LOCUS       Exported                9196 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  ClosTron mutagenesis vector pMTL007S-E2, complete sequence.
ACCESSION   HQ327468
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9196)
  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 9196)
  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 9196)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9196
                     /organism="ClosTron mutagenesis vector pMTL007S-E2"
                     /mol_type="other DNA"
                     /db_xref="taxon:907254"
     primer_bind     221..237
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             233..406
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     primer_bind     complement(253..269)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(814..847)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    complement(2101..2134)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     RBS             2392..2414
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      4590..4676
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4768..4795
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      7640..8228
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     oriT            8549..8658
                     /note="incP origin of transfer"
     CDS             8691..9062
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"