LOCUS       V004440                 4704 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004440
VERSION     V004440
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4704)
  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 4704)
  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 4704)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4704)
  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..4704
                     /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"
     regulatory      591..632
                     /label="derived from fdx gene of Clostridium pasteurianum"
                     /note="derived from fdx gene of Clostridium pasteurianum"
                     /regulatory_class="terminator"
     CDS             1359..1676
                     /codon_start=1
                     /gene="repH"
                     /product="plasmid replication protein"
                     /label="repH"
                     /protein_id="AFA34957.1"
                     /translation="MKRRAYMKTCKNCKEFIKDTEICKIHSLMIHDKTVATYCSKYNES
                     RCLHKGKVQCINCSKMNRYGWCAIKMRCFTEEEQKKERTCIKYYARSFKKAHVKKSKKK
                     K"
     gene            1359..1676
                     /gene="repH"
                     /label="repH"
     CDS             2348..2968
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      3148..3736
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     oriT            4057..4166
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             4199..4567
                     /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 ataaaaataa
      601 gaagcctgca tttgcaggct tcttattttt atggcgcgcc gccattattt ttttgaacaa
      661 ttgacaattc atttcttatt ttttattaag tgatagtcaa aaggcataac agtgctgaat
      721 agaaagaaat ttacagaaaa gaaaattata gaatttagta tgattaatta tactcattta
      781 tgaatgttta attgaataca aaaaaaaata cttgttatgt attcaattac gggttaaaat
      841 atagacaagt tgaaaaattt aataaaaaaa taagtcctca gctcttatat attaagctac
      901 caacttagta tataagccaa aacttaaatg tgctaccaac acatcaagcc gttagagaac
      961 tctatctata gcaatatttc aaatgtaccg acatacaaga gaaacattaa ctatatatat
     1021 tcaatttatg agattatctt aacagatata aatgtaaatt gcaataagta agatttagaa
     1081 gtttatagcc tttgtgtatt ggaagcagta cgcaaaggct tttttatttg ataaaaatta
     1141 gaagtatatt tattttttca taattaattt atgaaaatga aagggggtga gcaaagtgac
     1201 agaggaaagc agtatcttat caaataacaa ggtattagca atatcattat tgactttagc
     1261 agtaaacatt atgactttta tagtgcttgt agctaagtag tacgaaaggg ggagctttaa
     1321 aaagctcctt ggaatacata gaattcataa attaatttat gaaaagaagg gcgtatatga
     1381 aaacttgtaa aaattgcaaa gagtttatta aagatactga aatatgcaaa atacattcgt
     1441 tgatgattca tgataaaaca gtagcaacct attgcagtaa atacaatgag tcaagatgtt
     1501 tacataaagg gaaagtccaa tgtattaatt gttcaaagat gaaccgatat ggatggtgtg
     1561 ccataaaaat gagatgtttt acagaggaag aacagaaaaa agaacgtaca tgcattaaat
     1621 attatgcaag gagctttaaa aaagctcatg taaagaagag taaaaagaaa aaataattta
     1681 tttattaatt taatattgag agtgccgaca cagtatgcac taaaaaatat atctgtggtg
     1741 tagtgagccg atacaaaagg atagtcactc gcattttcat aatacatctt atgttatgat
     1801 tatgtgtcgg tgggacttca cgacgaaaac ccacaataaa aaaagagttc ggggtagggt
     1861 taagcatagt tgaggcaact aaacaatcaa gctaggatat gcagtagcag accgtaaggt
     1921 cgttgtttag gtgtgttgta atacatacgc tattaagatg taaaaatacg gataccaatg
     1981 aagggaaaag tataattttt ggatgtagtt tgtttgttca tctatgggca aactacgtcc
     2041 aaagccgttt ccaaatctgc taaaaagtat atcctttcta aaatcaaagt caagtatgaa
     2101 atcataaata aagtttaatt ttgaagttat tatgatatta tgtttttcta ttaaaataaa
     2161 ttaagtatat agaatagttt aataatagta tatacttaat gtgataagtg tctgacagtg
     2221 tcacagaaag gatgattgtt atggattata agcggccggc cagtgggcaa gttgaaaaat
     2281 tcacaaaaat gtggtataat atctttgttc attagagcga taaacttgaa tttgagaggg
     2341 aacttagatg gtatttgaaa aaattgataa aaatagttgg aacagaaaag agtattttga
     2401 ccactacttt gcaagtgtac cttgtaccta cagcatgacc gttaaagtgg atatcacaca
     2461 aataaaggaa aagggaatga aactatatcc tgcaatgctt tattatattg caatgattgt
     2521 aaaccgccat tcagagttta ggacggcaat caatcaagat ggtgaattgg ggatatatga
     2581 tgagatgata ccaagctata caatatttca caatgatact gaaacatttt ccagcctttg
     2641 gactgagtgt aagtctgact ttaaatcatt tttagcagat tatgaaagtg atacgcaacg
     2701 gtatggaaac aatcatagaa tggaaggaaa gccaaatgct ccggaaaaca tttttaatgt
     2761 atctatgata ccgtggtcaa ccttcgatgg ctttaatctg aatttgcaga aaggatatga
     2821 ttatttgatt cctattttta ctatggggaa atattataaa gaagataaca aaattatact
     2881 tcctttggca attcaagttc atcacgcagt atgtgacgga tttcacattt gccgttttgt
     2941 aaacgaattg caggaattga taaatagtta acttcaggtt tgtctgtaac taaaaacaag
     3001 tatttaagca aaaacatcgt agaaatacgg tgttttttgt taccctaagt ttaaactcct
     3061 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3121 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3181 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3241 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3301 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3361 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3421 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3481 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3541 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3601 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3661 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3721 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3781 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3841 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3901 gagcgaggaa gcggaagagc gcccaatacg cagggccccc tgcttcgggg tcattatagc
     3961 gattttttcg gtatatccat cctttttcgc acgatataca ggattttgcc aaagggttcg
     4021 tgtagacttt ccttggtgta tccaacggcg tcagccgggc aggataggtg aagtaggccc
     4081 acccgcgagc gggtgttcct tcttcactgt cccttattcg cacctggcgg tgctcaacgg
     4141 gaatcctgct ctgcgaggct ggccggctac cgccggcgta acagatgagg gcaagcggat
     4201 ggctgatgaa accaagccaa ccaggaaggg cagcccacct atcaaggtgt actgccttcc
     4261 agacgaacga agagcgattg aggaaaaggc ggcggcggcc ggcatgagcc tgtcggccta
     4321 cctgctggcc gtcggccagg gctacaaaat cacgggcgtc gtggactatg agcacgtccg
     4381 cgagctggcc cgcatcaatg gcgacctggg ccgcctgggc ggcctgctga aactctggct
     4441 caccgacgac ccgcgcacgg cgcggttcgg tgatgccacg atcctcgccc tgctggcgaa
     4501 gatcgaagag aagcaggacg agcttggcaa ggtcatgatg ggcgtggtcc gcccgagggc
     4561 agagccatga cttttttagc cgctaaaacg gccggggggt gcgcgtgatt gccaagcacg
     4621 tccccatgcg ctccatcaag aagagcgact tcgcggagct ggtgaagtac atcaccgacg
     4681 agcaaggcaa gaccgatcgg gccc
//
