LOCUS       Exported                6547 bp DNA     circular SYN 29-JUL-2025
DEFINITION  Exported.
ACCESSION   V014299
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6547)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6547)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6547)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6547)
  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..6547
                     /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
                     /label=PnisA
                     /note="Originated from Lactococcus lactis subsp. Lactis,
                     PnisA is a member of the two-component regulatory system 
                     NisK/NisR involved in the regulation of the biosynthesis of
                     lantibiotic nisin. PnisA functions as a promoter which is 
                     activated by phosphorylated NisR and enhance downstream 
                     gene expression in response to environmental nisin 
                     signals."
     CDS             2444..3160
                     /label=EGFP
                     /note="enhanced GFP"
     CDS             complement(3577..3588)
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     CDS             3722..3931
                     /codon_start=1
                     /label=repC
                     /note="repC"
                     /note="Replication gene C"
                     /translation="MGGKEANFASVLRPPIKCRVPIFVPKTLYPNWLKGLRGFSIANES
                     PTFSPTFFINLYLSSFIFVFMITK"
     CDS             4200..4898
                     /codon_start=1
                     /label=repA
                     /note="repA"
                     /note="Replication gene A"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNSSDVIRNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDRYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     CDS             5358..6005
                     /gene="cat"
                     /label=Chloramphenicol acetyltransferase
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     primer_bind     complement(6153..6169)
                     /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 accatggtga gcaagggcga
     2461 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2521 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2581 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2641 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2701 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     2761 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2821 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     2881 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     2941 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     3001 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     3061 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     3121 cgccgccggg atcactctcg gcatggacga gctgtacaag taactagttc tagagagctc
     3181 aagctttctt tgaaccaaaa ttagaaaacc aaggcttgaa acgttcaatt gaaatggcaa
     3241 ttaaacaaat tacagcacgt gttgctttga ttgatagcca aaaagcagca gttgataaag
     3301 caattactga tattgctgaa aaattgtaat ttataaataa aaatcacctt ttagaggtgg
     3361 tttttttatt tataaattat tcgtttgatt tcgctttcga tagaacaatc aaatcgtttc
     3421 tgagacgttt tagcgtttat ttcgtttagt tatcggcata atcgttaaaa caggcgttat
     3481 cgtagcgtaa aagcccttga gcgtagcgtg gctttgcagc gaagatgttg tctgttagat
     3541 tatgaaagcc gatgactgaa tgaaataata agcgcagcgt ccttctattt cggttggagg
     3601 aggctcaagg gagtttgagg gaatgaaatt ccctcatggg tttgatttta aaaattgctt
     3661 gcaattttgc cgagcggtag cgctggaaaa tttttgaaaa aaatttggaa tttggaaaaa
     3721 aatgggggga aaggaagcga attttgcttc cgtactacga ccccccatta agtgccgagt
     3781 gccaattttt gtgccaaaaa cgctctatcc caactggctc aagggtttga ggggtttttc
     3841 aatcgccaac gaatcgccaa cgttttcgcc aacgtttttt ataaatctat atttaagtag
     3901 ctttattttt gtttttatga ttacaaagtg atacactaat tttataaaat tatttgattg
     3961 gagtttttta aatggtgatt tcagaatcga aaaaaagagt tatgatttct ctgacaaaag
     4021 agcaagataa aaaattaaca gatatggcga aacaaaaaga tttttcaaaa tctgcggttg
     4081 cggcgttagc tatagaagaa tatgcaagaa aggaatcaga acaaaaaaaa taagcgaaag
     4141 ctcgcgtttt tagaaggata cgagttttcg ctacttgttt ttgataaggt aattatatca
     4201 tggctattaa aaatactaaa gctagaaatt ttggattttt attatatcct gactcaattc
     4261 ctaatgattg gaaagaaaaa ttagagagtt tgggcgtatc tatggctgtc agtcctttac
     4321 acgatatgga cgaaaaaaaa gataaagata catggaatag tagtgatgtt atacgaaatg
     4381 gaaagcacta taaaaaacca cactatcacg ttatatatat tgcacgaaat cctgtaacaa
     4441 tagaaagcgt taggaacaag attaagcgaa aattggggaa tagttcagtt gctcatgttg
     4501 agatacttga ttatatcaaa ggttcatatg aatatttgac tcatgaatca aaggacgcta
     4561 ttgctaagaa taaacatata tacgacaaaa aagatatttt gaacattaat gattttgata
     4621 ttgaccgcta tataacactt gatgaaagcc aaaaaagaga attgaagaat ttacttttag
     4681 atatagtgga tgactataat ttggtaaata caaaagattt aatggctttt attcgcctta
     4741 ggggagcgga gtttggaatt ttaaatacga atgatgtaaa agatattgtt tcaacaaact
     4801 ctagcgcctt tagattatgg tttgagggca attatcagtg tggatataga gcaagttatg
     4861 caaaggttct tgatgctgaa acgggggaaa taaaatgaca aacaaagaaa aagagttatt
     4921 tgctgaaaat gaggaattaa aaaaagaaat taaggactta taagagcgta ttgaaagata
     4981 cagagaaatg gaagttgaat taagtacaac aatagattta ttgagaggag ggattattga
     5041 ataaataaaa gcccccctga cgaaagtcga cggcaatagt tacccttatt atcaagataa
     5101 gaaagaaaag gatttttcgc tacgctcaaa tcctttaaaa aaacacaaaa gaccacattt
     5161 tttaatgtgg tcttttattc ttcaactaaa gcacccatta gttcaacaaa cgaaaattgg
     5221 ataaagtggg atatttttaa aatatatatt tatgttacag taatattgac ttttaaaaaa
     5281 ggattgattc taatgaagaa agcagacaag taagcctcct aaattcactt tagataaaaa
     5341 tttaggaggc atatcaaatg aactttaata aaattgattt agacaattgg aagagaaaag
     5401 agatatttaa tcattatttg aaccaacaaa cgacttttag tataaccaca gaaattgata
     5461 ttagtgtttt ataccgaaac ataaaacaag aaggatataa attttaccct gcatttattt
     5521 tcttagtgac aagggtgata aactcaaata cagcttttag aactggttac aatagcgacg
     5581 gagagttagg ttattgggat aagttagagc cactttatac aatttttgat ggtgtatcta
     5641 aaacattctc tggtatttgg actcctgtaa agaatgactt caaagagttt tatgatttat
     5701 acctttctga tgtagagaaa tataatggtt cggggaaatt gtttcccaaa acacctatac
     5761 ctgaaaatgc tttttctctt tctattattc cttggacttc atttactggg tttaacttaa
     5821 atatcaataa taatagtaat taccttctac ccattattac agcaggaaaa ttcattaata
     5881 aaggtaattc aatatattta ccgctatctt tacaggtaca tcattctgtt tgtgatggtt
     5941 atcatgctgg attgtttatg aactctattc aggaattgtc agataggcct aatgactggc
     6001 ttttataata tgagataatg ccgactgtac tttttacagt cggttttcta atgtcactaa
     6061 cctgccccgt tagttgaaga aggtttttat attacagctc cagatctgga tccgatatct
     6121 agatgcattc gcgaggtacc gagctcgaat tcactggccg tcgttttaca acgtcgtgac
     6181 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     6241 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     6301 ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     6361 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     6421 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     6481 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     6541 cgcgcga
//
