LOCUS       V004456                 8353 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004456
VERSION     V004456
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8353)
  AUTHORS   Bryan EM, Bae T, Kleerebezem M, Dunny GM.
  TITLE     Improved vectors for nisin-controlled expression in gram-positive
            bacteria
  JOURNAL   Plasmid 44 (2), 183-190 (2000)
   PUBMED   10964628
REFERENCE   2  (bases 1 to 8353)
  AUTHORS   Staddon JH, Bryan EM, Manias DA, Dunny GM.
  TITLE     Conserved target for group II intron insertion in relaxase genes of
            conjugative elements of gram-positive bacteria
  JOURNAL   J. Bacteriol. 186 (8), 2393-2401 (2004)
   PUBMED   15060042
REFERENCE   3  (bases 1 to 8353)
  AUTHORS   Staddon JH, Bryan EM, Manias DA, Bae T, Kleerebezem M, Dunny GM.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-MAY-2003) Microbiology, University of Minnesota, MMC
            196, 420 Delaware Street S.E., Minneapolis, MN 55455, USA
REFERENCE   4  (bases 1 to 8353)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 8353)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid";
            date: "2000"; volume: "44"; issue: "2"; pages: "183-190"
            SGRef: number: 2; type: "Journal Article"; journalName: "J.
            Bacteriol."; date: "2004"; volume: "186"; issue: "8"; pages:
            "2393-2401"
            SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
            (21-MAY-2003) Microbiology, University of Minnesota, MMC 196, 420
            Delaware Street S.E., Minneapolis, MN 55455, USA"
            SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8353
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             332..1066
                     /gene="ermBP"
                     /label="rRNA adenine N-6-methyltransferase"
                     /note="rRNA adenine N-6-methyltransferase from Enterococcus
                     faecalis. Accession#: P0A4D5"
     CDS             2046..2345
                     /codon_start=1
                     /product="RepD"
                     /label="RepD"
                     /protein_id="AAP60030.1"
                     /translation="MNHDECKTYIKNSLLEIRKLANIYTLETFKKELEKRNIYLETKSD
                     KYFSSEGEDYIYKLIENNKIIYSISGKKLTYKGKKSFSKHAILKQLNEKANQVN"
     CDS             2388..3878
                     /codon_start=1
                     /product="RepE"
                     /label="RepE"
                     /note="plasmid replication; pAMbeta1 plasmid replicon"
                     /protein_id="AAP60031.1"
                     /translation="MNIPFVVETVLHDGLLKYKFKNSKIRSITTKPGKSKGAIFAYRSK
                     SSMIGGRGVVLTSEEAIQENQDTFTHWTPNVYRYGTYADENRSYTKGHSENNLRQINTF
                     FIDFDIHTAKETISASDILTTAIDLGFMPTMIIKSDKGYQAYFVLETPVYVTSKSEFKS
                     VKAAKIISQNIREYFGKSLPVDLTCNHFGIARIPRTDNVEFFDPNYRYSFKEWQDWSFK
                     QTDNKGFTRSSLTVLSGTEGKKQVDEPWFNLLLHETKFSGEKGLIGRNNVMFTLSLAYF
                     SSGYSIETCEYNMFEFNNRLDQPLEEKEVIKIVRSAYSENYQGANREYITILCKAWVSS
                     DLTSKDLFVRQGWFKFKKKRSERQRVHLSEWKEDLMAYISEKSDVYKPYLVTTKKEIRE
                     VLGIPERTLDKLLKVLKANQEIFFKIKPGRNGGIQLASVKSLLLSIIKVKKEEKESYIK
                     ALTNSFDLEHTFIQETLNKLAERPKTDTQLDLFSYDTG"
     CDS             4228..4365
                     /codon_start=1
                     /product="RepG"
                     /label="RepG"
                     /protein_id="AAP60032.1"
                     /translation="MEKYLKNPITWIGLVLVVTWFLTKSSEFLIFGVCVLLLVFASQSD
                     "
     CDS             4614..5252
                     /codon_start=1
                     /product="NisR"
                     /label="NisR"
                     /note="response regulator"
                     /protein_id="AAP60033.1"
                     /translation="MKTALEMRNYEVATHQNISLPLDITDFQGFDLILLDIMMSNIEGT
                     EICKRIRREISTPIIFVSAKDTEEDIINGLGIGGDDYITKPFSLKQLVAKVEANIKREE
                     RNKHAVHVFSEIRRDLGPITFYLEERRVCVNGQTIPLTCREYDILELLSQRTSKVYTRE
                     DIYDDVYDEYSNALFRSISEYIYQIRSKFAPYDINPIKTVRGLGYQWHG"
     CDS             5245..6585
                     /gene="nisK"
                     /label="Nisin biosynthesis sensor protein NisK"
                     /note="Nisin biosynthesis sensor protein NisK from
                     Lactococcus lactis subsp. lactis. Accession#: P42707"
     rep_origin      7090..7678
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     promoter        7936..7954
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      8045..8353
                     /note="PnisA; nisin inducible promoter"
                     /regulatory_class="promoter"
ORIGIN
        1 agatctgatc cgtagcggtt ttcaaaattt gcaaccagga atgaattact atccctttta
       61 tcaagaagcg caaaagaaaa acgaaatgat acaccaatca gtgcaaaaaa agatataatg
      121 ggagataaga cggttcgtgt tcgtgctgac ttgcaccata tcataaaaat cgaaacagca
      181 aagaatggcg gaaacgtaaa agaagttatg gaaataagac ttagaagcaa acttaagagt
      241 gtgttgatag tgcagtatct taaaattttg tataatagga attgaagtta aattagatgc
      301 taaaaatttg taattaagaa ggagtgatta catgaacaaa aatataaaat attctcaaaa
      361 ctttttaacg agtgaaaaag tactcaacca aataataaaa caattgaatt taaaagaaac
      421 cgataccgtt tacgaaattg gaacaggtaa agggcattta acgacgaaac tggctaaaat
      481 aagtaaacag gtaacgtcta ttgaattaga cagtcatcta ttcaacttat cgtcagaaaa
      541 attaaaactg aatactcgtg tcactttaat tcaccaagat attctacagt ttcaattccc
      601 taacaaacag aggtataaaa ttgttgggag tattccttac catttaagca cacaaattat
      661 taaaaaagtg gtttttgaaa gccatgcgtc tgacatctat ctgattgttg aagaaggatt
      721 ctacaagcgt accttggata ttcaccgaac actagggttg ctcttgcaca ctcaagtctc
      781 gattcagcaa ttgcttaagc tgccagcgga atgctttcat cctaaaccaa aagtaaacag
      841 tgtcttaata aaacttaccc gccataccac agatgttcca gataaatatt ggaagctata
      901 tacgtacttt gtttcaaaat gggtcaatcg agaatatcgt caactgttta ctaaaaatca
      961 gtttcatcaa gcaatgaaac acgccaaagt aaacaattta agtaccgtta cttatgagca
     1021 agtattgtct atttttaata gttatctatt atttaacggg aggaaataat tctatgagtc
     1081 gcttttgtaa atttggaaag ttacacgtta ctaaagggaa tgtagataaa ttattaggta
     1141 tactactgac agcttccaag gagctaaaga ggtccctagc gcttagaatc gctttaggaa
     1201 acacgatcca gtccaataat cgtcgataaa aacttttgaa aaaggttggt gaaattacct
     1261 acttttggaa taatcacaaa tcacaagtga ttaatcacaa atcacaagtg attaatcact
     1321 tgtttattaa gatattaaaa gctataattt aaataaagcg tgaattttat tacacaaaaa
     1381 gaggggggag aaacttggaa ctagcattta gagaaagctt aaaaaagatg agaggtacca
     1441 aatcaaaaga aaaattctcc caagaattag aaatgagtag atcaaattat tcacgaatag
     1501 aatcaggaaa atcagatcca accataaaaa cactagaaca aattgcaaag ttaactaact
     1561 caacgctagt agtggattta atcccaaatg agccaacaga accagaacca gaaacagaat
     1621 cagaacaagt aacattggat ttagaaatgg aagaagaaaa aagcaatgac ttcgtgtgaa
     1681 taatgcacga aatcgttgct tatttttttt taaaagcggt atactagata taacgaaaca
     1741 acgaactgaa tagaaacgaa aaaagagcca tgacacattt ataaaatgtt tgacgacatt
     1801 ttataaatgc atagcccgat aagattgcca aaccaacgct tatcagttag tcagatgaac
     1861 tcttccctcg taagaagtta tttaattaac tttgtttgaa gacggtatat aaccgtacta
     1921 tcattatata gggaaatcag agagttttca agtatctaag ctactgaatt taagaattgt
     1981 taagcaatca atcggaaatc gtttgattgc tttttttgta ttcatttata gaaggtggag
     2041 tttgtatgaa tcatgatgaa tgtaaaactt atataaaaaa tagtttattg gagataagaa
     2101 aattagcaaa tatctataca ctagaaacgt ttaagaaaga gttagaaaag agaaatatct
     2161 acttagaaac aaaatcagat aagtattttt cttcggaggg ggaagattat atatataagt
     2221 taatagaaaa taacaaaata atttattcga ttagtggaaa aaaattgact tataaaggaa
     2281 aaaaatcttt ttcaaaacat gcaatattga aacagttgaa tgaaaaagca aaccaagtta
     2341 attaaacaac ctattttata ggatttatag gaaaggagaa cagctgaatg aatatccctt
     2401 ttgttgtaga aactgtgctt catgacggct tgttaaagta caaatttaaa aatagtaaaa
     2461 ttcgctcaat cactaccaag ccaggtaaaa gcaaaggggc tatttttgcg tatcgctcaa
     2521 aatcaagcat gattggcggt cgtggtgttg ttctgacttc cgaggaagcg attcaagaaa
     2581 atcaagatac atttacacat tggacaccca acgtttatcg ttatggaacg tatgcagacg
     2641 aaaaccgttc atacacgaaa ggacattctg aaaacaattt aagacaaatc aataccttct
     2701 ttattgattt tgatattcac acggcaaaag aaactatttc agcaagcgat attttaacaa
     2761 ccgctattga tttaggtttt atgcctacta tgattatcaa atctgataaa ggttatcaag
     2821 catattttgt tttagaaacg ccagtctatg tgacttcaaa atcagaattt aaatctgtca
     2881 aagcagccaa aataatttcg caaaatatcc gagaatattt tggaaagtct ttgccagttg
     2941 atctaacgtg taatcatttt ggtattgctc gcataccaag aacggacaat gtagaatttt
     3001 ttgatcctaa ttaccgttat tctttcaaag aatggcaaga ttggtctttc aaacaaacag
     3061 ataataaggg ctttactcgt tcaagtctaa cggttttaag cggtacagaa ggcaaaaaac
     3121 aagtagatga accctggttt aatctcttat tgcacgaaac gaaattttca ggagaaaagg
     3181 gtttaatagg gcgtaataac gtcatgttta ccctctcttt agcctacttt agttcaggct
     3241 attcaatcga aacgtgcgaa tataatatgt ttgagtttaa taatcgatta gatcaaccct
     3301 tagaagaaaa agaagtaatc aaaattgtta gaagtgccta ttcagaaaac tatcaagggg
     3361 ctaataggga atacattacc attctttgca aagcttgggt atcaagtgat ttaaccagta
     3421 aagatttatt tgtccgtcaa gggtggttta aattcaagaa aaaaagaagc gaacgtcaac
     3481 gtgttcattt gtcagaatgg aaagaagatt taatggctta tattagcgaa aaaagcgatg
     3541 tatacaagcc ttatttagtg acgaccaaaa aagagattag agaagtgcta ggcattcctg
     3601 aacggacatt agataaattg ctgaaggtac tgaaggcgaa tcaggaaatt ttctttaaga
     3661 ttaaaccagg aagaaatggt ggcattcaac ttgctagtgt taaatcattg ttgctatcga
     3721 tcattaaagt aaaaaaagaa gaaaaagaaa gctatataaa ggcgctgaca aattcttttg
     3781 acttagagca tacattcatt caagagactt taaacaagct agcagaacgc cctaaaacgg
     3841 acacacaact cgatttgttt agctatgata caggctgaaa ataaaacccg cactatgcca
     3901 ttacatttat atctatgata cgtgtttgtt ttttctttgc tgtttagcga atgattagca
     3961 gaaatataca gagtaagatt ttaattaatt attaggggga gaaggagaga gtagcccgaa
     4021 aacttttagt tggcttggac tgaacgaagt gagggaaagg ctactaaaac gtcgaggggc
     4081 agtgagagcg aagcgaacac ttgatttttt aattttctat cttttatagg tcattagagt
     4141 atacttattt gtcctataaa ctatttagca gcataataga tttattgaat aggtcattta
     4201 agttgagcat attagaggag gaaaatcttg gagaaatatt tgaagaaccc gattacatgg
     4261 attggattag ttcttgtggt tacgtggttt ttaactaaaa gtagtgaatt tttgattttt
     4321 ggtgtgtgtg tcttgttgtt agtatttgct agtcaaagtg attaaataga attctcatgt
     4381 ttgacagctt atcatcggag ctccgatgat aagctgtcca aacatgagaa ttccccggct
     4441 ttaggtatag tgtgtatctc aatccttggt atattgaaaa gaaagactaa aaattgatag
     4501 attatatttc ttcagaatga atggtataat gaagtaatga gtactaaaca atcggaggta
     4561 aagtggtgta taaaatttta atagttgatg atgatcagga aattttaaaa ttaatgaaga
     4621 cagcattaga aatgagaaac tatgaagttg cgacgcatca aaacatttca cttcccttgg
     4681 atattactga ttttcaggga tttgatttga ttttgttaga tatcatgatg tcaaatattg
     4741 aagggacaga aatttgtaaa aggattcgca gagaaatatc aactccaatt atctttgtta
     4801 gtgcgaaaga tacagaagag gatattataa acggcttagg tattggtggg gatgactata
     4861 ttactaagcc ttttagcctt aaacagttgg ttgcaaaagt ggaagcaaat ataaagcgag
     4921 aggaacgcaa taaacatgca gttcatgttt tttcagagat tcgtagagat ttaggaccaa
     4981 ttacatttta tttagaagaa aggcgagtct gtgtcaatgg tcaaacaatt ccactgactt
     5041 gtcgtgaata cgatattctt gaattactat cacaacgaac ttctaaagtt tatacgagag
     5101 aggatattta tgatgacgta tatgatgaat attctaatgc actttttcgg tcaatctcgg
     5161 agtatattta tcagattagg agtaagtttg caccatacga tattaatccg ataaaaacgg
     5221 ttcggggact tgggtatcag tggcatgggt aaaaaatatt caatgcgtcg acggatatgg
     5281 caagctgtca ttgaaattat cataggtact tgtctactta tcctgttgtt actgggcttg
     5341 actttctttc tacgacaaat tggacaaatc agtggttcag aaactattcg tttatcttta
     5401 gattcagata atttaactat ttctgatatc gaacgtgata tgaaacacta cccatatgat
     5461 tatattattt ttgacaatga tacaagtaaa attttgggag gacattatgt caagtcggat
     5521 gtacctagtt ttgtagcttc aaaacagtct tcacataata ttacagaagg agaaattact
     5581 tatacttatt caagcaataa gcatttttca gttgttttaa gacaaaacag tatgcctgaa
     5641 tttacaaatc atacgcttcg ttcaatttct tataatcaat ttacttacct tttctttttt
     5701 cttggtgaaa taatactcat tattttttct gtctatcatc tcattagaga attttctaag
     5761 aattttcaag ccgttcaaaa gattgcattg aagatggggg aaataactac ttttcctgaa
     5821 caagaggaat caaaaattat tgaatttgat caggttctga ataacttata ttcgaaaagt
     5881 aaggagttag ctttccttat tgaagcggag cgtcatgaaa aacatgattt atccttccag
     5941 gttgctgcac tttcacatga tgttaagaca cctttaacag tattaaaagg aaatattgaa
     6001 ctgctagaga tgactgaagt aaatgaacaa caagctgatt ttattgagtc aatgaaaaat
     6061 agtttgactg tttttgacaa gtattttaac acaatgatta gttatacaaa acttttgaat
     6121 gatgaaaatg attacaaagc gacaatctcc ctggaggatt ttttgataga tttatcagtt
     6181 gagttggaag agttgtcaac aacttatcaa gtggattatc agctagttaa aaaaacagat
     6241 ttaaccactt tttacggaaa tacattagct ttaagtcgag cacttatcaa tatctttgtt
     6301 aatgcctgtc agtatgctaa agagggtgaa aaaatagtca gtttgagtat ttatgatgat
     6361 gaaaaatatc tctattttga aatctggaat aatggtcatc ctttttctga acaagcaaaa
     6421 aaaaatgctg gaaaactatt tttcacagaa gatactggac gtagtgggaa acactatggg
     6481 attggactat cttttgctca aggtgtagct ttaaaacatc aaggaaactt aattctcagt
     6541 aatcctcaaa aaggtggggc agaagttatc ctaaaaataa aaaagtaatt tagtaatctc
     6601 taaggattac tttttttgtt tcggcaaact attaactggc gaactactta ctctagcttc
     6661 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6721 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6781 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6841 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6901 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     6961 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     7021 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7081 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7141 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7201 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     7261 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7321 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7381 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7441 gcgaacgacc tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct
     7501 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7561 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7621 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7681 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     7741 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     7801 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     7861 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcaggttaac
     7921 ctggcttatc gaaattaata cgactcacta tagggagacc ggcctcgagt ctagaagcgc
     7981 tgtcgacgag ctcgcatgcg aattcctgca gcccgggact agtggtaccg gatccatgca
     8041 gagtctcctg ttttacaacc gggtgtacat agcgaaatac ttgtaatgcg tggtgatgca
     8101 cctgaatctt tcttcgaaac agataccaaa tccaagctaa aatcttttgt actcattttg
     8161 agtgcctcct tataatttat tttgtagttc cttcgaacga aatcattgta tctaacaaac
     8221 ttcagaattt aatcagagcc gtttattatg ctcgcgttat cgacaataat attattacca
     8281 atactttctc aagatagaat taagactgtt ttagatttgt taatgtttct attgtcagta
     8341 tagttataag act
//
