LOCUS       V004455                 9463 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004455
VERSION     V004455
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9463)
  AUTHORS   Oddone GM, Mills DA, Block DE.
  TITLE     Incorporation of nisI-mediated nisin immunity improves vector-based
            nisin-controlled gene expression in lactic acid bacteria
  JOURNAL   Plasmid 61 (3), 151-158 (2009)
   PUBMED   19141301
REFERENCE   2  (bases 1 to 9463)
  AUTHORS   Oddone GM, Kim J-H.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-DEC-2008) Viticulture and Enology, University of
            California, Davis, RMI North Lab, 595 Hilgard Lane, Davis, CA 95616,
            USA
REFERENCE   3  (bases 1 to 9463)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9463)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid";
            date: "2009"; volume: "61"; issue: "3"; pages: "151-158"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (16-DEC-2008) Viticulture and Enology, University of California,
            Davis, RMI North Lab, 595 Hilgard Lane, Davis, CA 95616, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9463
                     /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             1971..2270
                     /codon_start=1
                     /gene="repD"
                     /product="RepD"
                     /label="repD"
                     /note="replication factor"
                     /protein_id="ACM62692.1"
                     /translation="MNHDECKTYIKNSLLEIRKLANIYTLETFKKELEKRNIYLETKSD
                     KYFSSEGEDYIYKLIENNKIIYSISGKKLTYKGKKSFSKHAILKQLNEKANQVN"
     gene            1971..2270
                     /gene="repD"
                     /label="repD"
     CDS             2313..3803
                     /codon_start=1
                     /gene="repE"
                     /product="RepE"
                     /label="repE"
                     /note="replication factor"
                     /protein_id="ACM62693.1"
                     /translation="MNIPFVVETVLHDGLLKYKFKNSKIRSITTKPGKSKGAIFAYRSK
                     SSMIGGRGVVLTSEEAIQENQDTFTHWTPNVYRYGTYADENRSYTKGHSENNLRQINTF
                     FIDFDIHTAKETISASDILTTAIDLGFMPTMIIKSDKGYQAYFVLETPVYVTSKSEFKS
                     VKAAKIISQNIREYFGKSLPVDLTCNHFGIARIPRTDNVEFFDPNYRYSFKEWQDWSFK
                     QTDNKGFTRSSLTVLSGTEGKKQVDEPWFNLLLHETKFSGEKGLIGRNNVMFTLSLAYF
                     SSGYSIETCEYNMFEFNNRLDQPLEEKEVIKIVRSAYSENYQGANREYITILCKAWVSS
                     DLTSKDLFVRQGWFKFKKKRSERQRVHLSEWKEDLMAYISEKSDVYKPYLVTTKKEIRE
                     VLGIPERTLDKLLKVLKANQEIFFKIKPGRNGGIQLASVKSLLLSIIKVKKEEKESYIK
                     ALTNSFDLEHTFIQETLNKLAERPKTDTQLDLFSYDTG"
     gene            2313..3803
                     /gene="repE"
                     /label="repE"
     CDS             4153..4290
                     /codon_start=1
                     /gene="repG"
                     /product="RepG"
                     /label="repG"
                     /note="replication factor"
                     /protein_id="ACM62694.1"
                     /translation="MEKYLKNPITWIGLVLVVTWFLTKSSEFLIFGVCVLLLVFASQSD
                     "
     gene            4153..4290
                     /gene="repG"
                     /label="repG"
     CDS             4440..5177
                     /codon_start=1
                     /gene="nisI"
                     /product="NisI"
                     /label="nisI"
                     /note="nisin immunity protein; provides significant
                     protection from the toxic effects of the inducer nisin"
                     /protein_id="ACM62695.1"
                     /translation="MRKYLILIVALIGITGLSGCYQTSQKKVRFDEGSYTNFIYDNKSY
                     FVTDKEIPQENVNNCKVKFYNLLIVDMKSEKLLSSSNKNSVTLVLNNIYEASDKSLCMG
                     INDRYYKILPESDKGVVKALRLQNFDVTSDISDDNFVIDKNDSRKIDYMGNIYSISDTT
                     VSDEELGEYQDVLAEVRVFDSVSGKSIPRSEWGRIDKDGSNSKQSRTEWDYGEIHSIRG
                     KSLTEAFAVEINDDFKLATKVGN"
     gene            4440..5177
                     /gene="nisI"
                     /label="nisI"
     CDS             5406..6044
                     /codon_start=1
                     /gene="nisR"
                     /product="NisR"
                     /label="nisR"
                     /note="nis operon regulator protein"
                     /protein_id="ACM62696.1"
                     /translation="MKTALEMRNYEVATHQNISLPLDITDFQGFDLILLDIMMSNIEGT
                     EICKRIRREISTPIIFVSAKDTEEDIINGLGIGGDDYITKPFSLKQLVAKVEANIKREE
                     RNKHAVHVFSEIRRDLGPITFYLEERRVCVNGQTIPLTCREYDILELLSQRTSKVYTRE
                     DIYDDVYDEYSNALFRSISEYIYQIRSKFAPYDINPIKTVRGLGYQWHG"
     gene            5406..6044
                     /gene="nisR"
                     /label="nisR"
     CDS             6037..7377
                     /gene="nisK"
                     /label="Nisin biosynthesis sensor protein NisK"
                     /note="Nisin biosynthesis sensor protein NisK from
                     Lactococcus lactis subsp. lactis. Accession#: P42707"
     rep_origin      7882..8470
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     promoter        8728..8746
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     9147..9163
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    9164..9220
                     /label="MCS"
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(9233..9249)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
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 gctcttatca tggggaagct
     1201 cggatcatat gcaagacaaa ataaactcgc aacagcactt ggagaaatgg gacgaatcga
     1261 gaaaaccctc tttacgctgg attacatatc taataaagcc gtaaggagac gggttcaaaa
     1321 aggtttaaat aaaggagaag caatcaatgc atccggacgc gtgccggcga cgtcgtacgg
     1381 taccccaaga attagaaatg agtagatcaa attattcacg aatagaatca ggaaaatcag
     1441 atccaaccat aaaaacacta gaacaaattg caaagttaac taactcaacg ctagtagtgg
     1501 atttaatccc aaatgagcca acagaaccag agccagaaac agaatcagaa caagtaacat
     1561 tggatttaga aatggaagaa gaaaaaagca atgacttcgt gtgaataatg cacgaaatcg
     1621 ttgcttattt ttttttaaaa gcggtatact agatataacg aaacaacgaa ctgaatagaa
     1681 acgaaaaaag agccatgaca catttataaa atgtttgacg acattttata actgcatagc
     1741 ccgataagat tgccaaacca acgcttatca gttagtcaga tgaactcttc cctcgtaaga
     1801 agttatttaa ttaactttgt ttgaagacgg tatataaccg tactatcatt atatagggaa
     1861 atcagagagt tttcaagtat ctaagctact gaatttaaga attgttaagc aatcaatcgg
     1921 aaatcgtttg attgcttttt ttgtattcat ttatagaagg tggagtttgt atgaatcatg
     1981 atgaatgtaa aacttatata aaaaatagtt tattggagat aagaaaatta gcaaatatct
     2041 atacactaga aacgtttaag aaagagttag aaaagagaaa tatctactta gaaacaaaat
     2101 cagataagta tttttcttcg gagggggaag attatatata taagttaata gaaaataaca
     2161 aaataattta ttcgattagt ggaaaaaaat tgacttataa aggaaaaaaa tctttttcaa
     2221 aacatgcaat attgaaacag ttgaatgaaa aagcaaacca agttaattaa acaacctatt
     2281 ttataggatt tataggaaag gagaacagct gaatgaatat cccttttgtt gtagaaactg
     2341 tgcttcatga cggcttgtta aagtacaaat ttaaaaatag taaaattcgc tcaatcacta
     2401 ccaagccagg taaaagcaaa ggggctattt ttgcgtatcg ctcaaaatca agcatgattg
     2461 gcggtcgtgg tgttgttctg acttccgagg aagcgattca agaaaatcaa gatacattta
     2521 cacattggac acccaacgtt tatcgttatg gaacgtatgc agacgaaaac cgttcataca
     2581 cgaaaggaca ttctgaaaac aatttaagac aaatcaatac cttctttatt gattttgata
     2641 ttcacacggc aaaagaaact atttcagcaa gcgatatttt aacaaccgct attgatttag
     2701 gttttatgcc tactatgatt atcaaatctg ataaaggtta tcaagcatat tttgttttag
     2761 aaacgccagt ctatgtgact tcaaaatcag aatttaaatc tgtcaaagca gccaaaataa
     2821 tttcgcaaaa tatccgagaa tattttggaa agtctttgcc agttgatcta acgtgtaatc
     2881 attttggtat tgctcgcata ccaagaacgg acaatgtaga attttttgat cctaattacc
     2941 gttattcttt caaagaatgg caagattggt ctttcaaaca aacagataat aagggcttta
     3001 ctcgttcaag tctaacggtt ttaagcggta cagaaggcaa aaaacaagta gatgaaccct
     3061 ggtttaatct cttattgcac gaaacgaaat tttcaggaga aaagggttta atagggcgta
     3121 ataacgtcat gtttaccctc tctttagcct actttagttc aggctattca atcgaaacgt
     3181 gcgaatataa tatgtttgag tttaataatc gattagatca acccttagaa gaaaaagaag
     3241 taatcaaaat tgttagaagt gcctattcag aaaactatca aggggctaat agggaataca
     3301 ttaccattct ttgcaaagct tgggtatcaa gtgatttaac cagtaaagat ttatttgtcc
     3361 gtcaagggtg gtttaaattc aagaaaaaaa gaagcgaacg tcaacgtgtt catttgtcag
     3421 aatggaaaga agatttaatg gcttatatta gcgaaaaaag cgatgtatac aagccttatt
     3481 tagtgacgac caaaaaagag attagagaag tgctaggcat tcctgaacgg acattagata
     3541 aattgctgaa ggtactgaag gcgaatcagg aaattttctt taagattaaa ccaggaagaa
     3601 atggtggcat tcaacttgct agtgttaaat cattgttgct atcgatcatt aaagtaaaaa
     3661 aagaagaaaa agaaagctat ataaaggcgc tgacaaattc ttttgactta gagcatacat
     3721 tcattcaaga gactttaaac aagctagcag aacgccctaa aacggacaca caactcgatt
     3781 tgtttagcta tgatacaggc tgaaaataaa acccgcacta tgccattaca tttatatcta
     3841 tgatacgtgt ttgttttttc tttgctgttt agcgaatgat tagcagaaat atacagagta
     3901 agattttaat taattattag ggggagaagg agagagtagc ccgaaaactt ttagttggct
     3961 tggactgaac gaagtgaggg aaaggctact aaaacgtcga ggggcagtga gagcgaagcg
     4021 aacacttgat tttttaattt tctatctttt ataggtcatt agagtatact tatttgtcct
     4081 ataaactatt tagcagcata atagatttat tgaataggtc atttaagttg agcatattag
     4141 aggaggaaaa tcttggagaa atatttgaag aacccgatta catggattgg attagttctt
     4201 gtggttacgt ggtttttaac taaaagtagt gaatttttga tttttggtgt gtgtgtcttg
     4261 ttgttagtat ttgctagtca aagtgattaa atagaattcg atgaaagtgg cacgggtttt
     4321 cttgaaggaa taagtggctg tatactggta ttatcgaaat ttgaatattc aatcaatttt
     4381 acttattgga gacaagcact gttacttttt gatgattttt tgaaaggagg gaagaggaaa
     4441 tgagaaaata tttaatactt attgtggcct taatagggat aacaggttta tcagggtgtt
     4501 atcaaacaag tcaaaaaaag gtgaggtttg acgaaggaag ttatactaat tttatttatg
     4561 ataataaatc gtatttcgta actgataagg agattcctca ggagaacgtt aacaattgca
     4621 aagtaaaatt ttataacctg ttgattgttg acatgaaaag tgagaaactt ttatcaagta
     4681 gcaacaaaaa tagtgtgact ttggtcttaa ataatattta tgaggcttct gacaagtcgc
     4741 tatgtatggg tattaacgac agatactata agatacttcc agaaagtgat aagggggtgg
     4801 tcaaagcttt gagattacaa aactttgatg tgacaagcga tatttctgat gataattttg
     4861 ttattgataa aaatgattca cgaaaaattg actatatggg aaatatttac agtatatcgg
     4921 acaccaccgt atctgatgaa gaattgggag aatatcagga tgttttagct gaagtacgtg
     4981 tgtttgattc agttagtggc aaaagtatcc cgaggtctga atgggggaga attgataagg
     5041 atggttcaaa ttctaaacag agtaggacgg aatgggatta tggcgaaatc cattctatta
     5101 gaggaaaatc tcttactgaa gcatttgccg ttgagataaa tgatgatttt aagcttgcaa
     5161 cgaaggtagg aaactagagt gaaaaaaata ctaggtttcc tttttatcgt ttgttcgttg
     5221 aattccccgg ctttaggtat agtgtgtatc tcaatccttg gtatattgaa aagaaagact
     5281 aaaaattgat agattatatt tcttcagaat gaatggtata atgaagtaat gagtactaaa
     5341 caatcggagg taaagtggtg tataaaattt taatagttga tgatgatcag gaaattttaa
     5401 aattaatgaa gacagcatta gaaatgagaa actatgaagt tgcgacgcat caaaacattt
     5461 cacttccctt ggatattact gattttcagg gatttgattt gattttgtta gatatcatga
     5521 tgtcaaatat tgaagggaca gaaatttgta aaaggattcg cagagaaata tcaactccaa
     5581 ttatctttgt tagtgcgaaa gatacagaag aggatattat aaacggctta ggtattggtg
     5641 gggatgacta tattactaag ccttttagcc ttaaacagtt ggttgcaaaa gtggaagcaa
     5701 atataaagcg agaggaacgc aataaacatg cagttcatgt tttttcagag attcgtagag
     5761 atttaggacc aattacattt tatttagaag aaaggcgagt ctgtgtcaat ggtcaaacaa
     5821 ttccactgac ttgtcgtgaa tacgatattc ttgaattact atcacaacga acttctaaag
     5881 tttatacgag agaggatatt tatgatgacg tatatgatga atattctaat gcactttttc
     5941 ggtcaatctc ggagtatatt tatcagatta ggagtaagtt tgcaccatac gatattaatc
     6001 cgataaaaac ggttcgggga cttgggtatc agtggcatgg gtaaaaaata ttcaatgcgt
     6061 cgacggatat ggcaagctgt cattgaaatt atcataggta cttgtctact tatcctgttg
     6121 ttactgggct tgactttctt tctacgacaa attggacaaa tcagtggttc agaaactatt
     6181 cgtttatctt tagattcaga taatttaact atttctgata tcgaacgtga tatgaaacac
     6241 tacccatatg attatattat ttttgacaat gatacaagta aaattttggg aggacattat
     6301 gtcaagtcgg atgtacctag ttttgtagct tcaaaacagt cttcacataa tattacagaa
     6361 ggagaaatta cttatactta ttcaagcaat aagcattttt cagttgtttt aagacaaaac
     6421 agtatgcctg aatttacaaa tcatacgctt cgttcaattt cttataatca atttacttac
     6481 cttttctttt ttcttggtga aataatactc attatttttt ctgtctatca tctcattaga
     6541 gaattttcta agaattttca agccgttcaa aagattgcat tgaagatggg ggaaataact
     6601 acttttcctg aacaagagga atcaaaaatt attgaatttg atcaggttct gaataactta
     6661 tattcgaaaa gtaaggagtt agctttcctt attgaagcgg agcgtcatga aaaacatgat
     6721 ttatccttcc aggttgctgc actttcacat gatgttaaga cacctttaac agtattaaaa
     6781 ggaaatattg aactgctaga gatgactgaa gtaaatgaac aacaagctga ttttattgag
     6841 tcaatgaaaa atagtttgac tgtttttgac aagtatttta acacaatgat tagttataca
     6901 aaacttttga atgatgaaaa tgattacaaa gcgacaatct ccctggagga ttttttgata
     6961 gatttatcag ttgagttgga agagttgtca acaacttatc aagtggatta tcagctagtt
     7021 aaaaaaacag atttaaccac tttttacgga aatacattag ctttaagtcg agcacttatc
     7081 aatatctttg ttaatgcctg tcagtatgct aaagagggtg aaaaaatagt cagtttgagt
     7141 atttatgatg atgaaaaata tctctatttt gaaatctgga ataatggtca tcctttttct
     7201 gaacaagcaa aaaaaaatgc tggaaaacta tttttcacag aagatactgg acgtagtggg
     7261 aaacactatg ggattggact atcttttgct caaggtgtag ctttaaaaca tcaaggaaac
     7321 ttaattctca gtaatcctca aaaaggtggg gcagaagtta tcctaaaaat aaaaaagtaa
     7381 tttagtaatc tctaaggatt actttttttg tttcggcaaa ctattaactg gcgaactact
     7441 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     7501 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     7561 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     7621 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     7681 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     7741 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     7801 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7861 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7921 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7981 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     8041 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     8101 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     8161 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     8221 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga
     8281 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     8341 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     8401 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     8461 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     8521 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     8581 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     8641 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     8701 atgcaggtta acctggctta tcgaaattaa tacgactcac tatagggaga ccggcctcga
     8761 gatttacaaa aataaaaaag tctgttgata acagactttt ttattatact tttagcttaa
     8821 caaatttaat ttaaaataaa aagagctagt tgaaatctca accaactctt tttatttcct
     8881 aggaccaggt ccgcggccgg gcccgttatt tgtctatgcg gcatcagagc agattgtact
     8941 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     9001 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     9061 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     9121 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattcg agctcggtac
     9181 gcggggatcc tccagagtcg acctgcaggc atgcaagctt ggcgtaatca tggtcatagc
     9241 tgtttcctgt gtgaaattga ccggtcgcga cctggtctag aataatttat tttgtagttc
     9301 cttcgaacga aatcattgta tctaacaaac ttcagaattt aatcagagcc gtttattatg
     9361 ctcgcgttat cgacaataat attattacca atactttctc aagatagaat taagactgtt
     9421 ttagatttgt taatgtttct attgtcagta tagttataag act
//
