LOCUS       V004432                 5388 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004432
VERSION     V004432
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5388)
  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 5388)
  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 5388)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5388)
  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..5388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            9..581
                     /gene="pyrE fragment"
                     /label="pyrE fragment"
     misc_feature    9..308
                     /gene="pyrE fragment"
                     /note="mediates homologous recombination with the
                     chromosome of Clostridium sporogenes NCIMB10696"
     primer_bind     complement(700..716)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    864..2063
                     /note="mediates homologous recombination with the
                     chromosome of Clostridium sporogenes NCIMB10696"
     CDS             1082..1894
                     /codon_start=1
                     /product="hypothetical protein"
                     /label="hypothetical protein"
                     /note="CBO3234-like"
                     /protein_id="AFA34988.1"
                     /translation="MAKPSIFSKDYENRMKKRKRRTFFSVIVIIVIFLGIIFTNNGIGK
                     KIKNSLNQIKEETKAEEEQKNKQQEQKENKKENQNKNAATVKKESEVPKDNNIEVQLEG
                     NIKIKLIYIEDSNKNKTIKSIDLNKNNLLYDINPSKNSIVVTNPKTQNIYLVNLNGEKQ
                     DITNKQYTSTSGTTFQKDAILASKQDYLWGVLPKFIDNDNIAYVSQLPWFNKTTKYVWM
                     YNIKNKTHLYNENISGEDIKFDKLTEKGLTVIADGKTLFLKADGSIAQ"
     RBS             1986..1994
                     /label="Shine-Dalgarno sequence"
                     /note="full consensus sequence for ribosome-binding sites
                     upstream of start codons in E. coli; complementary to a
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             2442..2882
                     /codon_start=1
                     /gene="repL"
                     /product="plasmid replication protein"
                     /label="repL"
                     /protein_id="AFA34989.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     gene            2442..2882
                     /gene="repL"
                     /label="repL"
     CDS             3032..3652
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      3832..4420
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     oriT            4741..4850
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             4883..5251
                     /label="traJ"
                     /note="oriT-recognizing protein"
ORIGIN
        1 cctgcaggag taatataaat gttatagata tattaaaaga atcaaatgca ttattagaag
       61 gtcatttctt actatcatct ggaagacata gtaatagata ttgtcaatgt gcgaaactac
      121 ttcaatatcc acaaaaagca gaaaaagtta ttagtgtaat agcacagaag cttaaaaatg
      181 tagactttaa tataatagta ggaccagcta tgggtggtgt aatagtatcc tatgaattag
      241 caaggcaaac taatagacca ggcatatttg cagaaagaaa agatggaatc atgtgtatac
      301 gaagaggatt tgaaataaaa aaaggagata aagttataat atcagaggat gtaataacta
      361 caggtaaatc ttctttagaa gtagctaaag ttatagaaga aatgggtgga gaagtagtag
      421 gtatagcatg tatagttgat agaagagcgg aaggtataaa aacaaactat caaatatata
      481 gtgcttgtaa attagaaata gaaacttatg aaaaagataa ctgtgagtta tgtaaaaaaa
      541 atataccttt tgtaaaacca ggtagtagag aacaaaaata agcggccgct gtatccatat
      601 gaccatgatt acgaattcga gctcggtacc cggggatcct ctagagtcga cgtcacgcgt
      661 ccatggagat ctcgaggcct gcagacatgc aagcttggca ctggccgtcg ttttacaacg
      721 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
      781 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
      841 cctgaatggc gaatggcgct agcatttaaa aataaggagt gtctcaaaat agatttaatt
      901 ttgagacatt tttttagttt taacaaatat taaacaaaaa cttaatttta ctttaataat
      961 tatgaaaatg gttattaatt tttattgttt tgagttataa tagtgttaat tatatattat
     1021 aataattgat acattttatt tattttagta tagataggta ttatatatag gaggaattat
     1081 tgtggcaaaa ccaagtattt ttagcaaaga ttatgaaaat agaatgaaaa agagaaaaag
     1141 aaggacattt ttttcagtta tagttataat agtaattttt ttaggaataa tattcactaa
     1201 taatggtatt ggtaaaaaaa taaaaaatag tttaaatcaa ataaaagaag aaactaaggc
     1261 tgaagaagaa caaaaaaata aacaacaaga acaaaaggaa aataaaaaag aaaatcaaaa
     1321 taaaaatgca gccacagtaa aaaaagaaag tgaagttcct aaagataata atatagaagt
     1381 tcaattagaa ggcaatataa aaataaagtt gatatatata gaagatagta ataaaaataa
     1441 gactattaaa tctatagatc taaataaaaa taatttatta tatgatataa atccttcaaa
     1501 aaattcaata gttgtaacaa atcctaaaac tcaaaatatt tatttagtaa atttaaatgg
     1561 tgaaaagcaa gatataacaa ataagcagta tacatcaaca tctggaacaa cttttcaaaa
     1621 ggatgctata ttagcgtcaa aacaagatta tttatgggga gttttaccta agttcataga
     1681 taatgataat atagcctatg tttctcaact gccatggttt aataaaacca ctaaatatgt
     1741 atggatgtat aatataaaaa ataaaactca tttatataat gaaaatatat caggagaaga
     1801 cattaagttt gataaattaa cagaaaaagg tcttacagta attgcagatg gcaaaacttt
     1861 atttttaaaa gcagatggaa gtatagctca gtaaattaaa aaatattaaa tataaaagtt
     1921 aataagtagc aattaaaata taaaattttt aagagtgtat gaaatatata cactcttgct
     1981 ttatttaagg aggtaaatat ggaaatagga atgtcttctg cttgtttgta tcctaaagta
     2041 ccaatagaag aaagtatatc agtggcgcgc cgcattcact tcttttctat ataaatatga
     2101 gcgaagcgaa taagcgtcgg aaaagcagca aaaagtttcc tttttgctgt tggagcatgg
     2161 gggttcaggg ggtgcagtat ctgacgtcaa tgccgagcga aagcgagccg aagggtagca
     2221 tttacgttag ataaccccct gatatgctcc gacgctttat atagaaaaga agattcaact
     2281 aggtaaaatc ttaatatagg ttgagatgat aaggtttata aggaatttgt ttgttctaat
     2341 ttttcactca ttttgttcta atttctttta acaaatgttc tttttttttt agaacagtta
     2401 tgatatagtt agaatagttt aaaataagga gtgagaaaaa gatgaaagaa agatatggaa
     2461 cagtctataa aggctctcag aggctcatag acgaagaaag tggagaagtc atagaggtag
     2521 acaagttata ccgtaaacaa acgtctggta acttcgtaaa ggcatatata gtgcaattaa
     2581 taagtatgtt agatatgatt ggcggaaaaa aacttaaaat cgttaactat atcctagata
     2641 atgtccactt aagtaacaat acaatgatag ctacaacaag agaaatagca aaagctacag
     2701 gaacaagtct acaaacagta ataacaacac ttaaaatctt agaagaagga aatattataa
     2761 aaagaaaaac tggagtatta atgttaaacc ctgaactact aatgagaggc gacgaccaaa
     2821 aacaaaaata cctcttactc gaatttggga actttgagca agaggcaaat gaaatagatt
     2881 gacctcccaa taacaccacg tagttattgg gaggtcaatc tatgaaatgc gattaagggc
     2941 cggccagtgg gcaagttgaa aaattcacaa aaatgtggta taatatcttt gttcattaga
     3001 gcgataaact tgaatttgag agggaactta gatggtattt gaaaaaattg ataaaaatag
     3061 ttggaacaga aaagagtatt ttgaccacta ctttgcaagt gtaccttgta cctacagcat
     3121 gaccgttaaa gtggatatca cacaaataaa ggaaaaggga atgaaactat atcctgcaat
     3181 gctttattat attgcaatga ttgtaaaccg ccattcagag tttaggacgg caatcaatca
     3241 agatggtgaa ttggggatat atgatgagat gataccaagc tatacaatat ttcacaatga
     3301 tactgaaaca ttttccagcc tttggactga gtgtaagtct gactttaaat catttttagc
     3361 agattatgaa agtgatacgc aacggtatgg aaacaatcat agaatggaag gaaagccaaa
     3421 tgctccggaa aacattttta atgtatctat gataccgtgg tcaaccttcg atggctttaa
     3481 tctgaatttg cagaaaggat atgattattt gattcctatt tttactatgg ggaaatatta
     3541 taaagaagat aacaaaatta tacttccttt ggcaattcaa gttcatcacg cagtatgtga
     3601 cggatttcac atttgccgtt ttgtaaacga attgcaggaa ttgataaata gttaacttca
     3661 ggtttgtctg taactaaaaa caagtattta agcaaaaaca tcgtagaaat acggtgtttt
     3721 ttgttaccct aagtttaaac tcctttttga taatctcatg accaaaatcc cttaacgtga
     3781 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3841 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3901 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3961 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4021 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4081 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4141 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4201 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4261 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4321 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4381 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4441 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4501 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     4561 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcagggc
     4621 cccctgcttc ggggtcatta tagcgatttt ttcggtatat ccatcctttt tcgcacgata
     4681 tacaggattt tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac ggcgtcagcc
     4741 gggcaggata ggtgaagtag gcccacccgc gagcgggtgt tccttcttca ctgtccctta
     4801 ttcgcacctg gcggtgctca acgggaatcc tgctctgcga ggctggccgg ctaccgccgg
     4861 cgtaacagat gagggcaagc ggatggctga tgaaaccaag ccaaccagga agggcagccc
     4921 acctatcaag gtgtactgcc ttccagacga acgaagagcg attgaggaaa aggcggcggc
     4981 ggccggcatg agcctgtcgg cctacctgct ggccgtcggc cagggctaca aaatcacggg
     5041 cgtcgtggac tatgagcacg tccgcgagct ggcccgcatc aatggcgacc tgggccgcct
     5101 gggcggcctg ctgaaactct ggctcaccga cgacccgcgc acggcgcggt tcggtgatgc
     5161 cacgatcctc gccctgctgg cgaagatcga agagaagcag gacgagcttg gcaaggtcat
     5221 gatgggcgtg gtccgcccga gggcagagcc atgacttttt tagccgctaa aacggccggg
     5281 gggtgcgcgt gattgccaag cacgtcccca tgcgctccat caagaagagc gacttcgcgg
     5341 agctggtgaa gtacatcacc gacgagcaag gcaagaccga tcgggccc
//
