
     6181 cggccccggc agcggctccc cggaagatga gatccagcaa ctggaagaag aaatcgctca
     6241 gctggaacag aaaaacgcag cgctgaaaga gaaaaaccag gcgctgaaat acggttaatc
     6301 tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg
     6361 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     6421 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     6481 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     6541 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     6601 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     6661 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     6721 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     6781 ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     6841 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     6901 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     6961 ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa
     7021 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     7081 agggtgtgga aagtccccag gctccccagg caggcagaag tatgcaaagc atgcatctca
     7141 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     7201 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     7261 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     7321 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     7381 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     7441 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     7501 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     7561 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     7621 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     7681 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     7741 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     7801 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     7861 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     7921 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     7981 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     8041 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     8101 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     8161 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     8221 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     8281 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     8341 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     8401 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     8461 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     8521 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     8581 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     8641 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     8701 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     8761 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     8821 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     8881 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     8941 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     9001 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     9061 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     9121 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     9181 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     9241 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     9301 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     9361 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     9421 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     9481 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     9541 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     9601 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     9661 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     9721 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     9781 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     9841 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     9901 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     9961 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
    10021 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
    10081 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
    10141 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
    10201 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
    10261 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
    10321 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
    10381 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
    10441 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
    10501 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
    10561 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
    10621 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
    10681 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
    10741 ccgaaaagtg ccacctgacg tc
//
