LOCUS       V004423                 7598 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004423
VERSION     V004423
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7598)
  AUTHORS   Heap JT, Kuehne SA, Ehsaan M, Cartman ST, Cooksley CM, Scott JC,
            Minton NP.
  TITLE     The ClosTron: Mutagenesis in Clostridium refined and streamlined
  JOURNAL   J. Microbiol. Methods 80 (1), 49-55 (2010)
   PUBMED   19891996
REFERENCE   2  (bases 1 to 7598)
  AUTHORS   Heap JT, Kuehne KA, Ehsaan M, Cartman ST, Cooksley CM, Minton NP.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-SEP-2010) School of Molecular Medical Sciences, The
            University of Nottingham, Centre for Biomolecular Sciences,
            Nottingham, Nottinghamshire NG72RD, UK
REFERENCE   3  (bases 1 to 7598)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7598)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J.
            Microbiol. Methods"; date: "2010"; volume: "80"; issue: "1"; pages:
            "49-55"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (28-SEP-2010) School of Molecular Medical Sciences, The University
            of Nottingham, Centre for Biomolecular Sciences, Nottingham,
            Nottinghamshire NG72RD, UK"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7598
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             238..411
                     /label="lacZ-alpha"
                     /note="LacZ-alpha fragment of beta-galactosidase"
     RBS             1070..1092
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1284..3080
                     /gene="ltrA"
                     /label="Group II intron-encoded protein LtrA"
                     /note="Group II intron-encoded protein LtrA from
                     Lactococcus lactis subsp. cremoris (strain MG1363).
                     Accession#: P0A3U1"
     terminator      3268..3354
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3446..3473
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             5247..5867
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      6047..6635
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     oriT            6956..7065
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             7098..7466
                     /label="traJ"
                     /note="oriT-recognizing protein"
ORIGIN
        1 ggccccctgc agggtgtagt agcctgtgaa ataagtaagg aaaaaaaaga agtaagtgtt
       61 atatatgatg attattttgt agatgtagat aggataatag aatccataga aaatataggt
      121 tatacagtta tataaaaatt actttaaaaa ttaataaaaa catggtaaaa tataaatcgt
      181 ataaagttgt gtaattttta agcttgagct cataacaatt tcacacagga aacagctatg
      241 accatgatta cggattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
      301 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
      361 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgctaa
      421 taaagatctt gtacaatctg taggagaacc tatgggaacg aaacgaaagc gatgccgaga
      481 atctgaattt accaagactt aacactaact ggggataccc taaacaagaa tgcctaatag
      541 aaaggaggaa aaaggctata gcactagagc ttgaaaatct tgcaagggta cggagtactc
      601 gtagtagtct gagaagggta acgcccttta catggcaaag gggtacagtt attgtgtact
      661 aaaattaaaa attgattagg gaggaaaacc tcaaaatgaa accaacaatg gcaattttag
      721 aaagaatcag taaaaattca caagaaaata tagacgaagt ttttacaaga ctttatcgtt
      781 atcttttacg tccagatatt tattacgtgg cgacgcgttg ggaaatggca atgatagcga
      841 aacaacgtaa aactcttgtt gtatgctttc attgtcatcg tcacgtgatt cataaacaca
      901 agtgaatgtc gacagtgaat ttttacgaac gaacaataac agagccgtat actccgagag
      961 gggtacgtac ggttcccgaa gagggtggtg caaaccagtc acagtaatgt gaacaaggcg
     1021 gtacctccct acttcaccat atcattttct gcagccccct agaaataatt ttgtttaact
     1081 ttaagaagga gatatacata tatggctaga tcgtccattc cgacagcatc gccagtcact
     1141 atggcgtgct gctagcgcta tatgcgttga tgcaatttct atgcactcgt agtagtctga
     1201 gaagggtaac gccctttaca tggcaaaggg gtacagttat tgtgtactaa aattaaaaat
     1261 tgattaggga ggaaaacctc aaaatgaaac caacaatggc aattttagaa agaatcagta
     1321 aaaattcaca agaaaatata gacgaagttt ttacaagact ttatcgttat cttttacgtc
     1381 cagatattta ttacgtggcg tatcaaaatt tatattccaa taaaggagct tccacaaaag
     1441 gaatattaga tgatacagcg gatggcttta gtgaagaaaa aataaaaaag attattcaat
     1501 ctttaaaaga cggaacttac tatcctcaac ctgtacgaag aatgtatatt gcaaaaaaga
     1561 attctaaaaa gatgagacct ttaggaattc caactttcac agataaattg atccaagaag
     1621 ctgtgagaat aattcttgaa tctatctatg aaccggtatt cgaagatgtg tctcacggtt
     1681 ttagacctca acgaagctgt cacacagctt tgaaaacaat caaaagagag tttggcggcg
     1741 caagatggtt tgtggaggga gatataaaag gctgcttcga taatatagac cacgttacac
     1801 tcattggact catcaatctt aaaatcaaag atatgaaaat gagccaattg atttataaat
     1861 ttctaaaagc aggttatctg gaaaactggc agtatcacaa aacttacagc ggaacacctc
     1921 aaggtggaat tctatctcct cttttggcca acatctatct tcatgaattg gataagtttg
     1981 ttttacaact caaaatgaag tttgaccgag aaagtccaga aagaataaca cctgaatatc
     2041 gggagctcca caatgagata aaaagaattt ctcaccgtct caagaagttg gagggtgaag
     2101 aaaaagctaa agttctttta gaatatcaag aaaaacgtaa aagattaccc acactcccct
     2161 gtacctcaca gacaaataaa gtattgaaat acgtccggta tgcggacgac ttcattatct
     2221 ctgttaaagg aagcaaagag gactgtcaat ggataaaaga acaattaaaa ctttttattc
     2281 ataacaagct aaaaatggaa ttgagtgaag aaaaaacact catcacacat agcagtcaac
     2341 ccgctcgttt tctgggatat gatatacgag taaggagatc tggaacgata aaacgatctg
     2401 gtaaagtcaa aaagagaaca ctcaatggga gtgtagaact ccttattcct cttcaagaca
     2461 aaattcgtca atttattttt gacaagaaaa tagctatcca aaagaaagat agctcatggt
     2521 ttccagttca caggaaatat cttattcgtt caacagactt agaaatcatc acaatttata
     2581 attctgaact ccgcgggatt tgtaattact acggtctagc aagtaatttt aaccagctca
     2641 attattttgc ttatcttatg gaatacagct gtctaaaaac gatagcctcc aaacataagg
     2701 gaacactttc aaaaaccatt tccatgttta aagatggaag tggttcgtgg gggatcccgt
     2761 atgagataaa gcaaggtaag cagcgccgtt attttgcaaa ttttagtgaa tgtaaatccc
     2821 cttatcaatt tacggatgag ataagtcaag ctcctgtatt gtatggctat gcccggaata
     2881 ctcttgaaaa caggttaaaa gctaaatgtt gtgaattatg tgggacgtct gatgaaaata
     2941 cttcctatga aattcaccat gtcaataagg tcaaaaatct taaaggcaaa gaaaaatggg
     3001 aaatggcaat gatagcgaaa caacgtaaaa ctcttgttgt atgctttcat tgtcatcgtc
     3061 acgtgattca taaacacaag tgaatgtcga gcacccgttc tcggagcact gtccgaccgc
     3121 tttggccgcc gcccagtcct gctcgcttcg ctacttggag ccactatcga ctacgcgatc
     3181 atggcgacca cacccgtcct gtggatcgcc aagccgccga tggtagtgtg gggtctcccc
     3241 atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     3301 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     3361 ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca
     3421 taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt
     3481 ctacaaactc ttcctgtcgt catatctaca agccatcccc ccacagatac gggcgcgccg
     3541 ccattatttt tttgaacaat tgacaattca tttcttattt tttattaagt gatagtcaaa
     3601 aggcataaca gtgctgaata gaaagaaatt tacagaaaag aaaattatag aatttagtat
     3661 gattaattat actcatttat gaatgtttaa ttgaatacaa aaaaaaatac ttgttatgta
     3721 ttcaattacg ggttaaaata tagacaagtt gaaaaattta ataaaaaaat aagtcctcag
     3781 ctcttatata ttaagctacc aacttagtat ataagccaaa acttaaatgt gctaccaaca
     3841 catcaagccg ttagagaact ctatctatag caatatttca aatgtaccga catacaagag
     3901 aaacattaac tatatatatt caatttatga gattatctta acagatataa atgtaaattg
     3961 caataagtaa gatttagaag tttatagcct ttgtgtattg gaagcagtac gcaaaggctt
     4021 ttttatttga taaaaattag aagtatattt attttttcat aattaattta tgaaaatgaa
     4081 agggggtgag caaagtgaca gaggaaagca gtatcttatc aaataacaag gtattagcaa
     4141 tatcattatt gactttagca gtaaacatta tgacttttat agtgcttgta gctaagtagt
     4201 acgaaagggg gagctttaaa aagctccttg gaatacatag aattcataaa ttaatttatg
     4261 aaaagaaggg cgtatatgaa aacttgtaaa aattgcaaag agtttattaa agatactgaa
     4321 atatgcaaaa tacattcgtt gatgattcat gataaaacag tagcaaccta ttgcagtaaa
     4381 tacaatgagt caagatgttt acataaaggg aaagtccaat gtattaattg ttcaaagatg
     4441 aaccgatatg gatggtgtgc cataaaaatg agatgtttta cagaggaaga acagaaaaaa
     4501 gaacgtacat gcattaaata ttatgcaagg agctttaaaa aagctcatgt aaagaagagt
     4561 aaaaagaaaa aataatttat ttattaattt aatattgaga gtgccgacac agtatgcact
     4621 aaaaaatata tctgtggtgt agtgagccga tacaaaagga tagtcactcg cattttcata
     4681 atacatctta tgttatgatt atgtgtcggt gggacttcac gacgaaaacc cacaataaaa
     4741 aaagagttcg gggtagggtt aagcatagtt gaggcaacta aacaatcaag ctaggatatg
     4801 cagtagcaga ccgtaaggtc gttgtttagg tgtgttgtaa tacatacgct attaagatgt
     4861 aaaaatacgg ataccaatga agggaaaagt ataatttttg gatgtagttt gtttgttcat
     4921 ctatgggcaa actacgtcca aagccgtttc caaatctgct aaaaagtata tcctttctaa
     4981 aatcaaagtc aagtatgaaa tcataaataa agtttaattt tgaagttatt atgatattat
     5041 gtttttctat taaaataaat taagtatata gaatagttta ataatagtat atacttaatg
     5101 tgataagtgt ctgacagtgt cacagaaagg atgattgtta tggattataa gcggccggcc
     5161 agtgggcaag ttgaaaaatt cacaaaaatg tggtataata tctttgttca ttagagcgat
     5221 aaacttgaat ttgagaggga acttagatgg tatttgaaaa aattgataaa aatagttgga
     5281 acagaaaaga gtattttgac cactactttg caagtgtacc ttgtacctac agcatgaccg
     5341 ttaaagtgga tatcacacaa ataaaggaaa agggaatgaa actatatcct gcaatgcttt
     5401 attatattgc aatgattgta aaccgccatt cagagtttag gacggcaatc aatcaagatg
     5461 gtgaattggg gatatatgat gagatgatac caagctatac aatatttcac aatgatactg
     5521 aaacattttc cagcctttgg actgagtgta agtctgactt taaatcattt ttagcagatt
     5581 atgaaagtga tacgcaacgg tatggaaaca atcatagaat ggaaggaaag ccaaatgctc
     5641 cggaaaacat ttttaatgta tctatgatac cgtggtcaac cttcgatggc tttaatctga
     5701 atttgcagaa aggatatgat tatttgattc ctatttttac tatggggaaa tattataaag
     5761 aagataacaa aattatactt cctttggcaa ttcaagttca tcacgcagta tgtgacggat
     5821 ttcacatttg ccgttttgta aacgaattgc aggaattgat aaatagttaa cttcaggttt
     5881 gtctgtaact aaaaacaagt atttaagcaa aaacatcgta gaaatacggt gttttttgtt
     5941 accctaagtt taaactcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     6001 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6061 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6121 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6181 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6241 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6301 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6361 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6421 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6481 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6541 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6601 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6661 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6721 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     6781 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc agggccccct
     6841 gcttcggggt cattatagcg attttttcgg tatatccatc ctttttcgca cgatatacag
     6901 gattttgcca aagggttcgt gtagactttc cttggtgtat ccaacggcgt cagccgggca
     6961 ggataggtga agtaggccca cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc
     7021 acctggcggt gctcaacggg aatcctgctc tgcgaggctg gccggctacc gccggcgtaa
     7081 cagatgaggg caagcggatg gctgatgaaa ccaagccaac caggaagggc agcccaccta
     7141 tcaaggtgta ctgccttcca gacgaacgaa gagcgattga ggaaaaggcg gcggcggccg
     7201 gcatgagcct gtcggcctac ctgctggccg tcggccaggg ctacaaaatc acgggcgtcg
     7261 tggactatga gcacgtccgc gagctggccc gcatcaatgg cgacctgggc cgcctgggcg
     7321 gcctgctgaa actctggctc accgacgacc cgcgcacggc gcggttcggt gatgccacga
     7381 tcctcgccct gctggcgaag atcgaagaga agcaggacga gcttggcaag gtcatgatgg
     7441 gcgtggtccg cccgagggca gagccatgac ttttttagcc gctaaaacgg ccggggggtg
     7501 cgcgtgattg ccaagcacgt ccccatgcgc tccatcaaga agagcgactt cgcggagctg
     7561 gtgaagtaca tcaccgacga gcaaggcaag accgatcg
//
