LOCUS       V004425                 8857 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004425
VERSION     V004425
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8857)
  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 8857)
  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 8857)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8857)
  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..8857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             229..402
                     /label="lacZ-alpha"
                     /note="LacZ-alpha fragment of beta-galactosidase"
     CDS             complement(826..1560)
                     /gene="ermBP"
                     /label="rRNA adenine N-6-methyltransferase"
                     /note="rRNA adenine N-6-methyltransferase from Enterococcus
                     faecalis. Accession#: P0A4D5"
     RBS             2320..2342
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2534..4330
                     /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      4518..4604
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4696..4723
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             6497..7117
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      7297..7885
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     oriT            8206..8315
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             8348..8716
                     /label="traJ"
                     /note="oriT-recognizing protein"
ORIGIN
        1 cagggtgtag tagcctgtga aataagtaag gaaaaaaaag aagtaagtgt tatatatgat
       61 gattattttg tagatgtaga taggataata gaatccatag aaaatatagg ttatacagtt
      121 atataaaaat tactttaaaa attaataaaa acatggtaaa atataaatcg tataaagttg
      181 tgtaattttt aagcttgagc tcataacaat ttcacacagg aaacagctat gaccatgatt
      241 acggattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
      301 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
      361 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcta ataaagatct
      421 tgtacaatct gtaggagaac ctatgggaac gaaacgaaag cgatgccgag aatctgaatt
      481 taccaagact taacactaac tggggatacc ctaaacaaga atgcctaata gaaaggagga
      541 aaaaggctat agcactagag cttgaaaatc ttgcaagggt acggagtact cgtagtagtc
      601 tgagaagggt aacgcccttt acatggcaaa ggggtacagt tattgtgtac taaaattaaa
      661 aattgattag ggaggaaaac ctcaaaatga aaccaacaat ggcaatttta gaaagaatca
      721 gtaaaaattc acaagaaaat atagacgaag tttttacaag actttatcgt tatcttttac
      781 gtccagatat ttattacgtg gcgacgcgtg cgactcatag aattatttcc tcccgttaaa
      841 taatagataa ctattaaaaa tagacaatac ttgctcataa gtaacggtac ttaaattgtt
      901 tactttggcg tgtttcattg cttgatgaaa ctgattttta gtaaacagtt gacgatattc
      961 tcgattgacc cattttgaaa caaagtacgt atatagcttc caatatttat ctggaacatc
     1021 tgtggtatgg cgggtaagtt ttattaagac actgtttact tttggtttag gatgaaagca
     1081 ttccgctggc agcttaagca attgctgaat cgagacttga gtgtgcaaga gcaaccctag
     1141 tgttcggtga atatccaagg tacgcttgta gaatccttct tcaacaatca gatagatgtc
     1201 agacgcatgg ctttcaaaaa ccactttttt aataatttgt gtgcttaaat ggtaaggaat
     1261 actcccaaca attttatacc tctgtttgtt agggaattga aactgtagaa tatcttggtg
     1321 aattaaagtg acacgagtat tcagttttaa tttttctgac gataagttga atagatgact
     1381 gtctaattca atagacgtta cctgtttact tattttagcc agtttcgtcg ttaaatgccc
     1441 tttacctgtt ccaatttcgt aaacggtatc ggtttctttt aaattcaatt gttttattat
     1501 ttggttgagt actttttcac tcgttaaaaa gttttgagaa tattttatat ttttgttcat
     1561 accagcacca gaagcaccag catctcttgg gttaattgag gcctgagtat aaggtgactt
     1621 atacttgtaa tctatctaaa cggggaacct ctctagtaga caatcccgtg ctaaattgta
     1681 ggactgccct ttaataaata cttctatatt taaagaggta tttatgaaaa gcggaattta
     1741 tcagattaaa aatactttct ctagagaaaa tttcgtctgg attagttact tatcgtgtaa
     1801 aatctgataa atggaattgg ttctacataa atgcctaacg actatccctt tggggagtag
     1861 ggtcaagtga ctcgaaacga tagacaactt gctttaacaa gttggagata tagtctgctc
     1921 tgcatggtga catgcagctg gatataattc cggggtaaga ttaacgacct tatctgaaca
     1981 taatgccata tgaatccctc ctaatttata cgttttctct aacaacttaa ttatacccac
     2041 tattattatt tttatcaata taacgcgttg ggaaatggca atgatagcga aacaacgtaa
     2101 aactcttgtt gtatgctttc attgtcatcg tcacgtgatt cataaacaca agtgaatgtc
     2161 gacagtgaat ttttacgaac gaacaataac agagccgtat actccgagag gggtacgtac
     2221 ggttcccgaa gagggtggtg caaaccagtc acagtaatgt gaacaaggcg gtacctccct
     2281 acttcaccat atcattttct gcagccccct agaaataatt ttgtttaact ttaagaagga
     2341 gatatacata tatggctaga tcgtccattc cgacagcatc gccagtcact atggcgtgct
     2401 gctagcgcta tatgcgttga tgcaatttct atgcactcgt agtagtctga gaagggtaac
     2461 gccctttaca tggcaaaggg gtacagttat tgtgtactaa aattaaaaat tgattaggga
     2521 ggaaaacctc aaaatgaaac caacaatggc aattttagaa agaatcagta aaaattcaca
     2581 agaaaatata gacgaagttt ttacaagact ttatcgttat cttttacgtc cagatattta
     2641 ttacgtggcg tatcaaaatt tatattccaa taaaggagct tccacaaaag gaatattaga
     2701 tgatacagcg gatggcttta gtgaagaaaa aataaaaaag attattcaat ctttaaaaga
     2761 cggaacttac tatcctcaac ctgtacgaag aatgtatatt gcaaaaaaga attctaaaaa
     2821 gatgagacct ttaggaattc caactttcac agataaattg atccaagaag ctgtgagaat
     2881 aattcttgaa tctatctatg aaccggtatt cgaagatgtg tctcacggtt ttagacctca
     2941 acgaagctgt cacacagctt tgaaaacaat caaaagagag tttggcggcg caagatggtt
     3001 tgtggaggga gatataaaag gctgcttcga taatatagac cacgttacac tcattggact
     3061 catcaatctt aaaatcaaag atatgaaaat gagccaattg atttataaat ttctaaaagc
     3121 aggttatctg gaaaactggc agtatcacaa aacttacagc ggaacacctc aaggtggaat
     3181 tctatctcct cttttggcca acatctatct tcatgaattg gataagtttg ttttacaact
     3241 caaaatgaag tttgaccgag aaagtccaga aagaataaca cctgaatatc gggagctcca
     3301 caatgagata aaaagaattt ctcaccgtct caagaagttg gagggtgaag aaaaagctaa
     3361 agttctttta gaatatcaag aaaaacgtaa aagattaccc acactcccct gtacctcaca
     3421 gacaaataaa gtattgaaat acgtccggta tgcggacgac ttcattatct ctgttaaagg
     3481 aagcaaagag gactgtcaat ggataaaaga acaattaaaa ctttttattc ataacaagct
     3541 aaaaatggaa ttgagtgaag aaaaaacact catcacacat agcagtcaac ccgctcgttt
     3601 tctgggatat gatatacgag taaggagatc tggaacgata aaacgatctg gtaaagtcaa
     3661 aaagagaaca ctcaatggga gtgtagaact ccttattcct cttcaagaca aaattcgtca
     3721 atttattttt gacaagaaaa tagctatcca aaagaaagat agctcatggt ttccagttca
     3781 caggaaatat cttattcgtt caacagactt agaaatcatc acaatttata attctgaact
     3841 ccgcgggatt tgtaattact acggtctagc aagtaatttt aaccagctca attattttgc
     3901 ttatcttatg gaatacagct gtctaaaaac gatagcctcc aaacataagg gaacactttc
     3961 aaaaaccatt tccatgttta aagatggaag tggttcgtgg gggatcccgt atgagataaa
     4021 gcaaggtaag cagcgccgtt attttgcaaa ttttagtgaa tgtaaatccc cttatcaatt
     4081 tacggatgag ataagtcaag ctcctgtatt gtatggctat gcccggaata ctcttgaaaa
     4141 caggttaaaa gctaaatgtt gtgaattatg tgggacgtct gatgaaaata cttcctatga
     4201 aattcaccat gtcaataagg tcaaaaatct taaaggcaaa gaaaaatggg aaatggcaat
     4261 gatagcgaaa caacgtaaaa ctcttgttgt atgctttcat tgtcatcgtc acgtgattca
     4321 taaacacaag tgaatgtcga gcacccgttc tcggagcact gtccgaccgc tttggccgcc
     4381 gcccagtcct gctcgcttcg ctacttggag ccactatcga ctacgcgatc atggcgacca
     4441 cacccgtcct gtggatcgcc aagccgccga tggtagtgtg gggtctcccc atgcgagagt
     4501 agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     4561 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
     4621 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
     4681 ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc
     4741 ttcctgtcgt catatctaca agccatcccc ccacagatac gggcgcgccg ccattatttt
     4801 tttgaacaat tgacaattca tttcttattt tttattaagt gatagtcaaa aggcataaca
     4861 gtgctgaata gaaagaaatt tacagaaaag aaaattatag aatttagtat gattaattat
     4921 actcatttat gaatgtttaa ttgaatacaa aaaaaaatac ttgttatgta ttcaattacg
     4981 ggttaaaata tagacaagtt gaaaaattta ataaaaaaat aagtcctcag ctcttatata
     5041 ttaagctacc aacttagtat ataagccaaa acttaaatgt gctaccaaca catcaagccg
     5101 ttagagaact ctatctatag caatatttca aatgtaccga catacaagag aaacattaac
     5161 tatatatatt caatttatga gattatctta acagatataa atgtaaattg caataagtaa
     5221 gatttagaag tttatagcct ttgtgtattg gaagcagtac gcaaaggctt ttttatttga
     5281 taaaaattag aagtatattt attttttcat aattaattta tgaaaatgaa agggggtgag
     5341 caaagtgaca gaggaaagca gtatcttatc aaataacaag gtattagcaa tatcattatt
     5401 gactttagca gtaaacatta tgacttttat agtgcttgta gctaagtagt acgaaagggg
     5461 gagctttaaa aagctccttg gaatacatag aattcataaa ttaatttatg aaaagaaggg
     5521 cgtatatgaa aacttgtaaa aattgcaaag agtttattaa agatactgaa atatgcaaaa
     5581 tacattcgtt gatgattcat gataaaacag tagcaaccta ttgcagtaaa tacaatgagt
     5641 caagatgttt acataaaggg aaagtccaat gtattaattg ttcaaagatg aaccgatatg
     5701 gatggtgtgc cataaaaatg agatgtttta cagaggaaga acagaaaaaa gaacgtacat
     5761 gcattaaata ttatgcaagg agctttaaaa aagctcatgt aaagaagagt aaaaagaaaa
     5821 aataatttat ttattaattt aatattgaga gtgccgacac agtatgcact aaaaaatata
     5881 tctgtggtgt agtgagccga tacaaaagga tagtcactcg cattttcata atacatctta
     5941 tgttatgatt atgtgtcggt gggacttcac gacgaaaacc cacaataaaa aaagagttcg
     6001 gggtagggtt aagcatagtt gaggcaacta aacaatcaag ctaggatatg cagtagcaga
     6061 ccgtaaggtc gttgtttagg tgtgttgtaa tacatacgct attaagatgt aaaaatacgg
     6121 ataccaatga agggaaaagt ataatttttg gatgtagttt gtttgttcat ctatgggcaa
     6181 actacgtcca aagccgtttc caaatctgct aaaaagtata tcctttctaa aatcaaagtc
     6241 aagtatgaaa tcataaataa agtttaattt tgaagttatt atgatattat gtttttctat
     6301 taaaataaat taagtatata gaatagttta ataatagtat atacttaatg tgataagtgt
     6361 ctgacagtgt cacagaaagg atgattgtta tggattataa gcggccggcc agtgggcaag
     6421 ttgaaaaatt cacaaaaatg tggtataata tctttgttca ttagagcgat aaacttgaat
     6481 ttgagaggga acttagatgg tatttgaaaa aattgataaa aatagttgga acagaaaaga
     6541 gtattttgac cactactttg caagtgtacc ttgtacctac agcatgaccg ttaaagtgga
     6601 tatcacacaa ataaaggaaa agggaatgaa actatatcct gcaatgcttt attatattgc
     6661 aatgattgta aaccgccatt cagagtttag gacggcaatc aatcaagatg gtgaattggg
     6721 gatatatgat gagatgatac caagctatac aatatttcac aatgatactg aaacattttc
     6781 cagcctttgg actgagtgta agtctgactt taaatcattt ttagcagatt atgaaagtga
     6841 tacgcaacgg tatggaaaca atcatagaat ggaaggaaag ccaaatgctc cggaaaacat
     6901 ttttaatgta tctatgatac cgtggtcaac cttcgatggc tttaatctga atttgcagaa
     6961 aggatatgat tatttgattc ctatttttac tatggggaaa tattataaag aagataacaa
     7021 aattatactt cctttggcaa ttcaagttca tcacgcagta tgtgacggat ttcacatttg
     7081 ccgttttgta aacgaattgc aggaattgat aaatagttaa cttcaggttt gtctgtaact
     7141 aaaaacaagt atttaagcaa aaacatcgta gaaatacggt gttttttgtt accctaagtt
     7201 taaactcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     7261 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     7321 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     7381 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     7441 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     7501 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7561 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7621 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     7681 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7741 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     7801 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7861 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7921 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     7981 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     8041 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc agggccccct gcttcggggt
     8101 cattatagcg attttttcgg tatatccatc ctttttcgca cgatatacag gattttgcca
     8161 aagggttcgt gtagactttc cttggtgtat ccaacggcgt cagccgggca ggataggtga
     8221 agtaggccca cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc acctggcggt
     8281 gctcaacggg aatcctgctc tgcgaggctg gccggctacc gccggcgtaa cagatgaggg
     8341 caagcggatg gctgatgaaa ccaagccaac caggaagggc agcccaccta tcaaggtgta
     8401 ctgccttcca gacgaacgaa gagcgattga ggaaaaggcg gcggcggccg gcatgagcct
     8461 gtcggcctac ctgctggccg tcggccaggg ctacaaaatc acgggcgtcg tggactatga
     8521 gcacgtccgc gagctggccc gcatcaatgg cgacctgggc cgcctgggcg gcctgctgaa
     8581 actctggctc accgacgacc cgcgcacggc gcggttcggt gatgccacga tcctcgccct
     8641 gctggcgaag atcgaagaga agcaggacga gcttggcaag gtcatgatgg gcgtggtccg
     8701 cccgagggca gagccatgac ttttttagcc gctaaaacgg ccggggggtg cgcgtgattg
     8761 ccaagcacgt ccccatgcgc tccatcaaga agagcgactt cgcggagctg gtgaagtaca
     8821 tcaccgacga gcaaggcaag accgatcggg ccccctg
//
