LOCUS       V004439                 5862 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004439
VERSION     V004439
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5862)
  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 5862)
  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 5862)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5862)
  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..5862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    9..308
                     /gene="pyrE fragment"
                     /note="mediates homologous recombination with the
                     chromosome of Clostridium difficile 630"
     gene            9..308
                     /gene="pyrE fragment"
                     /label="pyrE fragment"
     primer_bind     complement(427..443)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    591..1790
                     /note="mediates homologous recombination with the
                     chromosome of Clostridium difficile 630"
     CDS             802..1728
                     /codon_start=1
                     /gene="CD0188"
                     /product="putative ketopantoate reductase"
                     /label="CD0188"
                     /protein_id="AFA34961.1"
                     /translation="MKNQLNENNKKVVFFGVGAVGATFAEQFFNSKYDFKILCDNERKK
                     RYLEEGFIINGKRYDFDYVTKDEYKQEADFIIIGLKYNNLKENIKELDGLVGKNTVIMS
                     LLNGVDSEEIIGERFGIEKMVYSYVTNIDAKKINNNIIHTTNGIIVFGNKDNSEDRKTN
                     IITEVFDDVNIEYTLSKDIQRDMWWKYMVNIGVNQTSAILGAPYGVFQSSEHLRELAKS
                     AMREVVAIAQAKDISLTEDDVEHSLHRILEHSKEGRTSMLQDVEAHRLTEVDMFSKNIC
                     KLGKKYNIPTPINQTFFYMIKVIESRF"
     gene            802..1728
                     /gene="CD0188"
                     /label="CD0188"
     CDS             2517..2834
                     /codon_start=1
                     /gene="repH"
                     /product="plasmid replication protein"
                     /label="repH"
                     /protein_id="AFA34962.1"
                     /translation="MKRRAYMKTCKNCKEFIKDTEICKIHSLMIHDKTVATYCSKYNES
                     RCLHKGKVQCINCSKMNRYGWCAIKMRCFTEEEQKKERTCIKYYARSFKKAHVKKSKKK
                     K"
     gene            2517..2834
                     /gene="repH"
                     /label="repH"
     CDS             3506..4126
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      4306..4894
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     oriT            5215..5324
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             5357..5725
                     /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 gacgttgtgc ggccgctgta tccatatgac catgattacg aattcgagct cggtacccgg
      361 ggatcctcta gagtcgacgt cacgcgtcca tggagatctc gaggcctgca gacatgcaag
      421 cttggcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
      481 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
      541 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctagc aaacttaatt
      601 atttatagtg ttacttaaaa aatgtaaatt ttttagagta aaaatttgta atataacata
      661 taacttaaaa ataaaaagca gttagcctta taaatgaagg ttagctgttt tttttgttaa
      721 aaatatattt acaaaataag agatagtagc agtacaatat aagtataagg taaaagtatt
      781 atatattagg gaggggtaag tatgaaaaat caattaaatg aaaataataa aaaggttgta
      841 ttttttggag taggtgctgt aggagctact tttgcagaac aattttttaa ctctaaatat
      901 gattttaaaa ttctttgtga taatgaaaga aagaaaagat atttagaaga aggatttata
      961 ataaatggaa aaaggtatga ttttgattat gtaactaaag atgagtataa acaagaggct
     1021 gattttataa ttataggtct aaaatataat aatttaaaag aaaatataaa agaattagat
     1081 ggattagttg gaaaaaatac agttataatg tctctgctaa atggagttga tagcgaagag
     1141 ataataggag aaagatttgg aattgaaaaa atggtatatt catatgttac caatatagat
     1201 gcaaagaaaa tcaataataa tattatacac actactaatg ggataattgt atttggtaac
     1261 aaagataata gtgaagatag aaaaactaat ataataaccg aagtctttga tgatgtaaat
     1321 atagaatata ctttatcaaa agatattcaa cgagatatgt ggtggaagta catggttaat
     1381 attggtgtaa atcaaacttc agctatactt ggtgcacctt atggagtttt tcagagttct
     1441 gagcatttaa gagaattagc aaaatctgca atgagggaag ttgttgctat agcacaagca
     1501 aaagacatat ctcttacaga agatgatgtg gaacattcat tacatagaat actagaacat
     1561 tcaaaagaag gaagaacatc aatgcttcaa gatgtggaag ctcatagact tacagaagta
     1621 gatatgtttt ctaagaatat ctgtaaactt ggaaaaaaat ataacatacc tactcctata
     1681 aatcagactt tcttttatat gataaaagta attgaaagta gattttaaag gggctttgta
     1741 agaaatattg ataattattg attttaaatt ttattaatgt tatatactat ggcgcgccgc
     1801 cattattttt ttgaacaatt gacaattcat ttcttatttt ttattaagtg atagtcaaaa
     1861 ggcataacag tgctgaatag aaagaaattt acagaaaaga aaattataga atttagtatg
     1921 attaattata ctcatttatg aatgtttaat tgaatacaaa aaaaaatact tgttatgtat
     1981 tcaattacgg gttaaaatat agacaagttg aaaaatttaa taaaaaaata agtcctcagc
     2041 tcttatatat taagctacca acttagtata taagccaaaa cttaaatgtg ctaccaacac
     2101 atcaagccgt tagagaactc tatctatagc aatatttcaa atgtaccgac atacaagaga
     2161 aacattaact atatatattc aatttatgag attatcttaa cagatataaa tgtaaattgc
     2221 aataagtaag atttagaagt ttatagcctt tgtgtattgg aagcagtacg caaaggcttt
     2281 tttatttgat aaaaattaga agtatattta ttttttcata attaatttat gaaaatgaaa
     2341 gggggtgagc aaagtgacag aggaaagcag tatcttatca aataacaagg tattagcaat
     2401 atcattattg actttagcag taaacattat gacttttata gtgcttgtag ctaagtagta
     2461 cgaaaggggg agctttaaaa agctccttgg aatacataga attcataaat taatttatga
     2521 aaagaagggc gtatatgaaa acttgtaaaa attgcaaaga gtttattaaa gatactgaaa
     2581 tatgcaaaat acattcgttg atgattcatg ataaaacagt agcaacctat tgcagtaaat
     2641 acaatgagtc aagatgttta cataaaggga aagtccaatg tattaattgt tcaaagatga
     2701 accgatatgg atggtgtgcc ataaaaatga gatgttttac agaggaagaa cagaaaaaag
     2761 aacgtacatg cattaaatat tatgcaagga gctttaaaaa agctcatgta aagaagagta
     2821 aaaagaaaaa ataatttatt tattaattta atattgagag tgccgacaca gtatgcacta
     2881 aaaaatatat ctgtggtgta gtgagccgat acaaaaggat agtcactcgc attttcataa
     2941 tacatcttat gttatgatta tgtgtcggtg ggacttcacg acgaaaaccc acaataaaaa
     3001 aagagttcgg ggtagggtta agcatagttg aggcaactaa acaatcaagc taggatatgc
     3061 agtagcagac cgtaaggtcg ttgtttaggt gtgttgtaat acatacgcta ttaagatgta
     3121 aaaatacgga taccaatgaa gggaaaagta taatttttgg atgtagtttg tttgttcatc
     3181 tatgggcaaa ctacgtccaa agccgtttcc aaatctgcta aaaagtatat cctttctaaa
     3241 atcaaagtca agtatgaaat cataaataaa gtttaatttt gaagttatta tgatattatg
     3301 tttttctatt aaaataaatt aagtatatag aatagtttaa taatagtata tacttaatgt
     3361 gataagtgtc tgacagtgtc acagaaagga tgattgttat ggattataag cggccggcca
     3421 gtgggcaagt tgaaaaattc acaaaaatgt ggtataatat ctttgttcat tagagcgata
     3481 aacttgaatt tgagagggaa cttagatggt atttgaaaaa attgataaaa atagttggaa
     3541 cagaaaagag tattttgacc actactttgc aagtgtacct tgtacctaca gcatgaccgt
     3601 taaagtggat atcacacaaa taaaggaaaa gggaatgaaa ctatatcctg caatgcttta
     3661 ttatattgca atgattgtaa accgccattc agagtttagg acggcaatca atcaagatgg
     3721 tgaattgggg atatatgatg agatgatacc aagctataca atatttcaca atgatactga
     3781 aacattttcc agcctttgga ctgagtgtaa gtctgacttt aaatcatttt tagcagatta
     3841 tgaaagtgat acgcaacggt atggaaacaa tcatagaatg gaaggaaagc caaatgctcc
     3901 ggaaaacatt tttaatgtat ctatgatacc gtggtcaacc ttcgatggct ttaatctgaa
     3961 tttgcagaaa ggatatgatt atttgattcc tatttttact atggggaaat attataaaga
     4021 agataacaaa attatacttc ctttggcaat tcaagttcat cacgcagtat gtgacggatt
     4081 tcacatttgc cgttttgtaa acgaattgca ggaattgata aatagttaac ttcaggtttg
     4141 tctgtaacta aaaacaagta tttaagcaaa aacatcgtag aaatacggtg ttttttgtta
     4201 ccctaagttt aaactccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4261 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4321 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4381 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4441 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4501 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4561 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4621 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4681 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4741 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4801 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4861 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4921 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4981 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     5041 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca gggccccctg
     5101 cttcggggtc attatagcga ttttttcggt atatccatcc tttttcgcac gatatacagg
     5161 attttgccaa agggttcgtg tagactttcc ttggtgtatc caacggcgtc agccgggcag
     5221 gataggtgaa gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca
     5281 cctggcggtg ctcaacggga atcctgctct gcgaggctgg ccggctaccg ccggcgtaac
     5341 agatgagggc aagcggatgg ctgatgaaac caagccaacc aggaagggca gcccacctat
     5401 caaggtgtac tgccttccag acgaacgaag agcgattgag gaaaaggcgg cggcggccgg
     5461 catgagcctg tcggcctacc tgctggccgt cggccagggc tacaaaatca cgggcgtcgt
     5521 ggactatgag cacgtccgcg agctggcccg catcaatggc gacctgggcc gcctgggcgg
     5581 cctgctgaaa ctctggctca ccgacgaccc gcgcacggcg cggttcggtg atgccacgat
     5641 cctcgccctg ctggcgaaga tcgaagagaa gcaggacgag cttggcaagg tcatgatggg
     5701 cgtggtccgc ccgagggcag agccatgact tttttagccg ctaaaacggc cggggggtgc
     5761 gcgtgattgc caagcacgtc cccatgcgct ccatcaagaa gagcgacttc gcggagctgg
     5821 tgaagtacat caccgacgag caaggcaaga ccgatcgggc cc
//
