LOCUS       V004438                 4942 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004438
VERSION     V004438
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4942)
  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 4942)
  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 4942)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4942)
  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..4942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            9..546
                     /gene="pyrE fragment"
                     /label="pyrE fragment"
     misc_feature    9..308
                     /gene="pyrE fragment"
                     /note="mediates homologous recombination with the
                     chromosome of Clostridium difficile 630"
     primer_bind     complement(665..681)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     regulatory      829..870
                     /label="derived from fdx gene of Clostridium pasteurianum"
                     /note="derived from fdx gene of Clostridium pasteurianum"
                     /regulatory_class="terminator"
     CDS             1597..1914
                     /codon_start=1
                     /gene="repH"
                     /product="plasmid replication protein"
                     /label="repH"
                     /protein_id="AFA34966.1"
                     /translation="MKRRAYMKTCKNCKEFIKDTEICKIHSLMIHDKTVATYCSKYNES
                     RCLHKGKVQCINCSKMNRYGWCAIKMRCFTEEEQKKERTCIKYYARSFKKAHVKKSKKK
                     K"
     gene            1597..1914
                     /gene="repH"
                     /label="repH"
     CDS             2586..3206
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      3386..3974
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     oriT            4295..4404
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             4437..4805
                     /label="traJ"
                     /note="oriT-recognizing protein"
ORIGIN
        1 cctgcagggg agggacattt tttattatct tcaggaaaac acagtaatag atacgtacaa
       61 tgtgcaaaag tattaagatt cccagaatat gctgctgagg tattaagtac agttgttgaa
      121 caaataaaag acttagatat agacttagta gtaggaccag ctatgggtgg agtaatagtt
      181 tcttatgagt taggaagaca attaggaaaa gaagctgtat ttactgagag aaaagacaat
      241 acaatggagt taagaagagg atttgaagtt aaaaaaggag caaagataat aattgctgaa
      301 gacgttgtaa ctactggtaa atcaactatg gagacaaaaa gagtattaga agccttaggt
      361 ggagaagttg taggtgttgc atgtatagca gatagaacta atcatgatat aggtatgcct
      421 atatatagtg ctataaaact tgatattcaa gtttatgaat ctgatgagtg tcctttatgt
      481 aaggaaggaa aattaccagt tgttaagcct ggaagtagag agttcaagga attagggatg
      541 taataagcgg ccgctgtatc catatgacca tgattacgaa ttcgagctcg gtacccgggg
      601 atcctctaga gtcgacgtca cgcgtccatg gagatctcga ggcctgcaga catgcaagct
      661 tggcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
      721 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
      781 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgctagcat aaaaataaga
      841 agcctgcatt tgcaggcttc ttatttttat ggcgcgccgc cattattttt ttgaacaatt
      901 gacaattcat ttcttatttt ttattaagtg atagtcaaaa ggcataacag tgctgaatag
      961 aaagaaattt acagaaaaga aaattataga atttagtatg attaattata ctcatttatg
     1021 aatgtttaat tgaatacaaa aaaaaatact tgttatgtat tcaattacgg gttaaaatat
     1081 agacaagttg aaaaatttaa taaaaaaata agtcctcagc tcttatatat taagctacca
     1141 acttagtata taagccaaaa cttaaatgtg ctaccaacac atcaagccgt tagagaactc
     1201 tatctatagc aatatttcaa atgtaccgac atacaagaga aacattaact atatatattc
     1261 aatttatgag attatcttaa cagatataaa tgtaaattgc aataagtaag atttagaagt
     1321 ttatagcctt tgtgtattgg aagcagtacg caaaggcttt tttatttgat aaaaattaga
     1381 agtatattta ttttttcata attaatttat gaaaatgaaa gggggtgagc aaagtgacag
     1441 aggaaagcag tatcttatca aataacaagg tattagcaat atcattattg actttagcag
     1501 taaacattat gacttttata gtgcttgtag ctaagtagta cgaaaggggg agctttaaaa
     1561 agctccttgg aatacataga attcataaat taatttatga aaagaagggc gtatatgaaa
     1621 acttgtaaaa attgcaaaga gtttattaaa gatactgaaa tatgcaaaat acattcgttg
     1681 atgattcatg ataaaacagt agcaacctat tgcagtaaat acaatgagtc aagatgttta
     1741 cataaaggga aagtccaatg tattaattgt tcaaagatga accgatatgg atggtgtgcc
     1801 ataaaaatga gatgttttac agaggaagaa cagaaaaaag aacgtacatg cattaaatat
     1861 tatgcaagga gctttaaaaa agctcatgta aagaagagta aaaagaaaaa ataatttatt
     1921 tattaattta atattgagag tgccgacaca gtatgcacta aaaaatatat ctgtggtgta
     1981 gtgagccgat acaaaaggat agtcactcgc attttcataa tacatcttat gttatgatta
     2041 tgtgtcggtg ggacttcacg acgaaaaccc acaataaaaa aagagttcgg ggtagggtta
     2101 agcatagttg aggcaactaa acaatcaagc taggatatgc agtagcagac cgtaaggtcg
     2161 ttgtttaggt gtgttgtaat acatacgcta ttaagatgta aaaatacgga taccaatgaa
     2221 gggaaaagta taatttttgg atgtagtttg tttgttcatc tatgggcaaa ctacgtccaa
     2281 agccgtttcc aaatctgcta aaaagtatat cctttctaaa atcaaagtca agtatgaaat
     2341 cataaataaa gtttaatttt gaagttatta tgatattatg tttttctatt aaaataaatt
     2401 aagtatatag aatagtttaa taatagtata tacttaatgt gataagtgtc tgacagtgtc
     2461 acagaaagga tgattgttat ggattataag cggccggcca gtgggcaagt tgaaaaattc
     2521 acaaaaatgt ggtataatat ctttgttcat tagagcgata aacttgaatt tgagagggaa
     2581 cttagatggt atttgaaaaa attgataaaa atagttggaa cagaaaagag tattttgacc
     2641 actactttgc aagtgtacct tgtacctaca gcatgaccgt taaagtggat atcacacaaa
     2701 taaaggaaaa gggaatgaaa ctatatcctg caatgcttta ttatattgca atgattgtaa
     2761 accgccattc agagtttagg acggcaatca atcaagatgg tgaattgggg atatatgatg
     2821 agatgatacc aagctataca atatttcaca atgatactga aacattttcc agcctttgga
     2881 ctgagtgtaa gtctgacttt aaatcatttt tagcagatta tgaaagtgat acgcaacggt
     2941 atggaaacaa tcatagaatg gaaggaaagc caaatgctcc ggaaaacatt tttaatgtat
     3001 ctatgatacc gtggtcaacc ttcgatggct ttaatctgaa tttgcagaaa ggatatgatt
     3061 atttgattcc tatttttact atggggaaat attataaaga agataacaaa attatacttc
     3121 ctttggcaat tcaagttcat cacgcagtat gtgacggatt tcacatttgc cgttttgtaa
     3181 acgaattgca ggaattgata aatagttaac ttcaggtttg tctgtaacta aaaacaagta
     3241 tttaagcaaa aacatcgtag aaatacggtg ttttttgtta ccctaagttt aaactccttt
     3301 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3361 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3421 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3481 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3541 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3601 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3661 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3721 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3781 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3841 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3901 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3961 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4021 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4081 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4141 gcgaggaagc ggaagagcgc ccaatacgca gggccccctg cttcggggtc attatagcga
     4201 ttttttcggt atatccatcc tttttcgcac gatatacagg attttgccaa agggttcgtg
     4261 tagactttcc ttggtgtatc caacggcgtc agccgggcag gataggtgaa gtaggcccac
     4321 ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca cctggcggtg ctcaacggga
     4381 atcctgctct gcgaggctgg ccggctaccg ccggcgtaac agatgagggc aagcggatgg
     4441 ctgatgaaac caagccaacc aggaagggca gcccacctat caaggtgtac tgccttccag
     4501 acgaacgaag agcgattgag gaaaaggcgg cggcggccgg catgagcctg tcggcctacc
     4561 tgctggccgt cggccagggc tacaaaatca cgggcgtcgt ggactatgag cacgtccgcg
     4621 agctggcccg catcaatggc gacctgggcc gcctgggcgg cctgctgaaa ctctggctca
     4681 ccgacgaccc gcgcacggcg cggttcggtg atgccacgat cctcgccctg ctggcgaaga
     4741 tcgaagagaa gcaggacgag cttggcaagg tcatgatggg cgtggtccgc ccgagggcag
     4801 agccatgact tttttagccg ctaaaacggc cggggggtgc gcgtgattgc caagcacgtc
     4861 cccatgcgct ccatcaagaa gagcgacttc gcggagctgg tgaagtacat caccgacgag
     4921 caaggcaaga ccgatcgggc cc
//
