LOCUS       sequence-66434-e        8899 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gram-Positive Bacterial Shuttle Vector for Nisin-Controlled 
            Inducible Expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8899)
  AUTHORS   Bryan EM, Bae T, Kleerebezem M, Dunny GM
  TITLE     Improved vectors for nisin-controlled expression in gram-positive 
            bacteria.
  JOURNAL   Plasmid. 2000 Sep;44(2):183-90.
  PUBMED    10964628
REFERENCE   2  (bases 1 to 8899)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid."; 
            date: "2000-09"; volume: "44(2)"; pages: "183-90"
COMMENT     Sequence Label: pMSP3535VA
FEATURES             Location/Qualifiers
     source          1..8899
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             536..1330
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      7433..8021
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7922..7941
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8175..8192
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     8279..8298
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8279..8297
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cgataaaccc agcgaaccat ttgaggtgat aggtaagatt ataccgaggt atgaaaacga
       61 gaattggacc tttacagaat tactctatga agcgccatat ttaaaaagct accaagacga
      121 agaggatgaa gaggatgagg aggcagattg ccttgaatat attgacaata ctgataagat
      181 aatatatctt ttatatagaa gatatcgccg tatgtaagga tttcaggggg caaggcatag
      241 gcagcgcgct tatcaatata tctatagaat gggcaaagca taaaaacttg catggactaa
      301 tgcttgaaac ccaggacaat aaccttatag cttgtaaatt ctatcataat tgtggtttca
      361 aaatcggctc cgtcgatact atgttatacg ccaactttca aaacaacttt gaaaaagctg
      421 ttttctggta tttaaggttt tagaatgcaa ggaacagtga attggagttc gtcttgttat
      481 aattagcttc ttggggtatc tttaaatact gtagaaaaga ggaaggaaat aataaatggc
      541 taaaatgaga atatcaccgg aattgaaaaa actgatcgaa aaataccgct gcgtaaaaga
      601 tacggaagga atgtctcctg ctaaggtata taagctggtg ggagaaaatg aaaacctata
      661 tttaaaaatg acggacagcc ggtataaagg gaccacctat gatgtggaac gggaaaagga
      721 catgatgcta tggctggaag gaaagctgcc tgttccaaag gtcctgcact ttgaacggca
      781 tgatggctgg agcaatctgc tcatgagtga ggccgatggc gtcctttgct cggaagagta
      841 tgaagatgaa caaagccctg aaaagattat cgagctgtat gcggagtgca tcaggctctt
      901 tcactccatc gacatatcgg attgtcccta tacgaatagc ttagacagcc gcttagccga
      961 attggattac ttactgaata acgatctggc cgatgtggat tgcgaaaact gggaagaaga
     1021 cactccattt aaagatccgc gcgagctgta tgatttttta aagacggaaa agcccgaaga
     1081 ggaacttgtc ttttcccacg gcgacctggg agacagcaac atctttgtga aagatggcaa
     1141 agtaagtggc tttattgatc ttgggagaag cggcagggcg gacaagtggt atgacattgc
     1201 cttctgcgtc cggtcgatca gggaggatat cggggaagaa cagtatgtcg agctattttt
     1261 tgacttactg gggatcaagc ctgattggga gaaaataaaa tattatattt tactggatga
     1321 attgttttag tacctagatt tagatgtcta aaaagcttta actacaagct ttttagacat
     1381 ctaatctttt ctgaagtaca tccgcaactg tccatactct gatgttttat atcttttcta
     1441 aaagttcgct agataggggt cccgagcgcc tacgaggaat ttgtatcgat tctgagacct
     1501 ctgagaggct ctcagagcta tctaaagctg agggatatat aaatacctta gaaaattatt
     1561 cgaagagctt agaagcgaaa atagagcgtt tagagcgcga ggggctgaaa ttagaaaaac
     1621 taaaaacaca aatagctgac ctaaaaatca tgtctgagaa agaactagcg gctattaccc
     1681 ctaaaaaagg cgtgttcggt aaagaatatg tggaattgac taaagagcag tttgaagaat
     1741 ttaaagggct gatataccga agcagaaacc ttgttcatca aaaagagcta gagaatgagc
     1801 aattaaggcg gatagtgcct ctgagacgct ctaaacggtt tgaagcgagt tggaacgagc
     1861 taaagaaaaa agtaagggag agagcataga gcgtcttagg aacgaaaata gagcgcttag
     1921 aagtgaaaac tcagttttga gacagcaaaa tgacaaaatg ctaggaaaac tgaaagagtt
     1981 tatgccagat aaagccttaa aaaattttat atcagagtta agagctattc agccaatcgt
     2041 aagggtagtt aaacgagtga ttgaaaaagg gctaggcctt tgagcgattt atgccgtgaa
     2101 agctaattga caataagcaa gggcaaagta cgctaggacg tgacgagccg aaaggcttta
     2161 gcgtttcgag ccgacacgga caaaggacgt ccgcccttgg ttacttgttg tcaattagac
     2221 catggaataa agtaagcgga catggtataa tagctaggtc gcaacgttct ttcgctaagt
     2281 tacgaactta gattggaggt gagcgctgtg aagactttcc tagaacttgt tttgatacct
     2341 tttgtggttg gcgttgctgc agaggtaagt gctgattggt tgattcggta tgttcgaaac
     2401 aaacgcgaca aaaattaacc tgagttcttt ttgaggacaa aaaagaaaga cagtagttcc
     2461 agctactgtt ttttttgcgt tgtgctattc gtttcctaga acttctagcg ttaaaattat
     2521 tataccacgt ttggatttag aaagtcaaaa tttgaggttt taggggtgaa tttttcgtga
     2581 aacgaaaaag agggctgaaa agcccttaaa aacccaattg cgtagcaagg gttttttctt
     2641 atcttgatac atatagaaat aatgagtttt tttattttct tgttttaaag cacctcaaac
     2701 ccttgatatt gctgggtttt taggtaacaa aaaagccctt gcaatttaat aaaataaagt
     2761 ttataattta agtgtccaat ttaaaataat aaacttagga gaattgcagg aactttttta
     2821 tatactcaaa aaaatttttt tgcaagaaaa ttataacatg acaggtactg aaaatcaagt
     2881 ctttaaggac tattctaaga atggcaaaga tagaaagtgg cgagaacgca agttaaaaaa
     2941 tattgagctt gctggtcgtt tagagcgttt aggatatcgt tcgtttgaac gggtctatca
     3001 atgtgccgaa gtgttgaagt ttatcgaaca acaagacggc acgaaaaaac tctatcagtc
     3061 ttatttttgc aaaaataagc tctgcccaat ttgtaactgg agacggtcaa tgaagtatgc
     3121 ttatcaagct gaattagtgg taaatgaagc aatgaaacgc tatccaaaag gtcgctttct
     3181 ctttttgacc ttgacgatta agaacattag tggtgaaaaa ttaaataaat caatttcaga
     3241 aatagggcga gcttttaatc gtcttatgaa atacaagaaa gtcgataaaa atgttattgg
     3301 ctatttgcga gccactgagg taacttattc aactgagcat gagaattatc accctcattt
     3361 gcatgtattg ttatttgtga aatctagcta ttttactgga aataatacaa attacattag
     3421 ccaagaagaa tggacgaaac tatgggctaa ggcgatgaaa ttggattata cacctgtagt
     3481 tgatattcga accgtcaaag ctcataaacg taaaaacttg aagtcagcta ttatcgaaac
     3541 agctaaatat cctgttaagc cttttgatgt agatacagaa gatgtgacat tattctctga
     3601 aatggtcaaa gaacggataa cagaagattt aacgaatggt ttgcaccgaa aaaggcagat
     3661 tggttttgga aagttgttca agaaaatcaa ggcggagtta gctcttgatg atgtcgaaga
     3721 agggaatctt gttcagaccg gagcagaaga atctgcagaa agtactcata tcaatacttg
     3781 agggcaatat tggaaaccaa aaaacggaaa ctaaaacgga aacaaaaaca ttagtttcca
     3841 atatgtcttt aagctcaagt gagataaggg aagaagacca tattggaaat cgaaaaatgg
     3901 aaaccaaatt ggaaactgaa ttagaaacta gtttccgata tgtccgtgaa cagttgaaaa
     3961 taaaggaaaa agaggttgaa caacttcatc aattactaga ccaatcacaa cagttacttt
     4021 tgaatgagca gaagaaaaat cagttactgc tagaaatgaa agctgaactt gctgattggg
     4081 aaaaagaaaa aacggagtta gagaagcaag tatcggttta tcaaacttcg ttatctgaag
     4141 ccaatagaag agctaatagc aattatgatt atgctgaaaa tgtagaacgc tggctttgga
     4201 ttgctgggat aactgctggg atattttttg tgattgttct agtcttgatt tggattcact
     4261 ttttttgagt ataagcaagc tgacctttcg gggtcagctt ttttgtccga caatagcgaa
     4321 gcggtcgggc agtcattttg aaaaaatgac attgggcgac ttattgaagt tgcccaatag
     4381 acaaaaatac ccatgtggtt cgagggtgtt tttgctagct cgtcaccttt tctttgtggc
     4441 gaggtaagcc gtctgcaaga cccttcggag aacgcacctt tactatgtaa aggctagtac
     4501 gttacacttt acatggaagt tacaccggaa ttgtgctaaa ctaccagtta agatgttgct
     4561 gattaagacg agcaatagac tgtgtcggct cgtctgtctt tcggtaatcg aaagacagtt
     4621 tttggggagg gaaatctatg agttacatgg ttgcaagaat gcagaagatg aaagctggca
     4681 atttgggagg agcttttaag cataatgaac gtgtttttga gacgcactca aataaagata
     4741 tagacccgag caggtcgcac ctcaactatg agtttccccg gctttaggta tagtgtgtat
     4801 ctcaatcctt ggtatattga aaagaaagac taaaaattga tagattatat ttcttcagaa
     4861 tgaatggtat aatgaagtaa tgagtactaa acaatcggag gtaaagtggt gtataaaatt
     4921 ttaatagttg atgatgatca ggaaatttta aaattaatga agacagcatt agaaatgaga
     4981 aactatgaag ttgcgacgca tcaaaacatt tcacttccct tggatattac tgattttcag
     5041 ggatttgatt tgattttgtt agatatcatg atgtcaaata ttgaagggac agaaatttgt
     5101 aaaaggattc gcagagaaat atcaactcca attatctttg ttagtgcgaa agatacagaa
     5161 gaggatatta taaacggctt aggtattggt ggggatgact atattactaa gccttttagc
     5221 cttaaacagt tggttgcaaa agtggaagca aatataaagc gagaggaacg caataaacat
     5281 gcagttcatg ttttttcaga gattcgtaga gatttaggac caattacatt ttatttagaa
     5341 gaaaggcgag tctgtgtcaa tggtcaaaca attccactga cttgtcgtga atacgatatt
     5401 cttgaattac tatcacaacg aacttctaaa gtttatacga gagaggatat ttatgatgac
     5461 gtatatgatg aatattctaa tgcacttttt cggtcaatct cggagtatat ttatcagatt
     5521 aggagtaagt ttgcaccata cgatattaat ccgataaaaa cggttcgggg acttgggtat
     5581 cagtggcatg ggtaaaaaat attcaatgcg tcgacggata tggcaagctg tcattgaaat
     5641 tatcataggt acttgtctac ttatcctgtt gttactgggc ttgactttct ttctacgaca
     5701 aattggacaa atcagtggtt cagaaactat tcgtttatct ttagattcag ataatttaac
     5761 tatttctgat atcgaacgtg atatgaaaca ctacccatat gattatatta tttttgacaa
     5821 tgatacaagt aaaattttgg gaggacatta tgtcaagtcg gatgtaccta gttttgtagc
     5881 ttcaaaacag tcttcacata atattacaga aggagaaatt acttatactt attcaagcaa
     5941 taagcatttt tcagttgttt taagacaaaa cagtatgcct gaatttacaa atcatacgct
     6001 tcgttcaatt tcttataatc aatttactta ccttttcttt tttcttggtg aaataatact
     6061 cattattttt tctgtctatc atctcattag agaattttct aagaattttc aagccgttca
     6121 aaagattgca ttgaagatgg gggaaataac tacttttcct gaacaagagg aatcaaaaat
     6181 tattgaattt gatcaggttc tgaataactt atattcgaaa agtaaggagt tagctttcct
     6241 tattgaagcg gagcgtcatg aaaaacatga tttatccttc caggttgctg cactttcaca
     6301 tgatgttaag acacctttaa cagtattaaa aggaaatatt gaactgctag agatgactga
     6361 agtaaatgaa caacaagctg attttattga gtcaatgaaa aatagtttga ctgtttttga
     6421 caagtatttt aacacaatga ttagttatac aaaacttttg aatgatgaaa atgattacaa
     6481 agcgacaatc tccctggagg attttttgat agatttatca gttgagttgg aagagttgtc
     6541 aacaacttat caagtggatt atcagctagt taaaaaaaca gatttaacca ctttttacgg
     6601 aaatacatta gctttaagtc gagcacttat caatatcttt gttaatgcct gtcagtatgc
     6661 taaagagggt gaaaaaatag tcagtttgag tatttatgat gatgaaaaat atctctattt
     6721 tgaaatctgg aataatggtc atcctttttc tgaacaagca aaaaaaaatg ctggaaaact
     6781 atttttcaca gaagatactg gacgtagtgg gaaacactat gggattggac tatcttttgc
     6841 tcaaggtgta gctttaaaac atcaaggaaa cttaattctc agtaatcctc aaaaaggtgg
     6901 ggcagaagtt atcctaaaaa taaaaaagta atttagtaat ctctaaggat tacttttttt
     6961 gtttcggcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     7021 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     7081 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     7141 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     7201 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     7261 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     7321 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     7381 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     7441 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     7501 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     7561 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7621 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7681 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7741 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7801 aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg
     7861 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7921 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7981 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     8041 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     8101 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     8161 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     8221 cgcctctccc cgcgcgttgg ccgattcatt aatgcaggtt aacctggctt atcgaaatta
     8281 atacgactca ctatagggag accggcctcg agtctagaag cgctgtcgac gagctcgcat
     8341 gcgaattcct gcagcccggg actagtggta ccggatccat gcagagtctc ctgttttaca
     8401 accgggtgta catagcgaaa tacttgtaat gcgtggtgat gcacctgaat ctttcttcga
     8461 aacagatacc aaatccaagc taaaatcttt tgtactcatt ttgagtgcct ccttataatt
     8521 tattttgtag ttccttcgaa cgaaatcatt gtatctaaca aacttcagaa tttaatcaga
     8581 gccgtttatt atgctcgcgt tatcgacaat aatattatta ccaatacttt ctcaagatag
     8641 aattaagact gttttagatt tgttaatgtt tctattgtca gtatagttat aagactagat
     8701 ctgatccgta gcggttttca aaatttgcaa ccaggaatga attactatgt gaaacaacta
     8761 accgagaaaa aatggactgg ttaagaaaac agtaccaaga agagctgaaa aaactagaat
     8821 cgtctaaaaa gcccctagaa gacgatttaa gccatttaga agagttgctt gatgaaaaga
     8881 ccaaggaata tattaaaat
//
