LOCUS       V004433                 4230 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004433
VERSION     V004433
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4230)
  AUTHORS   Heap JT, Ehsaan M, Cooksley CM, Ng YK, Cartman ST, Winzer K, Minton
            NP.
  TITLE     Integration of DNA into bacterial chromosomes from plasmids without
            a counter-selection marker
  JOURNAL   Nucleic Acids Res. 40 (8), E59 (2012)
   PUBMED   22259038
REFERENCE   2  (bases 1 to 4230)
  AUTHORS   Heap JT, Ehsaan M, Cooksley CM, Cartman ST, Minton NP.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-JAN-2011) School of Molecular Medical Sciences, The
            University of Nottingham, Centre for Biomolecular Sciences,
            University Park, Nottingham, Nottinghamshire NG7 2RD, United Kingdom
REFERENCE   3  (bases 1 to 4230)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4230)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
            Acids Res."; date: "2012"; volume: "40"; issue: "8"; pages: "E59"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (11-JAN-2011) School of Molecular Medical Sciences, The University
            of Nottingham, Centre for Biomolecular Sciences, University Park,
            Nottingham, Nottinghamshire NG7 2RD, United Kingdom"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            9..581
                     /gene="pyrE fragment"
                     /label="pyrE fragment"
     misc_feature    9..308
                     /gene="pyrE fragment"
                     /note="mediates homologous recombination with the
                     chromosome of Clostridium sporogenes NCIMB10696"
     primer_bind     complement(700..716)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     regulatory      864..905
                     /label="derived from fdx gene of Clostridium pasteurianum"
                     /note="derived from fdx gene of Clostridium pasteurianum"
                     /regulatory_class="terminator"
     CDS             1284..1724
                     /codon_start=1
                     /gene="repL"
                     /product="plasmid replication protein"
                     /label="repL"
                     /protein_id="AFA34984.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     gene            1284..1724
                     /gene="repL"
                     /label="repL"
     CDS             1874..2494
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      2674..3262
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     oriT            3583..3692
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             3725..4093
                     /label="traJ"
                     /note="oriT-recognizing protein"
ORIGIN
        1 cctgcaggag taatataaat gttatagata tattaaaaga atcaaatgca ttattagaag
       61 gtcatttctt actatcatct ggaagacata gtaatagata ttgtcaatgt gcgaaactac
      121 ttcaatatcc acaaaaagca gaaaaagtta ttagtgtaat agcacagaag cttaaaaatg
      181 tagactttaa tataatagta ggaccagcta tgggtggtgt aatagtatcc tatgaattag
      241 caaggcaaac taatagacca ggcatatttg cagaaagaaa agatggaatc atgtgtatac
      301 gaagaggatt tgaaataaaa aaaggagata aagttataat atcagaggat gtaataacta
      361 caggtaaatc ttctttagaa gtagctaaag ttatagaaga aatgggtgga gaagtagtag
      421 gtatagcatg tatagttgat agaagagcgg aaggtataaa aacaaactat caaatatata
      481 gtgcttgtaa attagaaata gaaacttatg aaaaagataa ctgtgagtta tgtaaaaaaa
      541 atataccttt tgtaaaacca ggtagtagag aacaaaaata agcggccgct gtatccatat
      601 gaccatgatt acgaattcga gctcggtacc cggggatcct ctagagtcga cgtcacgcgt
      661 ccatggagat ctcgaggcct gcagacatgc aagcttggca ctggccgtcg ttttacaacg
      721 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
      781 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
      841 cctgaatggc gaatggcgct agcataaaaa taagaagcct gcatttgcag gcttcttatt
      901 tttatggcgc gccgcattca cttcttttct atataaatat gagcgaagcg aataagcgtc
      961 ggaaaagcag caaaaagttt cctttttgct gttggagcat gggggttcag ggggtgcagt
     1021 atctgacgtc aatgccgagc gaaagcgagc cgaagggtag catttacgtt agataacccc
     1081 ctgatatgct ccgacgcttt atatagaaaa gaagattcaa ctaggtaaaa tcttaatata
     1141 ggttgagatg ataaggttta taaggaattt gtttgttcta atttttcact cattttgttc
     1201 taatttcttt taacaaatgt tctttttttt ttagaacagt tatgatatag ttagaatagt
     1261 ttaaaataag gagtgagaaa aagatgaaag aaagatatgg aacagtctat aaaggctctc
     1321 agaggctcat agacgaagaa agtggagaag tcatagaggt agacaagtta taccgtaaac
     1381 aaacgtctgg taacttcgta aaggcatata tagtgcaatt aataagtatg ttagatatga
     1441 ttggcggaaa aaaacttaaa atcgttaact atatcctaga taatgtccac ttaagtaaca
     1501 atacaatgat agctacaaca agagaaatag caaaagctac aggaacaagt ctacaaacag
     1561 taataacaac acttaaaatc ttagaagaag gaaatattat aaaaagaaaa actggagtat
     1621 taatgttaaa ccctgaacta ctaatgagag gcgacgacca aaaacaaaaa tacctcttac
     1681 tcgaatttgg gaactttgag caagaggcaa atgaaataga ttgacctccc aataacacca
     1741 cgtagttatt gggaggtcaa tctatgaaat gcgattaagg gccggccagt gggcaagttg
     1801 aaaaattcac aaaaatgtgg tataatatct ttgttcatta gagcgataaa cttgaatttg
     1861 agagggaact tagatggtat ttgaaaaaat tgataaaaat agttggaaca gaaaagagta
     1921 ttttgaccac tactttgcaa gtgtaccttg tacctacagc atgaccgtta aagtggatat
     1981 cacacaaata aaggaaaagg gaatgaaact atatcctgca atgctttatt atattgcaat
     2041 gattgtaaac cgccattcag agtttaggac ggcaatcaat caagatggtg aattggggat
     2101 atatgatgag atgataccaa gctatacaat atttcacaat gatactgaaa cattttccag
     2161 cctttggact gagtgtaagt ctgactttaa atcattttta gcagattatg aaagtgatac
     2221 gcaacggtat ggaaacaatc atagaatgga aggaaagcca aatgctccgg aaaacatttt
     2281 taatgtatct atgataccgt ggtcaacctt cgatggcttt aatctgaatt tgcagaaagg
     2341 atatgattat ttgattccta tttttactat ggggaaatat tataaagaag ataacaaaat
     2401 tatacttcct ttggcaattc aagttcatca cgcagtatgt gacggatttc acatttgccg
     2461 ttttgtaaac gaattgcagg aattgataaa tagttaactt caggtttgtc tgtaactaaa
     2521 aacaagtatt taagcaaaaa catcgtagaa atacggtgtt ttttgttacc ctaagtttaa
     2581 actccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     2641 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2701 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2761 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2821 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2881 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2941 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3001 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3061 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3121 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3181 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3241 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3301 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3361 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3421 gtcagtgagc gaggaagcgg aagagcgccc aatacgcagg gccccctgct tcggggtcat
     3481 tatagcgatt ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag
     3541 ggttcgtgta gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt
     3601 aggcccaccc gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct
     3661 caacgggaat cctgctctgc gaggctggcc ggctaccgcc ggcgtaacag atgagggcaa
     3721 gcggatggct gatgaaacca agccaaccag gaagggcagc ccacctatca aggtgtactg
     3781 ccttccagac gaacgaagag cgattgagga aaaggcggcg gcggccggca tgagcctgtc
     3841 ggcctacctg ctggccgtcg gccagggcta caaaatcacg ggcgtcgtgg actatgagca
     3901 cgtccgcgag ctggcccgca tcaatggcga cctgggccgc ctgggcggcc tgctgaaact
     3961 ctggctcacc gacgacccgc gcacggcgcg gttcggtgat gccacgatcc tcgccctgct
     4021 ggcgaagatc gaagagaagc aggacgagct tggcaaggtc atgatgggcg tggtccgccc
     4081 gagggcagag ccatgacttt tttagccgct aaaacggccg gggggtgcgc gtgattgcca
     4141 agcacgtccc catgcgctcc atcaagaaga gcgacttcgc ggagctggtg aagtacatca
     4201 ccgacgagca aggcaagacc gatcgggccc
//
