LOCUS       V004428                 4863 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004428
VERSION     V004428
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4863)
  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 4863)
  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 4863)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4863)
  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..4863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            9..822
                     /gene="pyrF fragment"
                     /label="pyrF fragment"
     misc_feature    9..306
                     /gene="pyrF fragment"
                     /note="mediates homologous recombination with the
                     chromosome of Clostridium acetobutylicum ATCC824"
     primer_bind     complement(941..957)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             1122..1859
                     /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            1855..2304
                     /gene="pyrD fragment"
                     /label="pyrD fragment"
     CDS             2683..3123
                     /codon_start=1
                     /gene="repL"
                     /product="plasmid replication protein"
                     /label="repL"
                     /protein_id="AFA34931.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     gene            2683..3123
                     /gene="repL"
                     /label="repL"
     CDS             3273..3893
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      4073..4661
                     /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 gaaggtgatt ttgaagcgga ttttgttacg ttaaatcctt acatggggtt agatggtata
      361 gagccttata tgccttatat tgaaaaaatg gaaaaaggat tatttatttt gcttagaaca
      421 tcgaataaag gagcctatga tatacaatat ataaagactc agggcggaaa aaacgtatat
      481 gatgaggttg gagaaaaaat atatgattta ggtcaaaagg ctacgggaag gagcaagtat
      541 tcttcaatag gagcagtagt tggatgtact cacgttgaag aaggcgttga aattagaaat
      601 aaatttaaaa atatgttttt tctaattcca ggctatggag cacaaggtgg aactgcaaag
      661 gaagtaagtt tgtatttaag agaaggtaat ggtggagtgg taaattcctc aaggggaata
      721 cttcttgctt ataaaaaaga agaaaacggt gaaaaaatat ttgatgagtg tgcaaggctt
      781 gcagcgatta atatgagaga cgagatcaga aaaactttat gagcggccgc tgtatccata
      841 tgaccatgat tacgaattcg agctcggtac ccggggatcc tctagagtcg acgtcacgcg
      901 tccatggaga tctcgaggcc tgcagacatg caagcttggc actggccgtc gttttacaac
      961 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     1021 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     1081 gcctgaatgg cgaatggcgc tagcattttg ggggaatttt gatgaaggaa aagtatacag
     1141 tagaaaaagt atatgaaaat ataaaagttg aagatggtat atacaaactt agtataaagg
     1201 gtgaatttga ggtgaggcca ggacaatttt atcttttaag agcttgggat atagaaccaa
     1261 cactttctag acctattagt atatatgatg cagatgacga aaaaatatcg ttcttatact
     1321 ctgttgttgg aaaaggaact gaaattttat ctaaattaaa gagcggcgat gaaataaaga
     1381 taacaggacc tttaggaaat ggatttaacg taaaaaggat aagtggaaaa gtggctatag
     1441 tttgtggtgg tataggtgta gcaccaatgg tatatctggc taaaaactta aaaaattgta
     1501 atgttgattt ttatgctggc ttcaaaactg tgagtaaaac tgtggataat gtggaaaaat
     1561 atgttaaaga gttaaagttg tccacagaag atggaagtat tggacataag gggtatgtaa
     1621 cagataactt taagccagaa gaatacgatt atgttttatg ctgcggacct gagataatga
     1681 tgtataaagt tgttaaaatg tgtgaacaaa agaatgttcc tgtatatatt tcaatggaga
     1741 aaaaaatggc atgtggaata ggtgcatgcc ttgtatgcac ttgtaaaact aagggtggaa
     1801 gaagaagagc ttgtaaagag ggcccagtat ttttaggaag tgagttgata ttaaatgact
     1861 aaagtaaata tttgtggaat agattttaag aaccccgtta ttgctgcttc tggcaccttt
     1921 ggatttggag aagagtttag taagtatttt gatgtttcaa ggcttggtgg catatcttca
     1981 aagggactta cattgaatcc taaggaaggt aatgatggtg caagagtttt tgaggtcaca
     2041 ggcggaatga tgaatagtgt aggacttcaa aatcctggag ttaaagagtt tataaaaaaa
     2101 gaacttccta agatgaaaaa aatagataca gtatgtattg ttaaccttgg tggaagctgt
     2161 gaggatgatt atttaagggg catggagctt cttgagaata cagatgctga tatgatagaa
     2221 cttaatatat cctgtcctaa tgtaaagcac ggcggcatgg cttttggaat aaaatcagaa
     2281 gttgcttata atgttgtatc acaaggcgcg ccgcattcac ttcttttcta tataaatatg
     2341 agcgaagcga ataagcgtcg gaaaagcagc aaaaagtttc ctttttgctg ttggagcatg
     2401 ggggttcagg gggtgcagta tctgacgtca atgccgagcg aaagcgagcc gaagggtagc
     2461 atttacgtta gataaccccc tgatatgctc cgacgcttta tatagaaaag aagattcaac
     2521 taggtaaaat cttaatatag gttgagatga taaggtttat aaggaatttg tttgttctaa
     2581 tttttcactc attttgttct aatttctttt aacaaatgtt cttttttttt tagaacagtt
     2641 atgatatagt tagaatagtt taaaataagg agtgagaaaa agatgaaaga aagatatgga
     2701 acagtctata aaggctctca gaggctcata gacgaagaaa gtggagaagt catagaggta
     2761 gacaagttat accgtaaaca aacgtctggt aacttcgtaa aggcatatat agtgcaatta
     2821 ataagtatgt tagatatgat tggcggaaaa aaacttaaaa tcgttaacta tatcctagat
     2881 aatgtccact taagtaacaa tacaatgata gctacaacaa gagaaatagc aaaagctaca
     2941 ggaacaagtc tacaaacagt aataacaaca cttaaaatct tagaagaagg aaatattata
     3001 aaaagaaaaa ctggagtatt aatgttaaac cctgaactac taatgagagg cgacgaccaa
     3061 aaacaaaaat acctcttact cgaatttggg aactttgagc aagaggcaaa tgaaatagat
     3121 tgacctccca ataacaccac gtagttattg ggaggtcaat ctatgaaatg cgattaaggg
     3181 ccggccagtg ggcaagttga aaaattcaca aaaatgtggt ataatatctt tgttcattag
     3241 agcgataaac ttgaatttga gagggaactt agatggtatt tgaaaaaatt gataaaaata
     3301 gttggaacag aaaagagtat tttgaccact actttgcaag tgtaccttgt acctacagca
     3361 tgaccgttaa agtggatatc acacaaataa aggaaaaggg aatgaaacta tatcctgcaa
     3421 tgctttatta tattgcaatg attgtaaacc gccattcaga gtttaggacg gcaatcaatc
     3481 aagatggtga attggggata tatgatgaga tgataccaag ctatacaata tttcacaatg
     3541 atactgaaac attttccagc ctttggactg agtgtaagtc tgactttaaa tcatttttag
     3601 cagattatga aagtgatacg caacggtatg gaaacaatca tagaatggaa ggaaagccaa
     3661 atgctccgga aaacattttt aatgtatcta tgataccgtg gtcaaccttc gatggcttta
     3721 atctgaattt gcagaaagga tatgattatt tgattcctat ttttactatg gggaaatatt
     3781 ataaagaaga taacaaaatt atacttcctt tggcaattca agttcatcac gcagtatgtg
     3841 acggatttca catttgccgt tttgtaaacg aattgcagga attgataaat agttaacttc
     3901 aggtttgtct gtaactaaaa acaagtattt aagcaaaaac atcgtagaaa tacggtgttt
     3961 tttgttaccc taagtttaaa ctcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4021 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4081 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4141 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4201 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4261 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4321 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4381 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4441 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4501 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4561 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4621 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4681 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4741 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4801 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaggg
     4861 ccc
//
