LOCUS       pLVX-Puro-Cas9 Vector              12259 bp ds-DNA     circular SYN 01-SEP-2022
DEFINITION  synthetic circular DNA
COMMENT     Created by Takara Bio USA, Inc. http://www.takarabio.com
COMMENT     Cat No. 632631, 632629

FEATURES             Location/Qualifiers
     source          1..12259
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..635
                     /label=5' LTR
     misc_feature    681..806
                     /label=HIV-1 Psi
     misc_feature    1303..1536
                     /label=RRE
     misc_feature    2028..2143
                     /label=cPPT/CTS
     enhancer        2201..2504
                     /label=CMV enhancer
     promoter        2505..2708
                     /label=CMV promoter
     CDS             2896..2916
                     /label=SV40 NLS
     CDS             2941..7041
                     /label=Cas9
     CDS             7054..7074
                     /label=SV40 NLS
     promoter        7094..7593
                     /label=PGK promoter
     CDS             7614..8213
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
     misc_feature    8227..8815
                     /label=WPRE
     LTR             9022..9655
                     /label=3' LTR
     rep_origin      complement(10186..10774)
                     /label=ori
     CDS             complement(10945..11805)
                     /label=AmpR
     promoter        complement(11806..11910)
                     /label=AmpR promoter
     polyA_signal    11958..12092
                     /label=SV40 poly(A) signal
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag
     2821 atggactata aggaccacga cggagactac aaggatcatg atattgatta caaagacgat
     2881 gacgataaga tggccccaaa gaagaagcgg aaggtcggta tccacggagt cccagcagcc
     2941 gacaagaagt actccattgg gctcgatatc ggcacaaaca gcgtcggctg ggccgtcatt
     3001 acggacgact acaaggtgcc gagcaaaaaa ttcaaagttc tgggcaatac cgatcgccac
     3061 agcataaaga agaacctcat tggcgccctc ctgttcgact ccggggagac ggccgaagcc
     3121 acgcggctca aaagaacagc acggcgcaga tatacccgca gaaagaatcg gatctgctac
     3181 ctgcaggaga tctttagtaa tgagatggct aaggtggatg actctttctt ccataggctg
     3241 gaggagtcct ttttggtgga ggaggataaa aagcacgagc gccacccaat ctttggcaat
     3301 atcgtggacg aggtggcgta ccatgaaaag tacccaacca tatatcatct gaggaagaag
     3361 cttgtagaca gtactgataa ggctgacttg cggttgatct atctcgcgct ggcgcatatg
     3421 atcaaatttc ggggacactt cctcatcgag ggggacctga acccagacaa cagcgatgtc
     3481 gacaaactct ttatccaact ggttcagact tacaatcagc ttttcgaaga gaacccgatc
     3541 aacgcatccg gagttgacgc caaagcaatc ctgagcgcta ggctgtccaa atcccggcgg
     3601 ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga acggcctgtt tggtaatctt
     3661 atcgccctgt cactcgggct gacccccaac tttaaatcta acttcgacct ggccgaagat
     3721 gccaagcttc aactgagcaa agacacctac gatgatgatc tcgacaatct gctggcccag
     3781 atcggcgacc agtacgcaga cctttttttg gcggcaaaga acctgtcaga cgccattctg
     3841 ctgagtgata ttctgcgagt gaacacggag atcaccaaag ctccgctgag cgctagtatg
     3901 atcaagcgct atgatgagca ccaccaagac ttgactttgc tgaaggccct tgtcagacag
     3961 caactgcctg agaagtacaa ggaaattttc ttcgatcagt ctaaaaatgg ctacgccgga
     4021 tacattgacg gcggagcaag ccaggaggaa ttttacaaat ttattaagcc catcttggaa
     4081 aaaatggacg gcaccgagga gctgctggta aagcttaaca gagaagatct gttgcgcaaa
     4141 cagcgcactt tcgacaatgg aagcatcccc caccagattc acctgggcga actgcacgct
     4201 atcctcaggc ggcaagagga tttctacccc tttttgaaag ataacaggga aaagattgag
     4261 aaaatcctca catttcggat accctactat gtaggccccc tcgcccgggg aaattccaga
     4321 ttcgcgtgga tgactcgcaa atcagaagag accatcactc cctggaactt cgaggaagtc
     4381 gtggataagg gggcctctgc ccagtccttc atcgaaagga tgactaactt tgataaaaat
     4441 ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt acgagtactt cacagtttat
     4501 aacgagctca ccaaggtcaa atacgtcaca gaagggatga gaaagccagc attcctgtct
     4561 ggagagcaga agaaagctat cgtggacctc ctcttcaaga cgaaccggaa agttaccgtg
     4621 aaacagctca aagaagacta tttcaaaaag attgaatgtt tcgactctgt tgaaatcagc
     4681 ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc acgatctcct gaaaatcatt
     4741 aaagacaagg acttcctgga caatgaggag aacgaggaca ttcttgagga cattgtcctc
     4801 acccttacgt tgtttgaaga tagggagatg attgaagaac gcttgaaaac ttacgctcat
     4861 ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc gatatacagg atgggggcgg
     4921 ctgtcaagaa aactgatcaa tgggatccga gacaagcaga gtggaaagac aatcctggat
     4981 tttcttaagt ccgatggatt tgccaaccgg aacttcatgc agttgatcca tgatgactct
     5041 ctcaccttta aggaggacat ccagaaagca caagtttctg gccaggggga cagtcttcac
     5101 gagcacatcg ctaatcttgc aggtagccca gctatcaaaa agggaatact gcagaccgtt
     5161 aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata agcccgagaa tatcgttatc
     5221 gagatggccc gagagaacca aactacccag aagggacaga agaacagtag ggaaaggatg
     5281 aagaggattg aagagggtat aaaagaactg gggtcccaaa tccttaagga acacccagtt
     5341 gaaaacaccc agcttcagaa tgagaagctc tacctgtact acctgcagaa cggcagggac
     5401 atgtacgtgg atcaggaact ggacatcaat cggctctccg actacgacgt ggatcatatc
     5461 gtgccccagt cttttctcaa agatgattct attgataata aagtgttgac aagatccgat
     5521 aaaaatagag ggaagagtga taacgtcccc tcagaagaag ttgtcaagaa aatgaaaaat
     5581 tattggcggc agctgctgaa cgccaaactg atcacacaac ggaagttcga taatctgact
     5641 aaggctgaac gaggtggcct gtctgagttg gataaagccg gcttcatcaa aaggcagctt
     5701 gttgagacac gccagatcac caagcacgtg gcccaaattc tcgattcacg catgaacacc
     5761 aagtacgatg aaaatgacaa actgattcga gaggtgaaag ttattactct gaagtctaag
     5821 ctggtctcag atttcagaaa ggactttcag ttttataagg tgagagagat caacaattac
     5881 caccatgcgc atgatgccta cctgaatgca gtggtaggca ctgcacttat caaaaaatat
     5941 cccaagcttg aatctgaatt tgtttacgga gactataaag tgtacgatgt taggaaaatg
     6001 atcgcaaagt ctgagcagga aataggcaag gccaccgcta agtacttctt ttacagcaat
     6061 attatgaatt ttttcaagac cgagattaca ctggccaatg gagagattcg gaagcgacca
     6121 cttatcgaaa caaacggaga aacaggagaa atcgtgtggg acaagggtag ggatttcgcg
     6181 acagtccgga aggtcctgtc catgccgcag gtgaacatcg ttaaaaagac cgaagtacag
     6241 accggaggct tctccaagga aagtatcctc ccgaaaagga acagcgacaa gctgatcgca
     6301 cgcaaaaaag attgggaccc caagaaatac ggcggattcg attctcctac agtcgcttac
     6361 agtgtactgg ttgtggccaa agtggagaaa gggaagtcta aaaaactcaa aagcgtcaag
     6421 gaactgctgg gcatcacaat catggagcga tcaagcttcg aaaaaaaccc catcgacttt
     6481 ctcgaggcga aaggatataa agaggtcaaa aaagacctca tcattaagct tcccaagtac
     6541 tctctctttg agcttgaaaa cggccggaaa cgaatgctcg ctagtgcggg cgagctgcag
     6601 aaaggtaacg agctggcact gccctctaaa tacgttaatt tcttgtatct ggccagccac
     6661 tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga agcagctgtt cgtggaacaa
     6721 cacaaacact accttgatga gatcatcgag caaataagcg aattctccaa aagagtgatc
     6781 ctcgccgacg ctaacctcga taaggtgctt tctgcttaca ataagcacag ggataagccc
     6841 atcagggagc aggcagaaaa cattatccac ttgtttactc tgaccaactt gggcgcgcct
     6901 gcagccttca agtacttcga caccaccata gacagaaagc ggtacacctc tacaaaggag
     6961 gtcctggacg ccacactgat tcatcagtca attacggggc tctatgaaac aagaatcgac
     7021 ctctctcagc tcggtggaga cagcagggct gaccccaaga agaagaggaa ggtgtaatct
     7081 agataattct accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta
     7141 gcagccccgc tgggcacttg gcgctacaca agtggcctct ggcctcgcac acattccaca
     7201 tccaccggta ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact
     7261 cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa
     7321 atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag
     7381 cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag
     7441 aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc
     7501 gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg
     7561 ctgttctcct cttcctcatc tccgggcctt tcgacctgca gcccaagctt accatgaccg
     7621 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     7681 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     7741 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     7801 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     7861 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     7921 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     7981 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     8041 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     8101 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     8161 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgaccgcgtc
     8221 tggaacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     8281 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     8341 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     8401 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     8461 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     8521 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     8581 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     8641 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     8701 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     8761 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga
     8821 attaattctg cagtcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag
     8881 ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag
     8941 tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     9001 ttttaaaaga aaagagggga ctggaagggc taattcactc ccaacgaaga caagatatcc
     9061 ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag
     9121 ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc
     9181 cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc
     9241 tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag
     9301 catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag
     9361 cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg
     9421 gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct
     9481 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     9541 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     9601 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     9661 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     9721 gagaggcctt gacattgcta gcgttttacc gtcgacctct agctagagct tggcgtaatc
     9781 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     9841 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     9901 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     9961 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
    10021 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
    10081 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
    10141 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
    10201 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
    10261 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
    10321 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
    10381 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
    10441 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
    10501 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
    10561 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
    10621 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
    10681 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
    10741 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
    10801 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
    10861 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
    10921 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
    10981 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
    11041 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
    11101 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
    11161 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
    11221 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
    11281 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
    11341 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
    11401 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
    11461 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
    11521 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
    11581 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
    11641 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
    11701 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
    11761 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
    11821 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
    11881 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
    11941 cgacggatcg ggagatcaac ttgtttattg cagcttataa tggttacaaa taaagcaata
    12001 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
    12061 aactcatcaa tgtatcttat catgtctgga tcaactggat aactcaagct aaccaaaatc
    12121 atcccaaact tcccacccca taccctatta ccactgccaa ttacctgtgg tttcatttac
    12181 tctaaacctg tgattcctct gaattatttt cattttaaag aaattgtatt tgttaaatat
    12241 gtactacaaa cttagtagt
//
