LOCUS       V004414                 6297 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004414
VERSION     V004414
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6297)
  AUTHORS   Heap JT, Pennington OJ, Cartman ST, Minton NP.
  TITLE     A modular system for Clostridium shuttle plasmids
  JOURNAL   J. Microbiol. Methods 78 (1), 79-85 (2009)
   PUBMED   19445976
REFERENCE   2  (bases 1 to 6297)
  AUTHORS   Pennington OJ, Heap JT, Cartman ST, Minton NP.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-MAR-2009) Centre for Biomolecular Sciences, The
            University of Nottingham, University Park, Nottingham NG7 2RD,
            United Kingdom
REFERENCE   3  (bases 1 to 6297)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6297)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J.
            Microbiol. Methods"; date: "2009"; volume: "78"; issue: "1"; pages:
            "79-85"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (02-MAR-2009) Centre for Biomolecular Sciences, The University of
            Nottingham, University Park, Nottingham NG7 2RD, United Kingdom"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      9..62
                     /note="terminator derived from CD0164 gene of Clostridium
                     difficile"
                     /regulatory_class="terminator"
     primer_bind     63..79
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     primer_bind     complement(199..215)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             569..1216
                     /codon_start=1
                     /product="putative plasmid replication protein"
                     /label="putative plasmid replication protein"
                     /note="orfB"
                     /protein_id="ACR43904.1"
                     /translation="MNKKLEKPFVYKREYDLTGYDVEILQKYELEQAIYVYVGSSCAYN
                     MRARSSKWRYHIRTNNKSICCNIKNFIHNLELFYKMELKLSDNIINDKLYYSNIAEFEE
                     FETLEKAREVESTIISQYQFLDSINHMLKQKIILLSNKDSVLNITKNGNTNYLKVKNKY
                     IEKHKNKPIMRYHINCQFNTDGSVKSITQEFEPILELNKKNTLSRPSRVFLK"
     CDS             1491..3128
                     /codon_start=1
                     /gene="repA"
                     /product="plasmid replication protein"
                     /label="repA"
                     /protein_id="ACR43903.1"
                     /translation="MEQLDSKYKLKKFLMAVFRDGIGQGNNLIDNEYVRVFQNNKSNSK
                     QLELGEEFKEYSKTTFFKNIDDIVEFTFAKNIYYENTFFNLCTTDGKAGTNENLINRYA
                     LGFDFDKKELGQGFNYKDIINLFTKIGLHYHILVDSGNGFHVYVLINKTNNIKLVSEVT
                     NTLINKLGADKQANLSTQVLRVPYTYNIKNTTKQVKIIHQDKNIYRYDIEKLAKKYCKD
                     VKTVGNTNTKYILDSKLPNCIVDILKNGSKDGHKNLDLQKIVVTLRLRNKSLSQVISVA
                     REWNYISQNSLSNSELEYQVKYMYEKLKTVNFGCTGCEFNSDCWNKIESDFIYSDEDTL
                     FNMPHKHSKDLKYKNRKGVKIMTGNQLFIYNVLLNNKDRELNIDDIMELITYKRKKKVK
                     NIVMSEKTLRETLKELQHNDYITKTKGVTKLGIKDTYNVKEVRCNIDKQYTISYFVTMA
                     VIWGIISTEELRLYTHMRYKQDLLVKDDKIKGNILRINQEELAKDLGVTQQRISNMIES
                     LLDTKILDVWETKINDRGFMYYTYRLNK"
     gene            1491..3128
                     /gene="repA"
                     /label="repA"
     CDS             3941..4561
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      4741..5329
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     oriT            5650..5759
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             5792..6160
                     /label="traJ"
                     /note="oriT-recognizing protein"
ORIGIN
        1 cctgcaggat aaaaaaattg tagataaatt ttataaaata gttttatcta caattttttt
       61 atcaggaaac agctatgacc gcggccgctg tatccatatg accatgatta cgaattcgag
      121 ctcggtaccc ggggatcctc tagagtcgac gtcacgcgtc catggagatc tcgaggcctg
      181 cagacatgca agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
      241 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
      301 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcta
      361 gcataaaaat aagaagcctg catttgcagg cttcttattt ttatggcgcg cccgccctta
      421 agtctaaaaa ttaggggaga tgtaaggatt tgggaaaaat agaagatgtt ataatcataa
      481 atatggtatt cgtaggctta aagtcaaaaa ggaggtgaaa tataaataga tttttagcta
      541 aattaagtaa gaaataggag gagatttatt gaacaaaaaa ttagaaaaac catttgtata
      601 taagagagag tacgatttga ctggatatga tgttgaaatt ttacaaaaat atgagttaga
      661 acaagcaata tatgtttatg ttgggagtag ttgtgcatat aacatgagag ctagaagtag
      721 taaatggaga taccatataa gaacaaataa taagtctata tgttgtaaca ttaaaaattt
      781 tatacataac ttggaattgt tttataaaat ggaattaaag ttgtcagata atattattaa
      841 tgataagcta tactatagca atatagcaga gtttgaagaa tttgaaacac tagaaaaagc
      901 tagagaggta gaaagtacta ttataagtca atatcaattt ttagattcta taaatcacat
      961 gttaaaacaa aaaataattt tattgagtaa taaggatagt gtgttaaaca taactaaaaa
     1021 tggaaataca aattatttga aagtaaaaaa taaatacata gaaaaacata agaacaagcc
     1081 aataatgaga taccatatca actgtcaatt caatacagat ggaagtgtca aaagtattac
     1141 acaggagttt gaaccaatat tggaattaaa caaaaaaaat accctaagcc gaccaagcag
     1201 agtattttta aaataatatt ttaagataac aacaaaatga gataatacta ctagacaatg
     1261 acaactcaac taccaattga gtttatggag ctaccaactc caatatcggt ctaactgatt
     1321 aagtatctgt agttatataa taatattgct atcaatttta gcatcttaac aatattatta
     1381 tacatactaa gctaaaatta ttcaatagtt gtaaaagttg attagtcaat aagtatatat
     1441 ttaatgtagt gttatctctt aaaaaaacta gataaggaga taataaatat atggaacaat
     1501 tagattcaaa atataagttg aaaaaatttc taatggcagt atttagagat ggtataggac
     1561 aaggaaataa tcttattgat aatgaatatg ttagagtatt tcaaaataat aaaagtaata
     1621 gtaaacaatt agaactcgga gaagaattta aagaatatag taaaacaact ttttttaaaa
     1681 atatagatga tatagtagaa tttaccttcg caaaaaatat ttattatgaa aatacatttt
     1741 ttaacctatg tactactgat ggaaaagcag gaaccaatga aaacttaata aatagatatg
     1801 cattaggatt tgattttgac aaaaaagaat taggacaagg ttttaattat aaagatataa
     1861 ttaatttatt tactaagata ggattacatt atcatatcct agttgatagt ggaaatggat
     1921 tccatgttta tgtgctaatt aataaaacta ataacattaa gttagtatca gaagttacaa
     1981 atacattaat aaataaattg ggtgcagata aacaagcaaa tttatctact caagtattaa
     2041 gagtacctta tacatataat attaaaaata ctactaaaca agtaaaaata atacaccaag
     2101 acaaaaatat atatagatat gacatagaaa agttagctaa aaaatattgc aaagatgtaa
     2161 aaacagtagg taatactaat acaaaatata tattagatag taagctacca aattgtatag
     2221 tagatatttt aaaaaatggt agtaaagatg gacataaaaa cctagatttg caaaaaatag
     2281 ttgtgacttt aagattgagg aataaaagtt taagtcaagt aatatccgtt gctagagaat
     2341 ggaactatat atcacaaaat agtctttcaa atagtgagct agaatatcaa gtcaagtata
     2401 tgtatgagaa acttaaaacg gttaattttg gttgtactgg ttgtgagttt aatagtgatt
     2461 gttggaataa aatagaatca gattttatat atagtgatga agatactttg ttcaatatgc
     2521 cacataagca ctcaaaggat ttgaaatata agaataggaa aggggttaaa ataatgactg
     2581 gtaatcaatt gtttatctat aatgtgttac ttaacaataa agatagagaa ttaaacatag
     2641 acgatataat ggagctgata acctataaac gtaagaagaa agttaaaaac attgttatga
     2701 gtgaaaagac attaagagaa acattaaaag aacttcaaca taatgattat attacaaaaa
     2761 caaaaggtgt tacaaagcta ggaataaaag atacatacaa tgtaaaagaa gttagatgta
     2821 atatagataa acaatatact attagttact ttgttaccat ggcagtaatt tggggaataa
     2881 tttcaactga agaattaaga ttatatactc acatgagata taagcaagat ttattggtca
     2941 aagatgataa aataaaagga aatatattaa gaattaatca agaggaatta gcaaaagatt
     3001 taggagtaac acagcaaaga atttcaaata tgatagaatc tttattagat actaaaattt
     3061 tagatgtatg ggaaactaaa ataaatgata gaggatttat gtactataca tatagattaa
     3121 acaagtagat ttttgatagg attagaattg attttctagt cctattttta tgcaaaaaaa
     3181 ctaattaata aaaatttctt ttggtaaaat aattgtacga gaattgcaaa aaaaaaatgg
     3241 catcaaagta ttgaaattaa gccgttttaa aaatttcttt tggtaaaata attctacata
     3301 tatatgtagt atatatatat atgtttttta gagaatgtat aactagaata tagagctaga
     3361 atatagagaa tgtataacta gaatatagag ctagaatata gagaatgtat aactagaata
     3421 tagagctaga atatagagaa tgtataacta gaatatagag ctagaatata gagaatgtat
     3481 aactagaata tagagctaga atatagagaa tgtataacta gaatatagag ctagaatata
     3541 gagaatgtat aactagaata tagagctaga atcctaatta gtaggtgctt ttttaaaaca
     3601 agttaaaaat caaaaatagt attagtaagc attggaaatg ctagattcta aaatagaaaa
     3661 gtaaaaaatt ggtgcactat ctaaacttat ctatatcgct ttttccgtcg tttggttctc
     3721 tagttacgat acaggggata tgcttatatt gagttatagt actaatcagt gcttaatata
     3781 gttaataaaa ttatagttac catagtttag taactatgat gtatgttagt tagaaacttg
     3841 catttcggcc ggccagtggg caagttgaaa aattcacaaa aatgtggtat aatatctttg
     3901 ttcattagag cgataaactt gaatttgaga gggaacttag atggtatttg aaaaaattga
     3961 taaaaatagt tggaacagaa aagagtattt tgaccactac tttgcaagtg taccttgtac
     4021 ctacagcatg accgttaaag tggatatcac acaaataaag gaaaagggaa tgaaactata
     4081 tcctgcaatg ctttattata ttgcaatgat tgtaaaccgc cattcagagt ttaggacggc
     4141 aatcaatcaa gatggtgaat tggggatata tgatgagatg ataccaagct atacaatatt
     4201 tcacaatgat actgaaacat tttccagcct ttggactgag tgtaagtctg actttaaatc
     4261 atttttagca gattatgaaa gtgatacgca acggtatgga aacaatcata gaatggaagg
     4321 aaagccaaat gctccggaaa acatttttaa tgtatctatg ataccgtggt caaccttcga
     4381 tggctttaat ctgaatttgc agaaaggata tgattatttg attcctattt ttactatggg
     4441 gaaatattat aaagaagata acaaaattat acttcctttg gcaattcaag ttcatcacgc
     4501 agtatgtgac ggatttcaca tttgccgttt tgtaaacgaa ttgcaggaat tgataaatag
     4561 ttaacttcag gtttgtctgt aactaaaaac aagtatttaa gcaaaaacat cgtagaaata
     4621 cggtgttttt tgttacccta agtttaaact cctttttgat aatctcatga ccaaaatccc
     4681 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4741 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4801 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4861 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4921 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4981 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5041 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5101 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5161 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5221 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5281 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5341 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5401 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     5461 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     5521 acgcagggcc ccctgcttcg gggtcattat agcgattttt tcggtatatc catccttttt
     5581 cgcacgatat acaggatttt gccaaagggt tcgtgtagac tttccttggt gtatccaacg
     5641 gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt ccttcttcac
     5701 tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag gctggccggc
     5761 taccgccggc gtaacagatg agggcaagcg gatggctgat gaaaccaagc caaccaggaa
     5821 gggcagccca cctatcaagg tgtactgcct tccagacgaa cgaagagcga ttgaggaaaa
     5881 ggcggcggcg gccggcatga gcctgtcggc ctacctgctg gccgtcggcc agggctacaa
     5941 aatcacgggc gtcgtggact atgagcacgt ccgcgagctg gcccgcatca atggcgacct
     6001 gggccgcctg ggcggcctgc tgaaactctg gctcaccgac gacccgcgca cggcgcggtt
     6061 cggtgatgcc acgatcctcg ccctgctggc gaagatcgaa gagaagcagg acgagcttgg
     6121 caaggtcatg atgggcgtgg tccgcccgag ggcagagcca tgactttttt agccgctaaa
     6181 acggccgggg ggtgcgcgtg attgccaagc acgtccccat gcgctccatc aagaagagcg
     6241 acttcgcgga gctggtgaag tacatcaccg acgagcaagg caagaccgat cgggccc
//
