LOCUS       Exported               11868 bp DNA     circular SYN 30-JUN-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11868)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 30, 2022 from SnapGene Viewer 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..199
                     /label=CMV promoter
     LTR             217..397
                     /label=5' LTR (truncated)
     misc_feature    444..569
                     /label=HIV-1 Psi
     misc_feature    553..917
                     /label=gag
     misc_feature    1062..1295
                     /label=RRE
     misc_feature    1822..1939
                     /label=cPPT/CTS
     promoter        2197..2408
                     /label=EF-1-alpha core promoter
     regulatory      2427..2435
                     /label=vertebrate consensus sequence for strong initia
                     /label=vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)
     CDS             2436..5591
                     /label=SaCas9
     CDS             5592..5639
                     /label=nucleoplasmin NLS
     CDS             5640..5663
                     /label=FLAG
     CDS             5673..5729
                     /label=P2A
     CDS             5730..6326
                     /label=PuroR
     misc_feature    6342..6930
                     /label=WPRE
     LTR             7002..7235
                     /label=3' LTR (Delta-U3)
     polyA_signal    7267..7491
                     /label=bGH poly(A) signal
     rep_origin      7537..7965
                     /label=f1 ori
     promoter        7979..8308
                     /label=SV40 promoter
     rep_origin      8159..8294
                     /label=SV40 ori
     promoter        8356..8403
                     /label=EM7 promoter
     CDS             8422..8796
                     /label=BleoR
     polyA_signal    8926..9047
                     /label=SV40 poly(A) signal
     protein_bind    9120..9136
                     /label=lac operator
     promoter        complement(9144..9174)
                     /label=lac promoter
     protein_bind    9189..9210
                     /label=CAP binding site
     rep_origin      complement(9498..10086)
                     /direction=LEFT
                     /label=ori
     rep_origin      9685..9910
                     /label=ColE1
     CDS             complement(10257..11117)
                     /label=AmpR
     promoter        complement(11118..11222)
                     /label=AmpR promoter
     enhancer        11488..11867
                     /label=CMV enhancer
ORIGIN
        1 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
       61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      181 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
      241 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
      301 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
      361 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
      421 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
      481 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
      541 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
      601 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
      661 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
      721 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
      781 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
      841 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
      901 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
      961 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1021 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1081 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1141 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1201 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1261 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1321 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1381 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1441 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1501 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     1561 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     1621 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     1681 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     1741 agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca
     1801 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     1861 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     1921 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggttaat
     1981 taaggtacct gttccattaa ctgcgctaaa aataattttt aaatcttttt tagcttcttg
     2041 ctcttttttg tacgtctctg ttttagagct agaaatagca agttaaaata aggctagtcc
     2101 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgaattcgct agctaggtct
     2161 tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
     2221 tccccgagaa gttgggggga ggggtcggca attgatccgg tgcctagaga aggtggcgcg
     2281 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
     2341 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
     2401 gaacacagga ccggttctag agcgctgcca ccatgaaaag gaactacatt ctggggctgg
     2461 acatcgggat tacaagcgtg gggtatggga ttattgacta tgaaacaagg gacgtgatcg
     2521 acgcaggcgt cagactgttc aaggaggcca acgtggaaaa caatgaggga cggagaagca
     2581 agaggggagc caggcgcctg aaacgacgga gaaggcacag aatccagagg gtgaagaaac
     2641 tgctgttcga ttacaacctg ctgaccgacc attctgagct gagtggaatt aatccttatg
     2701 aagccagggt gaaaggcctg agtcagaagc tgtcagagga agagttttcc gcagctctgc
     2761 tgcacctggc taagcgccga ggagtgcata acgtcaatga ggtggaagag gacaccggca
     2821 acgagctgtc tacaaaggaa cagatctcac gcaatagcaa agctctggaa gagaagtatg
     2881 tcgcagagct gcagctggaa cggctgaaga aagatggcga ggtgagaggg tcaattaata
     2941 ggttcaagac aagcgactac gtcaaagaag ccaagcagct gctgaaagtg cagaaggctt
     3001 accaccagct ggatcagagc ttcatcgata cttatatcga cctgctggag actcggagaa
     3061 cctactatga gggaccagga gaagggagcc ccttcggatg gaaagacatc aaggaatggt
     3121 acgagatgct gatgggacat tgcacctatt ttccagaaga gctgagaagc gtcaagtacg
     3181 cttataacgc agatctgtac aacgccctga atgacctgaa caacctggtc atcaccaggg
     3241 atgaaaacga gaaactggaa tactatgaga agttccagat catcgaaaac gtgtttaagc
     3301 agaagaaaaa gcctacactg aaacagattg ctaaggagat cctggtcaac gaagaggaca
     3361 tcaagggcta ccgggtgaca agcactggaa aaccagagtt caccaatctg aaagtgtatc
     3421 acgatattaa ggacatcaca gcacggaaag aaatcattga gaacgccgaa ctgctggatc
     3481 agattgctaa gatcctgact atctaccaga gctccgagga catccaggaa gagctgacta
     3541 acctgaacag cgagctgacc caggaagaga tcgaacagat tagtaatctg aaggggtaca
     3601 ccggaacaca caacctgtcc ctgaaagcta tcaatctgat tctggatgag ctgtggcata
     3661 caaacgacaa tcagattgca atctttaacc ggctgaagct ggtcccaaaa aaggtggacc
     3721 tgagtcagca gaaagagatc ccaaccacac tggtggacga tttcattctg tcacccgtgg
     3781 tcaagcggag cttcatccag agcatcaaag tgatcaacgc catcatcaag aagtacggcc
     3841 tgcccaatga tatcattatc gagctggcta gggagaagaa cagcaaggac gcacagaaga
     3901 tgatcaatga gatgcagaaa cgaaaccggc agaccaatga acgcattgaa gagattatcc
     3961 gaactaccgg gaaagagaac gcaaagtacc tgattgaaaa aatcaagctg cacgatatgc
     4021 aggagggaaa gtgtctgtat tctctggagg ccatccccct ggaggacctg ctgaacaatc
     4081 cattcaacta cgaggtcgat catattatcc ccagaagcgt gtccttcgac aattccttta
     4141 acaacaaggt gctggtcaag caggaagaga actctaaaaa gggcaatagg actcctttcc
     4201 agtacctgtc tagttcagat tccaagatct cttacgaaac ctttaaaaag cacattctga
     4261 atctggccaa aggaaagggc cgcatcagca agaccaaaaa ggagtacctg ctggaagagc
     4321 gggacatcaa cagattctcc gtccagaagg attttattaa ccggaatctg gtggacacaa
     4381 gatacgctac tcgcggcctg atgaatctgc tgcgatccta tttccgggtg aacaatctgg
     4441 atgtgaaagt caagtccatc aacggcgggt tcacatcttt tctgaggcgc aaatggaagt
     4501 ttaaaaagga gcgcaacaaa gggtacaagc accatgccga agatgctctg attatcgcaa
     4561 atgccgactt catctttaag gagtggaaaa agctggacaa agccaagaaa gtgatggaga
     4621 accagatgtt cgaagagaag caggccgaat ctatgcccga aatcgagaca gaacaggagt
     4681 acaaggagat tttcatcact cctcaccaga tcaagcatat caaggatttc aaggactaca
     4741 agtactctca ccgggtggat aaaaagccca acagagagct gatcaatgac accctgtata
     4801 gtacaagaaa agacgataag gggaataccc tgattgtgaa caatctgaac ggactgtacg
     4861 acaaagataa tgacaagctg aaaaagctga tcaacaaaag tcccgagaag ctgctgatgt
     4921 accaccatga tcctcagaca tatcagaaac tgaagctgat tatggagcag tacggcgacg
     4981 agaagaaccc actgtataag tactatgaag agactgggaa ctacctgacc aagtatagca
     5041 aaaaggataa tggccccgtg atcaagaaga tcaagtacta tgggaacaag ctgaatgccc
     5101 atctggacat cacagacgat taccctaaca gtcgcaacaa ggtggtcaag ctgtcactga
     5161 agccatacag attcgatgtc tatctggaca acggcgtgta taaatttgtg actgtcaaga
     5221 atctggatgt catcaaaaag gagaactact atgaagtgaa tagcaagtgc tacgaagagg
     5281 ctaaaaagct gaaaaagatt agcaaccagg cagagttcat cgcctccttt tacaacaacg
     5341 acctgattaa gatcaatggc gaactgtata gggtcatcgg ggtgaacaat gatctgctga
     5401 accgcattga agtgaatatg attgacatca cttaccgaga gtatctggaa aacatgaatg
     5461 ataagcgccc ccctcgaatt atcaaaacaa ttgcctctaa gactcagagt atcaaaaagt
     5521 actcaaccga cattctggga aacctgtatg aggtgaagag caaaaagcac cctcagatta
     5581 tcaaaaaggg caagcgacct gccgccacaa agaaggctgg acaggctaag aagaagaaag
     5641 attacaaaga cgatgacgat aagggatccg gcgcaacaaa cttctctctg ctgaaacaag
     5701 ccggagatgt cgaagagaat cctggaccga ccgagtacaa gcccacggtg cgcctcgcca
     5761 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     5821 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     5881 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     5941 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     6001 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     6061 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggagtctcgc
     6121 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     6181 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     6241 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     6301 gcatgacccg caagcccggt gcctgaacgc gttaagtcga caatcaacct ctggattaca
     6361 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     6421 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     6481 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     6541 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     6601 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     6661 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     6721 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     6781 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     6841 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     6901 gtcggatctc cctttgggcc gcctccccgc gtcgacttta agaccaatga cttacaaggc
     6961 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     7021 ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
     7081 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     7141 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     7201 cagacccttt tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag
     7261 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     7321 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     7381 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     7441 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     7501 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     7561 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     7621 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     7681 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     7741 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     7801 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     7861 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     7921 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     7981 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     8041 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     8101 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     8161 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     8221 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     8281 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     8341 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac
     8401 gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg
     8461 cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc
     8521 gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc
     8581 caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag
     8641 ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc
     8701 atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc
     8761 aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc
     8821 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     8881 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     8941 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     9001 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     9061 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     9121 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     9181 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     9241 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     9301 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     9361 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     9421 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     9481 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     9541 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     9601 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     9661 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     9721 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     9781 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     9841 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     9901 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     9961 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
    10021 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
    10081 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
    10141 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
    10201 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
    10261 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
    10321 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
    10381 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
    10441 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
    10501 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
    10561 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
    10621 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
    10681 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
    10741 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
    10801 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
    10861 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
    10921 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
    10981 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
    11041 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
    11101 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
    11161 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
    11221 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
    11281 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
    11341 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
    11401 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
    11461 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
    11521 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
    11581 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
    11641 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
    11701 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
    11761 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
    11821 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgg
//
