LOCUS       V004443                 4687 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004443
VERSION     V004443
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4687)
  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 4687)
  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 4687)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4687)
  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..4687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            9..646
                     /gene="pyrE fragment"
                     /label="pyrE fragment"
     misc_feature    9..308
                     /gene="pyrE fragment"
                     /note="mediates homologous recombination with the
                     chromosome of Clostridium acetobutylicum ATCC824"
     primer_bind     complement(765..781)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    929..2128
                     /note="mediates homologous recombination with the
                     chromosome of Clostridium acetobutylicum ATCC824"
     gene            complement(979..2128)
                     /gene="hydA fragment"
                     /label="hydA fragment"
     CDS             2507..2947
                     /codon_start=1
                     /gene="repL"
                     /product="plasmid replication protein"
                     /label="repL"
                     /protein_id="AFA34945.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     gene            2507..2947
                     /gene="repL"
                     /label="repL"
     CDS             3097..3717
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      3897..4485
                     /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 acttcttgga gcaaagcttg aagaaggaga cagagttata attgtagaag atgtcacaac
      361 agctggtaca tcagtatacg aaacaatgcc tatacttaaa tcacaggctg aggttgatgt
      421 aaagggaatc ataatatcag tggatagaat ggaaagaggt aagggagata agagtgcctt
      481 aactgaactt aaagaaaagt ttggatttaa aacatgttct attgttacta tggaagaggt
      541 agtagaatat ttgtataaga aaaatatcaa tggcaaagta atcatagatg ataaaatgaa
      601 agatagaatt aatgagtact ataaagagta tggagtaaaa tagtaagcgg ccgctgtatc
      661 catatgacca tgattacgaa ttcgagctcg gtacccgggg atcctctaga gtcgacgtca
      721 cgcgtccatg gagatctcga ggcctgcaga catgcaagct tggcactggc cgtcgtttta
      781 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
      841 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
      901 cgcagcctga atggcgaatg gcgctagcta aaataaatgt gcctcaactt agatgttaag
      961 gcacatttat tttatatatt attcatgttt tgaaacattt ttatcttttg tgtattttac
     1021 gtgtagtaat ttgtgagcaa gtccttcacc tggttttcca aagtagctat catacatttt
     1081 aataatagct ggattatcat gtgactttct ctttgaaaga acatttttat cttggttgta
     1141 taatactgat gctcttagtt ttctgtaatc aacattttct ctatcaagag catttacgtg
     1201 aggttgacct ccaccattta tacatccacc agggcaagcc attacttcta taaagtgata
     1261 ttgtttttcg ttcatttttc cagatttcat aaactcgaag aagttagaag caccatttat
     1321 aacagcaacg tttagtttat ttccagcaat ttcaacttcc gcttctttta tgcctttaaa
     1381 gcctcttact tcagtgtaat caacattttc aagttcttta ttttcagcaa agtctttagc
     1441 tgatcttatt gcagcttcca taacgccacc ggttgcacca aagatagctc cagcaccact
     1501 gtaagtaccc atagcaggat caacttcacc atcttcaaga tctgcaaatt taatttttgc
     1561 atctttaatc atttttgcaa gctctcttgt agttaaggat gcatcaatat ctcttaagct
     1621 gttagtttcc atgaaaggaa tatctgcttc atatttttta tcattacaag gcatgatagt
     1681 aactgtataa acatcttctg gagctattcc tgaaattgaa ggatagtaag tttttgatgc
     1741 agtaccaaat atttgttgtg gtgattttgc tgatgaaaga ttatctaata attcaggatg
     1801 ataattttga gctaatctta cccatgcagg acagcaagat gtaaacatag ggaatgggcc
     1861 attattttta actctgccta aaagttcagt agcttcttcc attatagtca tatctgcacc
     1921 aaagtttata tcaaatactt tatcaaagcc taacattcta agtgcagtat atagttttcc
     1981 tgttacatct tttccatatc ccattttgaa taattcgccc atagcagttc ttactgatgg
     2041 agccattgca acaatgacat gttttttagg gtcattaaga gcttcttgaa ctttttctat
     2101 atgggatttt tcttttaaag cagcaacagg cgcgccgcat tcacttcttt tctatataaa
     2161 tatgagcgaa gcgaataagc gtcggaaaag cagcaaaaag tttccttttt gctgttggag
     2221 catgggggtt cagggggtgc agtatctgac gtcaatgccg agcgaaagcg agccgaaggg
     2281 tagcatttac gttagataac cccctgatat gctccgacgc tttatataga aaagaagatt
     2341 caactaggta aaatcttaat ataggttgag atgataaggt ttataaggaa tttgtttgtt
     2401 ctaatttttc actcattttg ttctaatttc ttttaacaaa tgttcttttt tttttagaac
     2461 agttatgata tagttagaat agtttaaaat aaggagtgag aaaaagatga aagaaagata
     2521 tggaacagtc tataaaggct ctcagaggct catagacgaa gaaagtggag aagtcataga
     2581 ggtagacaag ttataccgta aacaaacgtc tggtaacttc gtaaaggcat atatagtgca
     2641 attaataagt atgttagata tgattggcgg aaaaaaactt aaaatcgtta actatatcct
     2701 agataatgtc cacttaagta acaatacaat gatagctaca acaagagaaa tagcaaaagc
     2761 tacaggaaca agtctacaaa cagtaataac aacacttaaa atcttagaag aaggaaatat
     2821 tataaaaaga aaaactggag tattaatgtt aaaccctgaa ctactaatga gaggcgacga
     2881 ccaaaaacaa aaatacctct tactcgaatt tgggaacttt gagcaagagg caaatgaaat
     2941 agattgacct cccaataaca ccacgtagtt attgggaggt caatctatga aatgcgatta
     3001 agggccggcc agtgggcaag ttgaaaaatt cacaaaaatg tggtataata tctttgttca
     3061 ttagagcgat aaacttgaat ttgagaggga acttagatgg tatttgaaaa aattgataaa
     3121 aatagttgga acagaaaaga gtattttgac cactactttg caagtgtacc ttgtacctac
     3181 agcatgaccg ttaaagtgga tatcacacaa ataaaggaaa agggaatgaa actatatcct
     3241 gcaatgcttt attatattgc aatgattgta aaccgccatt cagagtttag gacggcaatc
     3301 aatcaagatg gtgaattggg gatatatgat gagatgatac caagctatac aatatttcac
     3361 aatgatactg aaacattttc cagcctttgg actgagtgta agtctgactt taaatcattt
     3421 ttagcagatt atgaaagtga tacgcaacgg tatggaaaca atcatagaat ggaaggaaag
     3481 ccaaatgctc cggaaaacat ttttaatgta tctatgatac cgtggtcaac cttcgatggc
     3541 tttaatctga atttgcagaa aggatatgat tatttgattc ctatttttac tatggggaaa
     3601 tattataaag aagataacaa aattatactt cctttggcaa ttcaagttca tcacgcagta
     3661 tgtgacggat ttcacatttg ccgttttgta aacgaattgc aggaattgat aaatagttaa
     3721 cttcaggttt gtctgtaact aaaaacaagt atttaagcaa aaacatcgta gaaatacggt
     3781 gttttttgtt accctaagtt taaactcctt tttgataatc tcatgaccaa aatcccttaa
     3841 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3901 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3961 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4021 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4081 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4141 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4201 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4261 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4321 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4381 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4441 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4501 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4561 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4621 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     4681 agggccc
//
