LOCUS       V004430                 3870 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004430
VERSION     V004430
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 3870)
  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 3870)
  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 3870)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3870)
  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..3870
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      9..62
                     /label="derived from CD0164 gene of Clostridium difficile"
                     /note="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"
     misc_feature    90..389
                     /gene="thl fragment"
                     /note="mediates homologous recombination with the
                     chromosome of Clostridium acetobutylicum ATCC824"
     gene            90..389
                     /gene="thl fragment"
                     /label="thl fragment"
     CDS             412..987
                     /codon_start=1
                     /gene="pyrE"
                     /product="orotate phosphoribosyltransferase"
                     /label="pyrE"
                     /protein_id="AFA34992.1"
                     /translation="MSNINVIDILKESNALLEGHFLLSSGRHSNRYCQCAKLLQYPQKA
                     EKVISVVAEKLKDADFNIIVGPAMGGVIVSYELARQTNRPGIFAERKDGIMCIRRGFEI
                     KKGDKVIISEDVITTGKSSLEVAKVIEEMGGEVVGIACIVDRRAEGIKTNYPIYSACKL
                     EIETYEKDNCELCKKNIPFVKPGSREQK"
     gene            412..987
                     /gene="pyrE"
                     /label="pyrE"
     primer_bind     complement(1106..1122)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     regulatory      1270..1311
                     /label="derived from fdx gene of Clostridium pasteurianum"
                     /note="derived from fdx gene of Clostridium pasteurianum"
                     /regulatory_class="terminator"
     CDS             1690..2130
                     /codon_start=1
                     /gene="repL"
                     /product="plasmid replication protein"
                     /label="repL"
                     /protein_id="AFA34994.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     gene            1690..2130
                     /gene="repL"
                     /label="repL"
     CDS             2280..2900
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      3080..3668
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 cctgcaggat aaaaaaattg tagataaatt ttataaaata gttttatcta caattttttt
       61 atcaggaaac agctatgacc gcggccatct atgcaacaaa agcagctatt gaaaaagcag
      121 gttggacagt tgatgaatta gatttaatag aatcaaatga agcttttgca gctcaaagtt
      181 tagcagtagc aaaagattta aaatttgata tgaataaagt aaatgtaaat ggaggagcta
      241 ttgcccttgg tcatccaatt ggagcatcag gtgcaagaat actcgttact cttgtacacg
      301 caatgcaaaa aagagatgca aaaaaaggct tagcaacttt atgtataggt ggcggacaag
      361 gaacagcaat attgctagaa aagtgctaga tcgattaagg agaagatata aatgagtaat
      421 ataaatgtta tagatatatt aaaagaatca aatgcattat tagaagggca tttcttacta
      481 tcatctggaa gacatagtaa tagatattgt caatgtgcga aactacttca atatccacaa
      541 aaagcagaaa aagttattag tgtagtagca gaaaagctta aagatgcaga ctttaatata
      601 atagtaggac cagctatggg tggtgtaata gtatcttatg aattagcaag gcaaactaat
      661 agaccaggca tatttgcaga aagaaaagat ggaattatgt gtatacgaag aggatttgaa
      721 ataaaaaaag gagataaagt tataatatca gaggatgtaa taactacagg aaaatcttct
      781 ttagaagtag ctaaagttat agaagaaatg ggtggagaag tagtaggtat agcatgcata
      841 gttgatagaa gagcggaagg tataaaaaca aactacccaa tatatagtgc ttgtaaatta
      901 gaaatagaaa cttatgaaaa agataattgt gagttatgta aaaaaaatat accttttgta
      961 aaaccaggta gtagagaaca aaaataagcg gccgctgtat ccatatgacc atgattacga
     1021 attcgagctc ggtacccggg gatcctctag agtcgacgtc acgcgtccat ggagatctcg
     1081 aggcctgcag acatgcaagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     1141 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     1201 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     1261 ggcgctagca taaaaataag aagcctgcat ttgcaggctt cttattttta tggcgcgccg
     1321 cattcacttc ttttctatat aaatatgagc gaagcgaata agcgtcggaa aagcagcaaa
     1381 aagtttcctt tttgctgttg gagcatgggg gttcaggggg tgcagtatct gacgtcaatg
     1441 ccgagcgaaa gcgagccgaa gggtagcatt tacgttagat aaccccctga tatgctccga
     1501 cgctttatat agaaaagaag attcaactag gtaaaatctt aatataggtt gagatgataa
     1561 ggtttataag gaatttgttt gttctaattt ttcactcatt ttgttctaat ttcttttaac
     1621 aaatgttctt ttttttttag aacagttatg atatagttag aatagtttaa aataaggagt
     1681 gagaaaaaga tgaaagaaag atatggaaca gtctataaag gctctcagag gctcatagac
     1741 gaagaaagtg gagaagtcat agaggtagac aagttatacc gtaaacaaac gtctggtaac
     1801 ttcgtaaagg catatatagt gcaattaata agtatgttag atatgattgg cggaaaaaaa
     1861 cttaaaatcg ttaactatat cctagataat gtccacttaa gtaacaatac aatgatagct
     1921 acaacaagag aaatagcaaa agctacagga acaagtctac aaacagtaat aacaacactt
     1981 aaaatcttag aagaaggaaa tattataaaa agaaaaactg gagtattaat gttaaaccct
     2041 gaactactaa tgagaggcga cgaccaaaaa caaaaatacc tcttactcga atttgggaac
     2101 tttgagcaag aggcaaatga aatagattga cctcccaata acaccacgta gttattggga
     2161 ggtcaatcta tgaaatgcga ttaagggccg gccagtgggc aagttgaaaa attcacaaaa
     2221 atgtggtata atatctttgt tcattagagc gataaacttg aatttgagag ggaacttaga
     2281 tggtatttga aaaaattgat aaaaatagtt ggaacagaaa agagtatttt gaccactact
     2341 ttgcaagtgt accttgtacc tacagcatga ccgttaaagt ggatatcaca caaataaagg
     2401 aaaagggaat gaaactatat cctgcaatgc tttattatat tgcaatgatt gtaaaccgcc
     2461 attcagagtt taggacggca atcaatcaag atggtgaatt ggggatatat gatgagatga
     2521 taccaagcta tacaatattt cacaatgata ctgaaacatt ttccagcctt tggactgagt
     2581 gtaagtctga ctttaaatca tttttagcag attatgaaag tgatacgcaa cggtatggaa
     2641 acaatcatag aatggaagga aagccaaatg ctccggaaaa catttttaat gtatctatga
     2701 taccgtggtc aaccttcgat ggctttaatc tgaatttgca gaaaggatat gattatttga
     2761 ttcctatttt tactatgggg aaatattata aagaagataa caaaattata cttcctttgg
     2821 caattcaagt tcatcacgca gtatgtgacg gatttcacat ttgccgtttt gtaaacgaat
     2881 tgcaggaatt gataaatagt taacttcagg tttgtctgta actaaaaaca agtatttaag
     2941 caaaaacatc gtagaaatac ggtgtttttt gttaccctaa gtttaaactc ctttttgata
     3001 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3061 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3121 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3181 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     3241 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3301 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3361 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3421 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3481 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3541 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3601 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3661 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3721 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     3781 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     3841 aagcggaaga gcgcccaata cgcagggccc
//
