LOCUS       Exported                4941 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMTL83353, complete sequence.
ACCESSION   FJ797648
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4941)
  AUTHORS   Heap JT, Pennington OJ, Cartman ST, Minton NP.
  TITLE     A modular system for Clostridium shuttle plasmids
  JOURNAL   J. Microbiol. Methods 78 (1), 79-85 (2009)
  PUBMED    19445976
REFERENCE   2  (bases 1 to 4941)
  AUTHORS   Pennington OJ, Heap JT, Cartman ST, Minton NP.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-MAR-2009) Centre for Biomolecular Sciences, The 
            University of Nottingham, University Park, Nottingham NG7 2RD, 
            United Kingdom
REFERENCE   3  (bases 1 to 4941)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4941
                     /organism="Shuttle vector pMTL83353"
                     /mol_type="other DNA"
                     /db_xref="taxon:647631"
     regulatory      9..62
                     /regulatory_class="terminator"
                     /note="terminator derived from CD0164 gene of Clostridium 
                     difficile"
     primer_bind     63..80
                     /label=M13R
                     /note="M13R"
     primer_bind     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      89..288
                     /regulatory_class="promoter"
                     /note="promoter derived from fdx gene of Clostridium 
                     sporogenes"
     RBS             274..282
                     /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)"
     misc_feature    292..555
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             292..555
                     /codon_start=1
                     /gene="lacZ alpha"
                     /product="beta-galactosidase, alpha subunit"
                     /label=lacZ alpha
                     /protein_id="ACR43902.1"
                     /translation="MTMITNSSSVPGDPLESTSRVHGDLEACRHASLALAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWR"
     gene            292..555
                     /gene="lacZ alpha"
                     /label=lacZ alpha
     primer_bind     complement(393..410)
                     /label=M13F
                     /note="M13F"
     primer_bind     complement(393..409)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      557..598
                     /regulatory_class="terminator"
                     /note="terminator derived from fdx gene of Clostridium 
                     pasteurianum"
     CDS             1325..1642
                     /codon_start=1
                     /gene="repH"
                     /product="putative plasmid replication protein"
                     /label=repH
                     /protein_id="ACR43901.1"
                     /translation="MKRRAYMKTCKNCKEFIKDTEICKIHSLMIHDKTVATYCSKYNES
                     RCLHKGKVQCINCSKMNRYGWCAIKMRCFTEEEQKKERTCIKYYARSFKKAHVKKSKKK
                     K"
     gene            1325..1642
                     /gene="repH"
                     /label=repH
     CDS             2434..3186
                     /codon_start=1
                     /gene="aad9"
                     /product="spectinomycin resistance protein"
                     /label=aad9
                     /protein_id="ACR43899.1"
                     /translation="MNTYEQINKVKKILRKHLKNNLIGTYMFGSGVESGLKPNSDLDFL
                     VVVSEPLTDQSKEILIQKIRPISKKIGDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYG
                     EWLQELYEQGYIPQKELNSDLTIMLYQAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMD
                     SSEELIDNYQDDETNSILTLCRMILTMDTGKIIPKDIAGNAVAESSPLEHRERILLAVR
                     SYLGENIEWTNENVNLTINYLNNRLKKL"
     gene            2434..3186
                     /gene="aad9"
                     /label=aad9
     rep_origin      3385..3973
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      3421..3966
                     /label=ColE1 RNA II
                     /note="ColE1 RNA II"
     oriT            4294..4403
                     /note="incP origin of transfer"
     CDS             4367..4807
                     /codon_start=1
                     /gene="traJ"
                     /product="plasmid conjugative transfer protein"
                     /label=traJ
                     /protein_id="ACR43900.1"
                     /translation="MLNGNPALRGWPATAGVTDEGKRMADETKPTRKGSPPIKVYCLPD
                     ERRAIEEKAAAAGMSLSAYLLAVGQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWL
                     TDDPRTARFGDATILALLAKIEEKQDELGKVMMGVVRPRAEP"
     gene            4367..4807
                     /gene="traJ"
                     /label=traJ
     CDS             4436..4807
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"