LOCUS       Exported                5853 bp DNA     circular SYN 29-JUL-2025
DEFINITION  Exported.
ACCESSION   V013967
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5853)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5853)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5853)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5853)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5853
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..200
                     /label=AmpR promoter
     CDS             201..1058
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1232..1820
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2144..2174
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2245..2428
                     /note="Pnis"
                     /note="Nisin A promoter region"
     terminator      2633..2685
                     /note="terminator"
     CDS             complement(2883..2894)
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     CDS             3028..3237
                     /codon_start=1
                     /label=repC
                     /note="repC"
                     /note="Replication gene C"
                     /translation="MGGKEANFASVLRPPIKCRVPIFVPKTLYPNWLKGLRGFSIANES
                     PTFSPTFFINLYLSSFIFVFMITK"
     CDS             3506..4204
                     /codon_start=1
                     /label=repA
                     /note="repA"
                     /note="Replication gene A"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNSSDVIRNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDRYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     CDS             4664..5311
                     /gene="cat"
                     /label=Chloramphenicol acetyltransferase
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     primer_bind     complement(5459..5475)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt agatctagtc ttataactat actgacaata gaaacattaa
     2281 caaatctaaa acagtcttaa ttctatcttg agaaagtatt ggtaataata ttattgtcga
     2341 taacgcgagc ataataaacg gctctgatta aattctgaag tttgttagat acaatgattt
     2401 cgttcgaagg aactacaaaa taaattataa ggaggcactc accatgggta ctgcaggcat
     2461 gcggtaccac tagttctaga gagctcaagc tttctttgaa ccaaaattag aaaaccaagg
     2521 cttgaaacgt tcaattgaaa tggcaattaa acaaattaca gcacgtgttg ctttgattga
     2581 tagccaaaaa gcagcagttg ataaagcaat tactgatatt gctgaaaaat tgtaatttat
     2641 aaataaaaat caccttttag aggtggtttt tttatttata aattattcgt ttgatttcgc
     2701 tttcgataga acaatcaaat cgtttctgag acgttttagc gtttatttcg tttagttatc
     2761 ggcataatcg ttaaaacagg cgttatcgta gcgtaaaagc ccttgagcgt agcgtggctt
     2821 tgcagcgaag atgttgtctg ttagattatg aaagccgatg actgaatgaa ataataagcg
     2881 cagcgtcctt ctatttcggt tggaggaggc tcaagggagt ttgagggaat gaaattccct
     2941 catgggtttg attttaaaaa ttgcttgcaa ttttgccgag cggtagcgct ggaaaatttt
     3001 tgaaaaaaat ttggaatttg gaaaaaaatg gggggaaagg aagcgaattt tgcttccgta
     3061 ctacgacccc ccattaagtg ccgagtgcca atttttgtgc caaaaacgct ctatcccaac
     3121 tggctcaagg gtttgagggg tttttcaatc gccaacgaat cgccaacgtt ttcgccaacg
     3181 ttttttataa atctatattt aagtagcttt atttttgttt ttatgattac aaagtgatac
     3241 actaatttta taaaattatt tgattggagt tttttaaatg gtgatttcag aatcgaaaaa
     3301 aagagttatg atttctctga caaaagagca agataaaaaa ttaacagata tggcgaaaca
     3361 aaaagatttt tcaaaatctg cggttgcggc gttagctata gaagaatatg caagaaagga
     3421 atcagaacaa aaaaaataag cgaaagctcg cgtttttaga aggatacgag ttttcgctac
     3481 ttgtttttga taaggtaatt atatcatggc tattaaaaat actaaagcta gaaattttgg
     3541 atttttatta tatcctgact caattcctaa tgattggaaa gaaaaattag agagtttggg
     3601 cgtatctatg gctgtcagtc ctttacacga tatggacgaa aaaaaagata aagatacatg
     3661 gaatagtagt gatgttatac gaaatggaaa gcactataaa aaaccacact atcacgttat
     3721 atatattgca cgaaatcctg taacaataga aagcgttagg aacaagatta agcgaaaatt
     3781 ggggaatagt tcagttgctc atgttgagat acttgattat atcaaaggtt catatgaata
     3841 tttgactcat gaatcaaagg acgctattgc taagaataaa catatatacg acaaaaaaga
     3901 tattttgaac attaatgatt ttgatattga ccgctatata acacttgatg aaagccaaaa
     3961 aagagaattg aagaatttac ttttagatat agtggatgac tataatttgg taaatacaaa
     4021 agatttaatg gcttttattc gccttagggg agcggagttt ggaattttaa atacgaatga
     4081 tgtaaaagat attgtttcaa caaactctag cgcctttaga ttatggtttg agggcaatta
     4141 tcagtgtgga tatagagcaa gttatgcaaa ggttcttgat gctgaaacgg gggaaataaa
     4201 atgacaaaca aagaaaaaga gttatttgct gaaaatgagg aattaaaaaa agaaattaag
     4261 gacttataag agcgtattga aagatacaga gaaatggaag ttgaattaag tacaacaata
     4321 gatttattga gaggagggat tattgaataa ataaaagccc ccctgacgaa agtcgacggc
     4381 aatagttacc cttattatca agataagaaa gaaaaggatt tttcgctacg ctcaaatcct
     4441 ttaaaaaaac acaaaagacc acatttttta atgtggtctt ttattcttca actaaagcac
     4501 ccattagttc aacaaacgaa aattggataa agtgggatat ttttaaaata tatatttatg
     4561 ttacagtaat attgactttt aaaaaaggat tgattctaat gaagaaagca gacaagtaag
     4621 cctcctaaat tcactttaga taaaaattta ggaggcatat caaatgaact ttaataaaat
     4681 tgatttagac aattggaaga gaaaagagat atttaatcat tatttgaacc aacaaacgac
     4741 ttttagtata accacagaaa ttgatattag tgttttatac cgaaacataa aacaagaagg
     4801 atataaattt taccctgcat ttattttctt agtgacaagg gtgataaact caaatacagc
     4861 ttttagaact ggttacaata gcgacggaga gttaggttat tgggataagt tagagccact
     4921 ttatacaatt tttgatggtg tatctaaaac attctctggt atttggactc ctgtaaagaa
     4981 tgacttcaaa gagttttatg atttatacct ttctgatgta gagaaatata atggttcggg
     5041 gaaattgttt cccaaaacac ctatacctga aaatgctttt tctctttcta ttattccttg
     5101 gacttcattt actgggttta acttaaatat caataataat agtaattacc ttctacccat
     5161 tattacagca ggaaaattca ttaataaagg taattcaata tatttaccgc tatctttaca
     5221 ggtacatcat tctgtttgtg atggttatca tgctggattg tttatgaact ctattcagga
     5281 attgtcagat aggcctaatg actggctttt ataatatgag ataatgccga ctgtactttt
     5341 tacagtcggt tttctaatgt cactaacctg ccccgttagt tgaagaaggt ttttatatta
     5401 cagctccaga tctggatccg atatctagat gcattcgcga ggtaccgagc tcgaattcac
     5461 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     5521 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     5581 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta
     5641 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     5701 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     5761 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     5821 agaggttttc accgtcatca ccgaaacgcg cga
//
