LOCUS       .                      10552 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        1..204
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             340..360
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             358..4461
                     /label="Cas9(H840A)"
                     /note="nickase mutant of the Cas9 endonuclease from the
                     Streptococcus pyogenes Type II CRISPR/Cas system"
     CDS             4519..4539
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             7318..7338
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             7348..7374
                     /label="c-myc NLS"
                     /note="nuclear localization signal of human c-Myc
                     proto-oncogene (Dang and Lee, 1988)"
     polyA_signal    7482..7706
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(7825..7855)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7870..7891)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(8179..8767)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8941..9798)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(9799..9903)
                     /label="AmpR promoter"
     enhancer        10173..10552
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatctcgag ctcggtacct
      241 aatacgacac actataagga aataagagag aaaagaagag taagaagaaa tataagagcc
      301 accatgaaac ggacagccga cggaagcgag ttcgagtcac caaagaagaa gcggaaagtc
      361 gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc
      421 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
      481 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
      541 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
      601 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
      661 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
      721 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
      781 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
      841 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
      901 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
      961 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagaaag
     1021 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg
     1081 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
     1141 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
     1201 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
     1261 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
     1321 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
     1381 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
     1441 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
     1501 aagatggacg gcaccgagga actgctcgtg aagctgaaga gagaggacct gctgcggaag
     1561 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
     1621 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
     1681 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
     1741 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
     1801 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
     1861 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
     1921 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
     1981 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
     2041 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
     2101 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
     2161 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
     2221 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
     2281 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
     2341 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
     2401 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc
     2461 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
     2521 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
     2581 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
     2641 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
     2701 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
     2761 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
     2821 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggacgctatc
     2881 gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac
     2941 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
     3001 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc
     3061 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
     3121 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
     3181 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag
     3241 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
     3301 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
     3361 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
     3421 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
     3481 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct
     3541 ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc
     3601 accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
     3661 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc
     3721 agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat
     3781 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
     3841 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
     3901 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
     3961 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag
     4021 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
     4081 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
     4141 cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
     4201 ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc
     4261 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
     4321 gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag
     4381 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
     4441 ctgtctcagc tgggaggtga ctccggcgga agctctggtg gcagcaagcg gaccgccgac
     4501 ggctctgaat tcgagagccc taagaagaaa agaaaggtga gcggaggctc tagcggcgga
     4561 agcaccctga acattgaaga cgagtataga ctgcatgaaa caagcaagga acccgatgtg
     4621 tccctgggct ccacctggct gtccgacttt ccccaggcct gggccgagac aggaggaatg
     4681 ggcctggccg tgcggcaggc acccctgatc atccctctga aggccacctc tacacccgtg
     4741 agcatcaagc agtaccctat gtctcaggag gccagactgg gcatcaagcc tcacatccag
     4801 aggctgctgg accagggcat cctggtgcca tgccagagcc cctggaacac accactgctg
     4861 cccgtgaaga agccaggcac caatgactat agacccgtgc aggatctgag agaggtgaac
     4921 aagagggtgg aggatatcca ccccaccgtg cccaaccctt acaatctgct gtccggcctg
     4981 cccccttctc accagtggta tacagtgctg gacctgaagg atgccttctt ttgtctgaga
     5041 ctgcacccta ccagccagcc actgttcgcc tttgagtgga gggaccctga gatgggcatc
     5101 tctggccagc tgacctggac acgcctgcct cagggcttca agaatagccc aacactgttt
     5161 aacgaggccc tgcaccgcga cctggcagat ttccggatcc agcacccaga tctgatcctg
     5221 ctgcagtacg tggacgatct gctgctggcc gccaccagcg agctggattg ccagcaggga
     5281 acacgcgccc tgctgcagac cctgggaaac ctgggatata gggcatccgc caagaaggcc
     5341 cagatctgtc agaagcaggt gaagtacctg ggctatctgc tgaaggaggg ccagagatgg
     5401 ctgacagagg ccaggaagga gacagtgatg ggccagccaa cacccaagac cccaagacag
     5461 ctgagggagt tcctgggcaa agcaggattt tgcaggctgt tcatcccagg attcgcagag
     5521 atggcagcac ctctgtaccc actgaccaag ccgggcaccc tgtttaattg gggccctgac
     5581 cagcagaagg cctatcagga gatcaagcag gccctgctga cagcaccagc cctgggcctg
     5641 ccagacctga ccaagccttt cgagctgttt gtggatgaga agcagggcta cgccaagggc
     5701 gtgctgaccc agaagctggg accatggaga cggcccgtgg cctatctgtc caagaagctg
     5761 gacccagtgg cagcaggatg gccaccatgc ctgaggatgg tggcagcaat cgccgtgctg
     5821 acaaaggatg ccggcaagct gaccatggga cagccactgg tcatcctggc accacacgca
     5881 gtggaggccc tggtgaagca gcctccagat cgctggctgt ctaacgcccg gatgacacac
     5941 taccaggccc tgctgctgga caccgatcgc gtgcagtttg gccctgtggt ggccctgaat
     6001 ccagccaccc tgctgcctct gccagaggag ggcctgcagc acaactgtct ggacatcctg
     6061 gcagaggcac acggaacaag gccagacctg accgatcagc ccctgcctga cgccgatcac
     6121 acatggtata ccgatggaag ctccctgctg caggagggcc agaggaaggc aggagcagca
     6181 gtgaccacag agacagaagt gatctgggcc aaggccctgc cagcaggcac atccgcccag
     6241 cgggccgagc tgatcgccct gacccaggcc ctgaagatgg ccgagggcaa gaagctgaac
     6301 gtgtacacag actccagata tgccttcgcc accgcacaca tccacggaga gatctacagg
     6361 cgccggggct ggctgacctc tgagggcaag gagatcaaga acaaggatga gatcctggcc
     6421 ctgctgaagg ccctgtttct gcccaagcgg ctgagcatca tccactgtcc tggacaccag
     6481 aagggacact ccgccgaggc aaggggcaat cggatggccg accaggccgc cagaaaggct
     6541 gctattactg aaactcccga cacttccact ctgctgattg aaaactcctc cccttccgga
     6601 ggatctagcg gaggctcctc tggctctgag acacctggca caagcgagag cgcaacacct
     6661 gaaagcagcg ggggcagcag cggggggtca atggctgaaa atggtgataa tgaaaagatg
     6721 gctgccctgg aggccaaaat ctgtcatcaa attgagtatt attttggcga cttcaatttg
     6781 ccacgggaca agtttctaaa ggaacagata aaactggatg aaggctgggt acctttggag
     6841 ataatgataa aattcaacag gttgaaccgt ctaacaacag actttaatgt aattgtggaa
     6901 gcattgagca aatccaaggc agaactcatg gaaatcagtg aagataaaac taaaatcaga
     6961 aggtctccaa gcaaacccct acctgaagtg actgatgagt ataaaaatga tgtaaaaaac
     7021 agatctgttt atattaaagg cttcccaact gatgcaactc ttgatgacat aaaagaatgg
     7081 ttagaagata aaggtcaagt actaaatatt cagatgagaa gaacattgca taaagcattt
     7141 aagggatcaa tttttgttgt gtttgatagc attgaatctg ctaagaaatt tgtagagacc
     7201 cctggccaga agtacaaaga aacagacctg ctaatacttt tcaaggacga ttactttgcc
     7261 aaaaaaaatg aatctggcgg ctcaaaaaga accgccgacg gcagcgaatt cgagtctccc
     7321 aagaagaaga ggaaagtcgg ctctggccct gccgctaaga gagtgaagct ggactaatta
     7381 attaagctgc cttctgcggg gcttgccttc tggccatgcc cttcttctct cccttgcacc
     7441 tgtacctctt ggtctttgaa taaagcctga gtaggaagcg actgtgcctt ctagttgcca
     7501 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     7561 tgtcctttcc taataaaatg agaaaattgc atcgcattgt ctgagtaggt gtcattctat
     7621 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     7681 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctcgat
     7741 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     7801 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagccta
     7861 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7921 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7981 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     8041 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     8101 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     8161 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     8221 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     8281 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     8341 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     8401 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     8461 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     8521 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     8581 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     8641 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8701 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     8761 atctcaagaa gatcctttga tcttttctac ggggtctgac actcagtgga acgaaaactc
     8821 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     8881 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     8941 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     9001 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     9061 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     9121 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     9181 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     9241 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     9301 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     9361 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     9421 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     9481 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     9541 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     9601 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     9661 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     9721 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     9781 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     9841 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     9901 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc gatctcccga
     9961 tcccctaggg tctactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
    10021 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
    10081 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
    10141 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
    10201 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
    10261 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
    10321 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
    10381 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
    10441 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
    10501 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tg
//
