LOCUS       .                       6818 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     CDS             95..895
                     /label="ApmR"
                     /note="aminoglycoside 3-N-acetyltransferase type IV"
     terminator      925..1019
                     /label="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     rep_origin      1133..1678
                     /label="p15A ori"
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells
                     that contain a second plasmid with the ColE1 origin."
     terminator      complement(1840..1867)
                     /label="T7Te terminator"
                     /note="phage T7 early transcription terminator"
     terminator      complement(1883..1954)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(1981..6084)
                     /label="dCas9"
                     /note="catalytically dead mutant of the Cas9 endonuclease
                     from the Streptococcus pyogenes Type II CRISPR/Cas system"
     promoter        complement(6120..6175)
                     /label="tetR/tetA promoters"
                     /note="overlapping promoters for bacterial tetR and tetA"
     CDS             6191..6814
                     /label="TetR"
                     /note="tetracycline repressor TetR"
ORIGIN
        1 gggttcatgt gcagctccat cagcaaaagg ggatgataag tttatcacca ccgactattt
       61 gcaacagtgc cgttgatcgt gctatgatcg actgatgtca tcagcggtgg agtgcaatgt
      121 cgtgcaatac gaatggcgaa aagccgagct catcggtcag cttctcaacc ttggggttac
      181 ccccggcggt gtgctgctgg tccacagctc cttccgtagc gtccggcccc tcgaagatgg
      241 gccacttgga ctgatcgagg ccctgcgtgc tgcgctgggt ccgggaggga cgctcgtcat
      301 gccctcgtgg tcaggtctgg acgacgagcc gttcgatcct gccacgtcgc ccgttacacc
      361 ggaccttgga gttgtctctg acacattctg gcgcctgcca aatgtaaagc gcagcgccca
      421 tccatttgcc tttgcggcag cggggccaca ggcagagcag atcatctctg atccattgcc
      481 cctgccacct cactcgcctg caagcccggt cgcccgtgtc catgaactcg atgggcaggt
      541 acttctcctc ggcgtgggac acgatgccaa cacgacgctg catcttgccg agttgatggc
      601 aaaggttccc tatggggtgc cgagacactg caccattctt caggatggca agttggtacg
      661 cgtcgattat ctcgagaatg accactgctg tgagcgcttt gccttggcgg acaggtggct
      721 caaggagaag agccttcaga aggaaggtcc agtcggtcat gcctttgctc ggttgatccg
      781 ctcccgcgac attgtggcga cagccctggg tcaactgggc cgagatccgt tgatcttcct
      841 gcatccgcca gaggcgggat gcgaagaatg cgatgccgct cgccagtcga ttggctgagc
      901 tcattttgat atcgagctcg cttggactcc tgttgataga tccagtaatg acctcagaac
      961 tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc
     1021 caagcactag taacaactta tatcgtatgg ggctgacttc aggtgctaca tttgaagaga
     1081 taaattgcac tgaaatctag aaatatttta tctgattaat aagatgatct tcttgagatc
     1141 gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt
     1201 tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt ggaggagcgc
     1261 agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc aagactaact
     1321 cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg
     1381 gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg gggttcgtgc
     1441 atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg tggaatgaga
     1501 caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa caggagagcg
     1561 cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1621 ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa
     1681 cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc ttccaggaaa
     1741 tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga gcgtagcgag
     1801 tcagtgagcg aggaagcgga atatatccct aggtataaac gcagaaaggc ccacccgaag
     1861 gtgagccagt gtgactctag tagagagcgt tcaccgacaa acaacagata aaacgaaagg
     1921 cccagtcttt cgactgagcc tttcgtttta tttgatgcct ggagatcctt actcgagtta
     1981 gtcacctcct agctgactca aatcaatgcg tgtttcataa agaccagtga tggattgatg
     2041 gataagagtg gcatctaaaa cttcttttgt agacgtatat cgtttacgat caattgttgt
     2101 atcaaaatat ttaaaagcag cgggagctcc aagattcgtc aacgtaaata aatgaataat
     2161 attttctgct tgttcacgta ttggtttgtc tctatgtttg ttatatgcac taagaacttt
     2221 atctaaattg gcatctgcta aaataacacg cttagaaaat tcactgattt gctcaataat
     2281 ctcatctaaa taatgcttat gctgctccac aaacaattgt ttttgttcgt tatcttctgg
     2341 actacccttc aacttttcat aatgactagc taaatataaa aaattcacat atttgcttgg
     2401 cagagccagc tcatttcctt tttgtaattc tccggcacta gccagcatcc gtttacgacc
     2461 gttttctaac tcaaaaagac tatatttagg tagtttaatg attaagtctt ttttaacttc
     2521 cttatatcct ttagcttcta aaaagtcaat cggatttttt tcaaaggaac ttctttccat
     2581 aattgtgatc cctagtaact ctttaacgga ttttaacttc ttcgatttcc ctttttccac
     2641 cttagcaacc actaggactg aataagctac cgttggacta tcaaaaccac catatttttt
     2701 tggatcccag tcttttttac gagcaataag cttgtccgaa tttctttttg gtaaaattga
     2761 ctccttggag aatccgcctg tctgtacttc tgttttcttg acaatattga cttggggcat
     2821 ggacaatact ttgcgcactg tggcaaaatc tcgcccttta tcccagacaa tttctccagt
     2881 ttccccatta gtttcgatta gagggcgttt gcgaatctct ccatttgcaa gtgtaatttc
     2941 tgttttgaag aagttcatga tattagagta aaagaaatat tttgcggttg ctttgcctat
     3001 ttcttgctca gacttagcaa tcattttacg aacatcataa actttataat caccatagac
     3061 aaactccgat tcaagttttg gatatttctt aatcaaagca gttccaacga cggcatttag
     3121 atacgcatca tgggcatgat ggtaattgtt aatctcacgt actttataga attggaaatc
     3181 ttttcggaag tcagaaacta atttagattt taaggtaatc actttaacct ctcgaataag
     3241 tttatcattt tcatcgtatt tagtattcat gcgactatcc aaaatttgtg ccacatgctt
     3301 agtgatttgg cgagtttcaa ccaattggcg tttgataaaa ccagctttat caagttcact
     3361 caaacctcca cgttcagctt tcgttaaatt atcaaactta cgttgagtga ttaacttggc
     3421 gtttagaagt tgtctccaat agtttttcat ctttttgact acttcttcac ttggaacgtt
     3481 atccgattta ccacgatttt tatcagaacg cgttaagacc ttattgtcta ttgaatcgtc
     3541 tttaaggaaa ctttgtggaa caatggcatc gacatcataa tcacttaaac gattaatatc
     3601 taattcttgg tccacataca tgtctcttcc attttggaga taatagagat agagcttttc
     3661 attttgcaat tgagtatttt caacaggatg ctctttaaga atctgacttc ctaattcttt
     3721 gataccttct tcgattcgtt tcatacgctc tcgcgaattt ttctggccct tttgagttgt
     3781 ctgattttca cgtgccattt caataacgat attttctggc ttatgccgcc ccattacttt
     3841 gaccaattca tcaacaactt ttacagtctg taaaatacct tttttaatag cagggctacc
     3901 agctaaattt gcaatatgtt catgtaaact atcgccttgt ccagacactt gtgctttttg
     3961 aatgtcttct ttaaatgtca aactatcatc atggatcagc tgcataaaat tgcgattggc
     4021 aaaaccatct gatttcaaaa aatctaatat tgttttgcca gattgcttat ccctaatacc
     4081 attaatcaat tttcgagaca aacgtcccca accagtataa cggcgacgtt taagctgttt
     4141 catcacctta tcatcaaaga ggtgagcata tgttttaagt ctttcctcaa tcatctccct
     4201 atcttcaaat aaggtcaatg ttaaaacaat atcctctaag atatcttcat tttcttcatt
     4261 atccaaaaaa tctttatctt taataatttt tagcaaatca tggtaggtac ctaatgaagc
     4321 attaaatcta tcttcaactc ctgaaatttc aacactatca aaacattcta tttttttgaa
     4381 ataatcttct tttaattgct taacggttac ttttcgattt gttttgaaga gtaaatcaac
     4441 aatggctttc ttctgttcac ctgaaagaaa tgctggtttt cgcattcctt cagtaacata
     4501 tttgaccttt gtcaattcgt tataaaccgt aaaatactca taaagcaaac tatgttttgg
     4561 tagtactttt tcatttggaa gatttttatc aaagtttgtc atgcgttcaa taaatgattg
     4621 agctgaagca cctttatcga caacttcttc aaaattccat ggggtaattg tttcttcaga
     4681 cttccgagtc atccatgcaa aacgactatt gccacgcgcc aatggaccaa cataataagg
     4741 aattcgaaaa gtcaagattt tttcaatctt ctcacgattg tcttttaaaa atggataaaa
     4801 gtcttcttgt cttctcaaaa tagcatgcag ctcacccaag tgaatttgat ggggaataga
     4861 gccgttgtca aaggtccgtt gcttgcgcag caaatcttca cgatttagtt tcaccaataa
     4921 ttcctcagta ccatccattt tttctaaaat tggtttgata aatttataaa attcttcttg
     4981 gctagctccc ccatcaatat aacctgcata tccgtttttt gattgatcaa aaaagatttc
     5041 tttatacttt tctggaagtt gttgtcgaac taaagctttt aaaagagtca agtcttgatg
     5101 atgttcatcg tagcgtttaa tcattgaagc tgatagggga gccttagtta tttcagtatt
     5161 tactcttagg atatctgaaa gtaaaatagc atctgataaa ttcttagctg ccaaaaacaa
     5221 atcagcatat tgatctccaa tttgcgccaa taaattatct aaatcatcat cgtaagtatc
     5281 ttttgaaagc tgtaatttag catcttctgc caaatcaaaa tttgatttaa aattaggggt
     5341 caaacccaat gacaaagcaa tgagattccc aaataagcca tttttcttct caccggggag
     5401 ctgagcaatg agattttcta atcgtcttga tttactcaat cgtgcagaaa gaatcgcttt
     5461 agcatctact ccacttgcgt taatagggtt ttcttcaaat aattgattgt aggtttgtac
     5521 caactggata aatagtttgt ccacatcact attatcagga tttaaatctc cctcaatcaa
     5581 aaaatgacca cgaaacttaa tcatatgcgc taaggccaaa tagattaagc gcaaatccgc
     5641 tttatcagta gaatctacca atttttttcg cagatgatag atagttggat atttctcatg
     5701 ataagcaact tcatctacta tatttccaaa aataggatga cgttcatgct tcttgtcttc
     5761 ttccaccaaa aaagactctt caagtcgatg aaagaaacta tcatctactt tcgccatctc
     5821 atttgaaaaa atctcctgta gataacaaat acgattcttc cgacgtgtat accttctacg
     5881 agctgtccgt ttgagacgag tcgcttccgc tgtctctcca ctgtcaaata aaagagcccc
     5941 tataagattt tttttgatac tgtggcggtc tgtatttccc agaaccttga actttttaga
     6001 cggaacctta tattcatcag tgatcaccgc ccatccgacg ctatttgtgc cgatagctaa
     6061 gcctattgag tatttcttat ccatagatcc tttctcctct ttagatcttt tgaattcttt
     6121 tctctatcac tgatagggag tggtaaaata actctatcaa cgatagagtg tcaacaaaaa
     6181 ttaggaatta atgatgtcta gattagataa aagtaaagtg attaacagcg cattagagct
     6241 gcttaatgag gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt
     6301 agagcagcct acattgtatt ggcatgtaaa aaataagcgg gctttgctcg acgccttagc
     6361 cattgagatg ttagataggc accatactca cttttgccct ttagaagggg aaagctggca
     6421 agatttttta cgtaataacg ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg
     6481 agcaaaagta catttaggta cacggcctac agaaaaacag tatgaaactc tcgaaaatca
     6541 attagccttt ttatgccaac aaggtttttc actagagaat gcattatatg cactcagcgc
     6601 tgtggggcat tttactttag gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga
     6661 agaaagggaa acacctacta ctgatagtat gccgccatta ttacgacaag ctatcgaatt
     6721 atttgatcac caaggtgcag agccagcctt cttattcggc cttgaattga tcatatgcgg
     6781 attagaaaaa caacttaaat gtgaaagtgg gtcttaag
//
