LOCUS       V004437                 4353 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004437
VERSION     V004437
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4353)
  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 4353)
  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 4353)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4353)
  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..4353
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    9..306
                     /gene="pyrF fragment"
                     /note="mediates homologous recombination with the
                     chromosome of Clostridium acetobutylicum ATCC824"
     gene            9..306
                     /gene="pyrF fragment"
                     /label="pyrF fragment"
     primer_bind     complement(431..447)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             612..1349
                     /gene="pyrK"
                     /label="Dihydroorotate dehydrogenase B (NAD(+)), electron
                     transfer subunit"
                     /note="Dihydroorotate dehydrogenase B (NAD(+)), electron
                     transfer subunit from Clostridium acetobutylicum (strain
                     ATCC 824 / DSM 792 / JCM 1419 / LMG 5710 / VKM B-1787).
                     Accession#: Q97FS6"
     gene            1345..1794
                     /gene="pyrD fragment"
                     /label="pyrD fragment"
     CDS             2173..2613
                     /codon_start=1
                     /gene="repL"
                     /product="plasmid replication protein"
                     /label="repL"
                     /protein_id="AFA34925.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     gene            2173..2613
                     /gene="repL"
                     /label="repL"
     CDS             2763..3383
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      3563..4151
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 cctgcagggt gtttaggact tgatactgat attacttatg taccagaaga gttttgtaag
       61 aaatttaata gtatagaaga tgctatattt aattttaata agaaaataat tgatgcgact
      121 ttagatgttg tttcatgtta taaggtgcaa attgcatatt atgaagctta tggtttaaaa
      181 ggacttttgg cctataaaag gacacttgaa tatttgagag aaaaaaaggc tattgcaatt
      241 gctgatataa aaagaggaga tatagctaaa acagctgaaa tgtatgctaa agctcacttt
      301 gaaggttaat gagcggccgc tgtatccata tgaccatgat tacgaattcg agctcggtac
      361 ccggggatcc tctagagtcg acgtcacgcg tccatggaga tctcgaggcc tgcagacatg
      421 caagcttggc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
      481 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
      541 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc tagcattttg
      601 ggggaatttt gatgaaggaa aagtatacag tagaaaaagt atatgaaaat ataaaagttg
      661 aagatggtat atacaaactt agtataaagg gtgaatttga ggtgaggcca ggacaatttt
      721 atcttttaag agcttgggat atagaaccaa cactttctag acctattagt atatatgatg
      781 cagatgacga aaaaatatcg ttcttatact ctgttgttgg aaaaggaact gaaattttat
      841 ctaaattaaa gagcggcgat gaaataaaga taacaggacc tttaggaaat ggatttaacg
      901 taaaaaggat aagtggaaaa gtggctatag tttgtggtgg tataggtgta gcaccaatgg
      961 tatatctggc taaaaactta aaaaattgta atgttgattt ttatgctggc ttcaaaactg
     1021 tgagtaaaac tgtggataat gtggaaaaat atgttaaaga gttaaagttg tccacagaag
     1081 atggaagtat tggacataag gggtatgtaa cagataactt taagccagaa gaatacgatt
     1141 atgttttatg ctgcggacct gagataatga tgtataaagt tgttaaaatg tgtgaacaaa
     1201 agaatgttcc tgtatatatt tcaatggaga aaaaaatggc atgtggaata ggtgcatgcc
     1261 ttgtatgcac ttgtaaaact aagggtggaa gaagaagagc ttgtaaagag ggcccagtat
     1321 ttttaggaag tgagttgata ttaaatgact aaagtaaata tttgtggaat agattttaag
     1381 aaccccgtta ttgctgcttc tggcaccttt ggatttggag aagagtttag taagtatttt
     1441 gatgtttcaa ggcttggtgg catatcttca aagggactta cattgaatcc taaggaaggt
     1501 aatgatggtg caagagtttt tgaggtcaca ggcggaatga tgaatagtgt aggacttcaa
     1561 aatcctggag ttaaagagtt tataaaaaaa gaacttccta agatgaaaaa aatagataca
     1621 gtatgtattg ttaaccttgg tggaagctgt gaggatgatt atttaagggg catggagctt
     1681 cttgagaata cagatgctga tatgatagaa cttaatatat cctgtcctaa tgtaaagcac
     1741 ggcggcatgg cttttggaat aaaatcagaa gttgcttata atgttgtatc acaaggcgcg
     1801 ccgcattcac ttcttttcta tataaatatg agcgaagcga ataagcgtcg gaaaagcagc
     1861 aaaaagtttc ctttttgctg ttggagcatg ggggttcagg gggtgcagta tctgacgtca
     1921 atgccgagcg aaagcgagcc gaagggtagc atttacgtta gataaccccc tgatatgctc
     1981 cgacgcttta tatagaaaag aagattcaac taggtaaaat cttaatatag gttgagatga
     2041 taaggtttat aaggaatttg tttgttctaa tttttcactc attttgttct aatttctttt
     2101 aacaaatgtt cttttttttt tagaacagtt atgatatagt tagaatagtt taaaataagg
     2161 agtgagaaaa agatgaaaga aagatatgga acagtctata aaggctctca gaggctcata
     2221 gacgaagaaa gtggagaagt catagaggta gacaagttat accgtaaaca aacgtctggt
     2281 aacttcgtaa aggcatatat agtgcaatta ataagtatgt tagatatgat tggcggaaaa
     2341 aaacttaaaa tcgttaacta tatcctagat aatgtccact taagtaacaa tacaatgata
     2401 gctacaacaa gagaaatagc aaaagctaca ggaacaagtc tacaaacagt aataacaaca
     2461 cttaaaatct tagaagaagg aaatattata aaaagaaaaa ctggagtatt aatgttaaac
     2521 cctgaactac taatgagagg cgacgaccaa aaacaaaaat acctcttact cgaatttggg
     2581 aactttgagc aagaggcaaa tgaaatagat tgacctccca ataacaccac gtagttattg
     2641 ggaggtcaat ctatgaaatg cgattaaggg ccggccagtg ggcaagttga aaaattcaca
     2701 aaaatgtggt ataatatctt tgttcattag agcgataaac ttgaatttga gagggaactt
     2761 agatggtatt tgaaaaaatt gataaaaata gttggaacag aaaagagtat tttgaccact
     2821 actttgcaag tgtaccttgt acctacagca tgaccgttaa agtggatatc acacaaataa
     2881 aggaaaaggg aatgaaacta tatcctgcaa tgctttatta tattgcaatg attgtaaacc
     2941 gccattcaga gtttaggacg gcaatcaatc aagatggtga attggggata tatgatgaga
     3001 tgataccaag ctatacaata tttcacaatg atactgaaac attttccagc ctttggactg
     3061 agtgtaagtc tgactttaaa tcatttttag cagattatga aagtgatacg caacggtatg
     3121 gaaacaatca tagaatggaa ggaaagccaa atgctccgga aaacattttt aatgtatcta
     3181 tgataccgtg gtcaaccttc gatggcttta atctgaattt gcagaaagga tatgattatt
     3241 tgattcctat ttttactatg gggaaatatt ataaagaaga taacaaaatt atacttcctt
     3301 tggcaattca agttcatcac gcagtatgtg acggatttca catttgccgt tttgtaaacg
     3361 aattgcagga attgataaat agttaacttc aggtttgtct gtaactaaaa acaagtattt
     3421 aagcaaaaac atcgtagaaa tacggtgttt tttgttaccc taagtttaaa ctcctttttg
     3481 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3541 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3601 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3661 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     3721 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3781 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3841 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3901 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3961 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4021 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4081 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4141 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4201 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     4261 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     4321 aggaagcgga agagcgccca atacgcaggg ccc
//
