LOCUS       V004421                 9128 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004421
VERSION     V004421
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9128)
  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 9128)
  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 9128)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9128)
  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..9128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             243..416
                     /label="lacZ-alpha"
                     /note="LacZ-alpha fragment of beta-galactosidase"
     CDS             complement(840..1574)
                     /gene="ermBP"
                     /label="rRNA adenine N-6-methyltransferase"
                     /note="rRNA adenine N-6-methyltransferase from Enterococcus
                     faecalis. Accession#: P0A4D5"
     RBS             2334..2356
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2548..4344
                     /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      4532..4618
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4710..4737
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     rep_origin      7582..8170
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     oriT            8491..8600
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             8633..9001
                     /label="traJ"
                     /note="oriT-recognizing protein"
ORIGIN
        1 gatcgggccc cctgcagggt gtagtagcct gtgaaataag taaggaaaaa aaagaagtaa
       61 gtgttatata tgatgattat tttgtagatg tagataggat aatagaatcc atagaaaata
      121 taggttatac agttatataa aaattacttt aaaaattaat aaaaacatgg taaaatataa
      181 atcgtataaa gttgtgtaat ttttaagctt gagctcataa caatttcaca caggaaacag
      241 ctatgaccat gattacggat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
      301 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
      361 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
      421 gctaataaag atcttgtaca atctgtagga gaacctatgg gaacgaaacg aaagcgatgc
      481 cgagaatctg aatttaccaa gacttaacac taactgggga taccctaaac aagaatgcct
      541 aatagaaagg aggaaaaagg ctatagcact agagcttgaa aatcttgcaa gggtacggag
      601 tactcgtagt agtctgagaa gggtaacgcc ctttacatgg caaaggggta cagttattgt
      661 gtactaaaat taaaaattga ttagggagga aaacctcaaa atgaaaccaa caatggcaat
      721 tttagaaaga atcagtaaaa attcacaaga aaatatagac gaagttttta caagacttta
      781 tcgttatctt ttacgtccag atatttatta cgtggcgacg cgtgcgactc atagaattat
      841 ttcctcccgt taaataatag ataactatta aaaatagaca atacttgctc ataagtaacg
      901 gtacttaaat tgtttacttt ggcgtgtttc attgcttgat gaaactgatt tttagtaaac
      961 agttgacgat attctcgatt gacccatttt gaaacaaagt acgtatatag cttccaatat
     1021 ttatctggaa catctgtggt atggcgggta agttttatta agacactgtt tacttttggt
     1081 ttaggatgaa agcattccgc tggcagctta agcaattgct gaatcgagac ttgagtgtgc
     1141 aagagcaacc ctagtgttcg gtgaatatcc aaggtacgct tgtagaatcc ttcttcaaca
     1201 atcagataga tgtcagacgc atggctttca aaaaccactt ttttaataat ttgtgtgctt
     1261 aaatggtaag gaatactccc aacaatttta tacctctgtt tgttagggaa ttgaaactgt
     1321 agaatatctt ggtgaattaa agtgacacga gtattcagtt ttaatttttc tgacgataag
     1381 ttgaatagat gactgtctaa ttcaatagac gttacctgtt tacttatttt agccagtttc
     1441 gtcgttaaat gccctttacc tgttccaatt tcgtaaacgg tatcggtttc ttttaaattc
     1501 aattgtttta ttatttggtt gagtactttt tcactcgtta aaaagttttg agaatatttt
     1561 atatttttgt tcataccagc accagaagca ccagcatctc ttgggttaat tgaggcctga
     1621 gtataaggtg acttatactt gtaatctatc taaacgggga acctctctag tagacaatcc
     1681 cgtgctaaat tgtaggactg ccctttaata aatacttcta tatttaaaga ggtatttatg
     1741 aaaagcggaa tttatcagat taaaaatact ttctctagag aaaatttcgt ctggattagt
     1801 tacttatcgt gtaaaatctg ataaatggaa ttggttctac ataaatgcct aacgactatc
     1861 cctttgggga gtagggtcaa gtgactcgaa acgatagaca acttgcttta acaagttgga
     1921 gatatagtct gctctgcatg gtgacatgca gctggatata attccggggt aagattaacg
     1981 accttatctg aacataatgc catatgaatc cctcctaatt tatacgtttt ctctaacaac
     2041 ttaattatac ccactattat tatttttatc aatataacgc gttgggaaat ggcaatgata
     2101 gcgaaacaac gtaaaactct tgttgtatgc tttcattgtc atcgtcacgt gattcataaa
     2161 cacaagtgaa tgtcgacagt gaatttttac gaacgaacaa taacagagcc gtatactccg
     2221 agaggggtac gtacggttcc cgaagagggt ggtgcaaacc agtcacagta atgtgaacaa
     2281 ggcggtacct ccctacttca ccatatcatt ttctgcagcc ccctagaaat aattttgttt
     2341 aactttaaga aggagatata catatatggc tagatcgtcc attccgacag catcgccagt
     2401 cactatggcg tgctgctagc gctatatgcg ttgatgcaat ttctatgcac tcgtagtagt
     2461 ctgagaaggg taacgccctt tacatggcaa aggggtacag ttattgtgta ctaaaattaa
     2521 aaattgatta gggaggaaaa cctcaaaatg aaaccaacaa tggcaatttt agaaagaatc
     2581 agtaaaaatt cacaagaaaa tatagacgaa gtttttacaa gactttatcg ttatctttta
     2641 cgtccagata tttattacgt ggcgtatcaa aatttatatt ccaataaagg agcttccaca
     2701 aaaggaatat tagatgatac agcggatggc tttagtgaag aaaaaataaa aaagattatt
     2761 caatctttaa aagacggaac ttactatcct caacctgtac gaagaatgta tattgcaaaa
     2821 aagaattcta aaaagatgag acctttagga attccaactt tcacagataa attgatccaa
     2881 gaagctgtga gaataattct tgaatctatc tatgaaccgg tattcgaaga tgtgtctcac
     2941 ggttttagac ctcaacgaag ctgtcacaca gctttgaaaa caatcaaaag agagtttggc
     3001 ggcgcaagat ggtttgtgga gggagatata aaaggctgct tcgataatat agaccacgtt
     3061 acactcattg gactcatcaa tcttaaaatc aaagatatga aaatgagcca attgatttat
     3121 aaatttctaa aagcaggtta tctggaaaac tggcagtatc acaaaactta cagcggaaca
     3181 cctcaaggtg gaattctatc tcctcttttg gccaacatct atcttcatga attggataag
     3241 tttgttttac aactcaaaat gaagtttgac cgagaaagtc cagaaagaat aacacctgaa
     3301 tatcgggagc tccacaatga gataaaaaga atttctcacc gtctcaagaa gttggagggt
     3361 gaagaaaaag ctaaagttct tttagaatat caagaaaaac gtaaaagatt acccacactc
     3421 ccctgtacct cacagacaaa taaagtattg aaatacgtcc ggtatgcgga cgacttcatt
     3481 atctctgtta aaggaagcaa agaggactgt caatggataa aagaacaatt aaaacttttt
     3541 attcataaca agctaaaaat ggaattgagt gaagaaaaaa cactcatcac acatagcagt
     3601 caacccgctc gttttctggg atatgatata cgagtaagga gatctggaac gataaaacga
     3661 tctggtaaag tcaaaaagag aacactcaat gggagtgtag aactccttat tcctcttcaa
     3721 gacaaaattc gtcaatttat ttttgacaag aaaatagcta tccaaaagaa agatagctca
     3781 tggtttccag ttcacaggaa atatcttatt cgttcaacag acttagaaat catcacaatt
     3841 tataattctg aactccgcgg gatttgtaat tactacggtc tagcaagtaa ttttaaccag
     3901 ctcaattatt ttgcttatct tatggaatac agctgtctaa aaacgatagc ctccaaacat
     3961 aagggaacac tttcaaaaac catttccatg tttaaagatg gaagtggttc gtgggggatc
     4021 ccgtatgaga taaagcaagg taagcagcgc cgttattttg caaattttag tgaatgtaaa
     4081 tccccttatc aatttacgga tgagataagt caagctcctg tattgtatgg ctatgcccgg
     4141 aatactcttg aaaacaggtt aaaagctaaa tgttgtgaat tatgtgggac gtctgatgaa
     4201 aatacttcct atgaaattca ccatgtcaat aaggtcaaaa atcttaaagg caaagaaaaa
     4261 tgggaaatgg caatgatagc gaaacaacgt aaaactcttg ttgtatgctt tcattgtcat
     4321 cgtcacgtga ttcataaaca caagtgaatg tcgagcaccc gttctcggag cactgtccga
     4381 ccgctttggc cgccgcccag tcctgctcgc ttcgctactt ggagccacta tcgactacgc
     4441 gatcatggcg accacacccg tcctgtggat cgccaagccg ccgatggtag tgtggggtct
     4501 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     4561 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     4621 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
     4681 gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc
     4741 gtttctacaa actcttcctg tcgtcatatc tacaagccat ccccccacag atacgggcgc
     4801 gccgccatta tttttttgaa caattgacaa ttcatttctt attttttatt aagtgatagt
     4861 caaaaggcat aacagtgctg aatagaaaga aatttacaga aaagaaaatt atagaattta
     4921 gtatgattaa ttatactcat ttatgaatgt ttaattgaat acaaaaaaaa atacttgtta
     4981 tgtattcaat tacgggttaa aatatagaca agttgaaaaa tttaataaaa aaataagtcc
     5041 tcagctctta tatattaagc taccaactta gtatataagc caaaacttaa atgtgctacc
     5101 aacacatcaa gccgttagag aactctatct atagcaatat ttcaaatgta ccgacataca
     5161 agagaaacat taactatata tattcaattt atgagattat cttaacagat ataaatgtaa
     5221 attgcaataa gtaagattta gaagtttata gcctttgtgt attggaagca gtacgcaaag
     5281 gcttttttat ttgataaaaa ttagaagtat atttattttt tcataattaa tttatgaaaa
     5341 tgaaaggggg tgagcaaagt gacagaggaa agcagtatct tatcaaataa caaggtatta
     5401 gcaatatcat tattgacttt agcagtaaac attatgactt ttatagtgct tgtagctaag
     5461 tagtacgaaa gggggagctt taaaaagctc cttggaatac atagaattca taaattaatt
     5521 tatgaaaaga agggcgtata tgaaaacttg taaaaattgc aaagagttta ttaaagatac
     5581 tgaaatatgc aaaatacatt cgttgatgat tcatgataaa acagtagcaa cctattgcag
     5641 taaatacaat gagtcaagat gtttacataa agggaaagtc caatgtatta attgttcaaa
     5701 gatgaaccga tatggatggt gtgccataaa aatgagatgt tttacagagg aagaacagaa
     5761 aaaagaacgt acatgcatta aatattatgc aaggagcttt aaaaaagctc atgtaaagaa
     5821 gagtaaaaag aaaaaataat ttatttatta atttaatatt gagagtgccg acacagtatg
     5881 cactaaaaaa tatatctgtg gtgtagtgag ccgatacaaa aggatagtca ctcgcatttt
     5941 cataatacat cttatgttat gattatgtgt cggtgggact tcacgacgaa aacccacaat
     6001 aaaaaaagag ttcggggtag ggttaagcat agttgaggca actaaacaat caagctagga
     6061 tatgcagtag cagaccgtaa ggtcgttgtt taggtgtgtt gtaatacata cgctattaag
     6121 atgtaaaaat acggatacca atgaagggaa aagtataatt tttggatgta gtttgtttgt
     6181 tcatctatgg gcaaactacg tccaaagccg tttccaaatc tgctaaaaag tatatccttt
     6241 ctaaaatcaa agtcaagtat gaaatcataa ataaagttta attttgaagt tattatgata
     6301 ttatgttttt ctattaaaat aaattaagta tatagaatag tttaataata gtatatactt
     6361 aatgtgataa gtgtctgaca gtgtcacaga aaggatgatt gttatggatt ataagcggcc
     6421 ggcccaatga ataggtttac acttacttta gttttatgga aatgaaagat catatcatat
     6481 ataatctaga ataaaattaa ctaaaataat tattatctag ataaaaaatt tagaagccaa
     6541 tgaaatctat aaataaacta aattaagttt atttaattaa caactatgga tataaaatag
     6601 gtactaatca aaatagtgag gaggatatat ttgaatacat acgaacaaat taataaagtg
     6661 aaaaaaatac ttcggaaaca tttaaaaaat aaccttattg gtacttacat gtttggatca
     6721 ggagttgaga gtggactaaa accaaatagt gatcttgact ttttagtcgt cgtatctgaa
     6781 ccattgacag atcaaagtaa agaaatactt atacaaaaaa ttagacctat ttcaaagaaa
     6841 ataggagata aaagcaactt acgatatatt gaattaacaa ttattattca gcaagaaatg
     6901 gtaccgtgga atcatcctcc caaacaagaa tttatttatg gagaatggtt acaagagctt
     6961 tatgaacaag gatacattcc tcagaaggaa ttaaattcag atttaaccat aatgctttac
     7021 caagcaaaac gaaaaaataa aagaatatac ggaaattatg acttagagga attactacct
     7081 gatattccat tttctgatgt gagaagagcc attatggatt cgtcagagga attaatagat
     7141 aattatcagg atgatgaaac caactctata ttaactttat gccgtatgat tttaactatg
     7201 gacacgggta aaatcatacc aaaagatatt gcgggaaatg cagtggctga atcttctcca
     7261 ttagaacata gggagagaat tttgttagca gttcgtagtt atcttggaga gaatattgaa
     7321 tggactaatg aaaatgtaaa tttaactata aactatttaa ataacagatt aaaaaaatta
     7381 taaaaaaatt gaaaaaatgg tggaaacact tttttcaatt tttttgtttt attatttaat
     7441 atttgggaaa tattcattct aattggtaat cagattttag aagtttaaac tcctttttga
     7501 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7561 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7621 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7681 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     7741 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7801 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7861 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7921 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7981 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     8041 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     8101 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     8161 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     8221 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     8281 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     8341 ggaagcggaa gagcgcccaa tacgcagggc cccctgcttc ggggtcatta tagcgatttt
     8401 ttcggtatat ccatcctttt tcgcacgata tacaggattt tgccaaaggg ttcgtgtaga
     8461 ctttccttgg tgtatccaac ggcgtcagcc gggcaggata ggtgaagtag gcccacccgc
     8521 gagcgggtgt tccttcttca ctgtccctta ttcgcacctg gcggtgctca acgggaatcc
     8581 tgctctgcga ggctggccgg ctaccgccgg cgtaacagat gagggcaagc ggatggctga
     8641 tgaaaccaag ccaaccagga agggcagccc acctatcaag gtgtactgcc ttccagacga
     8701 acgaagagcg attgaggaaa aggcggcggc ggccggcatg agcctgtcgg cctacctgct
     8761 ggccgtcggc cagggctaca aaatcacggg cgtcgtggac tatgagcacg tccgcgagct
     8821 ggcccgcatc aatggcgacc tgggccgcct gggcggcctg ctgaaactct ggctcaccga
     8881 cgacccgcgc acggcgcggt tcggtgatgc cacgatcctc gccctgctgg cgaagatcga
     8941 agagaagcag gacgagcttg gcaaggtcat gatgggcgtg gtccgcccga gggcagagcc
     9001 atgacttttt tagccgctaa aacggccggg gggtgcgcgt gattgccaag cacgtcccca
     9061 tgcgctccat caagaagagc gacttcgcgg agctggtgaa gtacatcacc gacgagcaag
     9121 gcaagacc
//
