LOCUS       V004429                 5028 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004429
VERSION     V004429
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5028)
  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 5028)
  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 5028)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5028)
  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..5028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      9..62
                     /label="derived from CD0164 gene of Clostridium difficile"
                     /note="derived from CD0164 gene of Clostridium difficile"
                     /regulatory_class="terminator"
     primer_bind     63..79
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    90..389
                     /gene="thl fragment"
                     /note="mediates homologous recombination with the
                     chromosome of Clostridium acetobutylicum ATCC824"
     gene            90..389
                     /gene="thl fragment"
                     /label="thl fragment"
     CDS             412..987
                     /codon_start=1
                     /gene="pyrE"
                     /product="orotate phosphoribosyltransferase"
                     /label="pyrE"
                     /protein_id="AFA34996.1"
                     /translation="MSNINVIDILKESNALLEGHFLLSSGRHSNRYCQCAKLLQYPQKA
                     EKVISVVAEKLKDADFNIIVGPAMGGVIVSYELARQTNRPGIFAERKDGIMCIRRGFEI
                     KKGDKVIISEDVITTGKSSLEVAKVIEEMGGEVVGIACIVDRRAEGIKTNYPIYSACKL
                     EIETYEKDNCELCKKNIPFVKPGSREQK"
     gene            412..987
                     /gene="pyrE"
                     /label="pyrE"
     primer_bind     complement(1106..1122)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    1270..2469
                     /note="mediates homologous recombination with the
                     chromosome of Clostridium acetobutylicum ATCC824"
     CDS             1705..2070
                     /codon_start=1
                     /gene="CAC2872"
                     /product="putative FoF1-type ATP synthase operon membrane
                     protein"
                     /label="CAC2872"
                     /protein_id="AFA34998.1"
                     /translation="MNLNIKRMLKVVTLYDAIIAAIVSVILLFAANYKISLIVIIGIFS
                     AIFNFYLSNLTADFVFVKKMGNTSLIFLSSIFRVILVFFIGIILYKIYKYYLIAYLGGY
                     SAHFIALIIYGSLVNKR"
     gene            1705..2070
                     /gene="CAC2872"
                     /label="CAC2872"
     CDS             2848..3288
                     /codon_start=1
                     /gene="repL"
                     /product="plasmid replication protein"
                     /label="repL"
                     /protein_id="AFA34999.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     gene            2848..3288
                     /gene="repL"
                     /label="repL"
     CDS             3438..4058
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      4238..4826
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 cctgcaggat aaaaaaattg tagataaatt ttataaaata gttttatcta caattttttt
       61 atcaggaaac agctatgacc gcggccatct atgcaacaaa agcagctatt gaaaaagcag
      121 gttggacagt tgatgaatta gatttaatag aatcaaatga agcttttgca gctcaaagtt
      181 tagcagtagc aaaagattta aaatttgata tgaataaagt aaatgtaaat ggaggagcta
      241 ttgcccttgg tcatccaatt ggagcatcag gtgcaagaat actcgttact cttgtacacg
      301 caatgcaaaa aagagatgca aaaaaaggct tagcaacttt atgtataggt ggcggacaag
      361 gaacagcaat attgctagaa aagtgctaga tcgattaagg agaagatata aatgagtaat
      421 ataaatgtta tagatatatt aaaagaatca aatgcattat tagaagggca tttcttacta
      481 tcatctggaa gacatagtaa tagatattgt caatgtgcga aactacttca atatccacaa
      541 aaagcagaaa aagttattag tgtagtagca gaaaagctta aagatgcaga ctttaatata
      601 atagtaggac cagctatggg tggtgtaata gtatcttatg aattagcaag gcaaactaat
      661 agaccaggca tatttgcaga aagaaaagat ggaattatgt gtatacgaag aggatttgaa
      721 ataaaaaaag gagataaagt tataatatca gaggatgtaa taactacagg aaaatcttct
      781 ttagaagtag ctaaagttat agaagaaatg ggtggagaag tagtaggtat agcatgcata
      841 gttgatagaa gagcggaagg tataaaaaca aactacccaa tatatagtgc ttgtaaatta
      901 gaaatagaaa cttatgaaaa agataattgt gagttatgta aaaaaaatat accttttgta
      961 aaaccaggta gtagagaaca aaaataagcg gccgctgtat ccatatgacc atgattacga
     1021 attcgagctc ggtacccggg gatcctctag agtcgacgtc acgcgtccat ggagatctcg
     1081 aggcctgcag acatgcaagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     1141 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     1201 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     1261 ggcgctagca aagtattgtt aaaaataact ctgtagaatt ataaattagt tctacagagt
     1321 tattttttaa aaaaattcta aacttatgta taaaaaatac gataagaatg tagaattaaa
     1381 actaaagaca gttcaatttc ttttagaata atttagttag tgtggtaaaa aaatgtcata
     1441 atgatattta tgttgaaatt tgtataaaat tcagaaaatg aatatatttt atcaattttc
     1501 agtcatttga aagattatga ggctaatgca gtactaggcg taaattgaat ttataattac
     1561 tatagcgata agaaatggcc taaaaacgtt tgcagtaatg aaagaaccgt aaatattata
     1621 aaaaaaatct taaaacagag ttttatttat aaaaatttaa gatatataat ttaaataacg
     1681 tgttaaaata gtggaggaag taatttgaat ctgaatatta aaagaatgtt aaaggttgta
     1741 actctttatg atgcaattat tgctgcaata gtttcagtaa tacttttgtt tgctgctaat
     1801 tataagattt cgttaatagt gattataggg attttttcag caatatttaa tttttattta
     1861 agtaatttaa cagctgattt cgtttttgta aaaaaaatgg gaaatacgtc acttatattt
     1921 cttagttcaa tttttagagt aatacttgtt ttttttatag gtattattct ttataaaata
     1981 tataaatatt atttaatagc ctacttagga ggatatagtg ctcattttat agcccttata
     2041 atttatgggt cactagtaaa taaacgatga aaggaagtga ttgaatggag ctaggtgcaa
     2101 agacagtatt ttcgatgaag cttggaagtt acaactttgc tataacagaa actgtagtat
     2161 tacagtggat tatcatggca gttataatat tacttgcaat atttcttact aaaaatctta
     2221 agaaagtacc aaataggaaa caaagcgtaa tagaaatgat tgttaactta ataaatggat
     2281 tggtaaaaga aaatatggga gagaaattca tgaatttcgt tccaattatc ggtactatgg
     2341 cagtgtttat acttttctta aatttaacag ggctagtagg tatcgaacca gcaacaaagg
     2401 atattagtgt tacagcaggc tttgctttag taagtgcatt tttaataaat gcaactgcaa
     2461 taaaaagaag gcgcgccgca ttcacttctt ttctatataa atatgagcga agcgaataag
     2521 cgtcggaaaa gcagcaaaaa gtttcctttt tgctgttgga gcatgggggt tcagggggtg
     2581 cagtatctga cgtcaatgcc gagcgaaagc gagccgaagg gtagcattta cgttagataa
     2641 ccccctgata tgctccgacg ctttatatag aaaagaagat tcaactaggt aaaatcttaa
     2701 tataggttga gatgataagg tttataagga atttgtttgt tctaattttt cactcatttt
     2761 gttctaattt cttttaacaa atgttctttt ttttttagaa cagttatgat atagttagaa
     2821 tagtttaaaa taaggagtga gaaaaagatg aaagaaagat atggaacagt ctataaaggc
     2881 tctcagaggc tcatagacga agaaagtgga gaagtcatag aggtagacaa gttataccgt
     2941 aaacaaacgt ctggtaactt cgtaaaggca tatatagtgc aattaataag tatgttagat
     3001 atgattggcg gaaaaaaact taaaatcgtt aactatatcc tagataatgt ccacttaagt
     3061 aacaatacaa tgatagctac aacaagagaa atagcaaaag ctacaggaac aagtctacaa
     3121 acagtaataa caacacttaa aatcttagaa gaaggaaata ttataaaaag aaaaactgga
     3181 gtattaatgt taaaccctga actactaatg agaggcgacg accaaaaaca aaaatacctc
     3241 ttactcgaat ttgggaactt tgagcaagag gcaaatgaaa tagattgacc tcccaataac
     3301 accacgtagt tattgggagg tcaatctatg aaatgcgatt aagggccggc cagtgggcaa
     3361 gttgaaaaat tcacaaaaat gtggtataat atctttgttc attagagcga taaacttgaa
     3421 tttgagaggg aacttagatg gtatttgaaa aaattgataa aaatagttgg aacagaaaag
     3481 agtattttga ccactacttt gcaagtgtac cttgtaccta cagcatgacc gttaaagtgg
     3541 atatcacaca aataaaggaa aagggaatga aactatatcc tgcaatgctt tattatattg
     3601 caatgattgt aaaccgccat tcagagttta ggacggcaat caatcaagat ggtgaattgg
     3661 ggatatatga tgagatgata ccaagctata caatatttca caatgatact gaaacatttt
     3721 ccagcctttg gactgagtgt aagtctgact ttaaatcatt tttagcagat tatgaaagtg
     3781 atacgcaacg gtatggaaac aatcatagaa tggaaggaaa gccaaatgct ccggaaaaca
     3841 tttttaatgt atctatgata ccgtggtcaa ccttcgatgg ctttaatctg aatttgcaga
     3901 aaggatatga ttatttgatt cctattttta ctatggggaa atattataaa gaagataaca
     3961 aaattatact tcctttggca attcaagttc atcacgcagt atgtgacgga tttcacattt
     4021 gccgttttgt aaacgaattg caggaattga taaatagtta acttcaggtt tgtctgtaac
     4081 taaaaacaag tatttaagca aaaacatcgt agaaatacgg tgttttttgt taccctaagt
     4141 ttaaactcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4201 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4261 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4321 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4381 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4441 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4501 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4561 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4621 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4681 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4741 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4801 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4861 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4921 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     4981 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg cagggccc
//
