LOCUS       V004446                 3191 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004446
VERSION     V004446
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 3191)
  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 3191)
  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 3191)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3191)
  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..3191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    9..308
                     /gene="pyrE fragment"
                     /note="mediates homologous recombination with the
                     chromosome of Clostridium acetobutylicum ATCC824"
     gene            9..308
                     /gene="pyrE fragment"
                     /label="pyrE fragment"
     primer_bind     complement(427..443)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     regulatory      591..632
                     /label="derived from fdx gene of Clostridium pasteurianum"
                     /note="derived from fdx gene of Clostridium pasteurianum"
                     /regulatory_class="terminator"
     CDS             1011..1451
                     /codon_start=1
                     /gene="repL"
                     /product="plasmid replication protein"
                     /label="repL"
                     /protein_id="AFA34936.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     gene            1011..1451
                     /gene="repL"
                     /label="repL"
     CDS             1601..2221
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      2401..2989
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 cctgcaggag agtaatgtac ttacctttgg ggatttcata actaaaagcg gcagaagaac
       61 accatttttt ataaatacag gtaactacaa gacaggtaat caattaaata agttggctaa
      121 gttttatgct aaagcaatat atgataattt tggagatgat tttgatattt tatttgggcc
      181 tgcatataaa ggaatacctt taagtgtttc agtagctatg gcacttgata atatttatgg
      241 aattaatgca gcttattgtt caaatagaaa agaagttaaa gatcacggtg ataagggaat
      301 acttcttggc ggccgctgta tccatatgac catgattacg aattcgagct cggtacccgg
      361 ggatcctcta gagtcgacgt cacgcgtcca tggagatctc gaggcctgca gacatgcaag
      421 cttggcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
      481 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
      541 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctagc ataaaaataa
      601 gaagcctgca tttgcaggct tcttattttt atggcgcgcc gcattcactt cttttctata
      661 taaatatgag cgaagcgaat aagcgtcgga aaagcagcaa aaagtttcct ttttgctgtt
      721 ggagcatggg ggttcagggg gtgcagtatc tgacgtcaat gccgagcgaa agcgagccga
      781 agggtagcat ttacgttaga taaccccctg atatgctccg acgctttata tagaaaagaa
      841 gattcaacta ggtaaaatct taatataggt tgagatgata aggtttataa ggaatttgtt
      901 tgttctaatt tttcactcat tttgttctaa tttcttttaa caaatgttct ttttttttta
      961 gaacagttat gatatagtta gaatagttta aaataaggag tgagaaaaag atgaaagaaa
     1021 gatatggaac agtctataaa ggctctcaga ggctcataga cgaagaaagt ggagaagtca
     1081 tagaggtaga caagttatac cgtaaacaaa cgtctggtaa cttcgtaaag gcatatatag
     1141 tgcaattaat aagtatgtta gatatgattg gcggaaaaaa acttaaaatc gttaactata
     1201 tcctagataa tgtccactta agtaacaata caatgatagc tacaacaaga gaaatagcaa
     1261 aagctacagg aacaagtcta caaacagtaa taacaacact taaaatctta gaagaaggaa
     1321 atattataaa aagaaaaact ggagtattaa tgttaaaccc tgaactacta atgagaggcg
     1381 acgaccaaaa acaaaaatac ctcttactcg aatttgggaa ctttgagcaa gaggcaaatg
     1441 aaatagattg acctcccaat aacaccacgt agttattggg aggtcaatct atgaaatgcg
     1501 attaagggcc ggccagtggg caagttgaaa aattcacaaa aatgtggtat aatatctttg
     1561 ttcattagag cgataaactt gaatttgaga gggaacttag atggtatttg aaaaaattga
     1621 taaaaatagt tggaacagaa aagagtattt tgaccactac tttgcaagtg taccttgtac
     1681 ctacagcatg accgttaaag tggatatcac acaaataaag gaaaagggaa tgaaactata
     1741 tcctgcaatg ctttattata ttgcaatgat tgtaaaccgc cattcagagt ttaggacggc
     1801 aatcaatcaa gatggtgaat tggggatata tgatgagatg ataccaagct atacaatatt
     1861 tcacaatgat actgaaacat tttccagcct ttggactgag tgtaagtctg actttaaatc
     1921 atttttagca gattatgaaa gtgatacgca acggtatgga aacaatcata gaatggaagg
     1981 aaagccaaat gctccggaaa acatttttaa tgtatctatg ataccgtggt caaccttcga
     2041 tggctttaat ctgaatttgc agaaaggata tgattatttg attcctattt ttactatggg
     2101 gaaatattat aaagaagata acaaaattat acttcctttg gcaattcaag ttcatcacgc
     2161 agtatgtgac ggatttcaca tttgccgttt tgtaaacgaa ttgcaggaat tgataaatag
     2221 ttaacttcag gtttgtctgt aactaaaaac aagtatttaa gcaaaaacat cgtagaaata
     2281 cggtgttttt tgttacccta agtttaaact cctttttgat aatctcatga ccaaaatccc
     2341 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2401 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2461 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2521 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     2581 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2641 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2701 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2761 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2821 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2881 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2941 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3001 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3061 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3121 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     3181 acgcagggcc c
//
