LOCUS       V004447                 3195 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004447
VERSION     V004447
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 3195)
  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 3195)
  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 3195)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3195)
  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..3195
                     /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"
     regulatory      595..636
                     /label="derived from fdx gene of Clostridium pasteurianum"
                     /note="derived from fdx gene of Clostridium pasteurianum"
                     /regulatory_class="terminator"
     CDS             1015..1455
                     /codon_start=1
                     /gene="repL"
                     /product="plasmid replication protein"
                     /label="repL"
                     /protein_id="AFA34922.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     gene            1015..1455
                     /gene="repL"
                     /label="repL"
     CDS             1605..2225
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      2405..2993
                     /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 tagcataaaa
      601 ataagaagcc tgcatttgca ggcttcttat ttttatggcg cgccgcattc acttcttttc
      661 tatataaata tgagcgaagc gaataagcgt cggaaaagca gcaaaaagtt tcctttttgc
      721 tgttggagca tgggggttca gggggtgcag tatctgacgt caatgccgag cgaaagcgag
      781 ccgaagggta gcatttacgt tagataaccc cctgatatgc tccgacgctt tatatagaaa
      841 agaagattca actaggtaaa atcttaatat aggttgagat gataaggttt ataaggaatt
      901 tgtttgttct aatttttcac tcattttgtt ctaatttctt ttaacaaatg ttcttttttt
      961 tttagaacag ttatgatata gttagaatag tttaaaataa ggagtgagaa aaagatgaaa
     1021 gaaagatatg gaacagtcta taaaggctct cagaggctca tagacgaaga aagtggagaa
     1081 gtcatagagg tagacaagtt ataccgtaaa caaacgtctg gtaacttcgt aaaggcatat
     1141 atagtgcaat taataagtat gttagatatg attggcggaa aaaaacttaa aatcgttaac
     1201 tatatcctag ataatgtcca cttaagtaac aatacaatga tagctacaac aagagaaata
     1261 gcaaaagcta caggaacaag tctacaaaca gtaataacaa cacttaaaat cttagaagaa
     1321 ggaaatatta taaaaagaaa aactggagta ttaatgttaa accctgaact actaatgaga
     1381 ggcgacgacc aaaaacaaaa atacctctta ctcgaatttg ggaactttga gcaagaggca
     1441 aatgaaatag attgacctcc caataacacc acgtagttat tgggaggtca atctatgaaa
     1501 tgcgattaag ggccggccag tgggcaagtt gaaaaattca caaaaatgtg gtataatatc
     1561 tttgttcatt agagcgataa acttgaattt gagagggaac ttagatggta tttgaaaaaa
     1621 ttgataaaaa tagttggaac agaaaagagt attttgacca ctactttgca agtgtacctt
     1681 gtacctacag catgaccgtt aaagtggata tcacacaaat aaaggaaaag ggaatgaaac
     1741 tatatcctgc aatgctttat tatattgcaa tgattgtaaa ccgccattca gagtttagga
     1801 cggcaatcaa tcaagatggt gaattgggga tatatgatga gatgatacca agctatacaa
     1861 tatttcacaa tgatactgaa acattttcca gcctttggac tgagtgtaag tctgacttta
     1921 aatcattttt agcagattat gaaagtgata cgcaacggta tggaaacaat catagaatgg
     1981 aaggaaagcc aaatgctccg gaaaacattt ttaatgtatc tatgataccg tggtcaacct
     2041 tcgatggctt taatctgaat ttgcagaaag gatatgatta tttgattcct atttttacta
     2101 tggggaaata ttataaagaa gataacaaaa ttatacttcc tttggcaatt caagttcatc
     2161 acgcagtatg tgacggattt cacatttgcc gttttgtaaa cgaattgcag gaattgataa
     2221 atagttaact tcaggtttgt ctgtaactaa aaacaagtat ttaagcaaaa acatcgtaga
     2281 aatacggtgt tttttgttac cctaagttta aactcctttt tgataatctc atgaccaaaa
     2341 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2401 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2461 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2521 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     2581 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2641 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2701 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2761 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2821 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2881 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2941 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3001 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     3061 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     3121 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     3181 caatacgcag ggccc
//
