pMTL007 vector (V004426)

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LOCUS       V004426                11845 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004426
VERSION     V004426
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11845)
  AUTHORS   Heap JT, Pennington OJ, Cartman ST, Carter GP, Minton NP.
  TITLE     The ClosTron: a universal gene knock-out system for the genus
            Clostridium
  JOURNAL   J. Microbiol. Methods 70 (3), 452-464 (2007)
   PUBMED   17658189
REFERENCE   2  (bases 1 to 11845)
  AUTHORS   Heap JT, Pennington OJ, Cartman ST, Carter GP, Minton NP.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-MAR-2007) Centre for Biomolecular Sciences, The
            University of Nottingham, University Park, Nottingham NG7 2RD,
            United Kingdom
REFERENCE   3  (bases 1 to 11845)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 11845)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J.
            Microbiol. Methods"; date: "2007"; volume: "70"; issue: "3"; pages:
            "452-464"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (27-MAR-2007) Centre for Biomolecular Sciences, The University of
            Nottingham, University Park, Nottingham NG7 2RD, United Kingdom"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..11845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..338
                     /note="group II intron IBS and EBS; replaced to alter the
                     intron target specificity"
     CDS             complement(743..1477)
                     /gene="ermBP"
                     /label="rRNA adenine N-6-methyltransferase"
                     /note="rRNA adenine N-6-methyltransferase from Enterococcus
                     faecalis. Accession#: P0A4D5"
     intron          1510..1902
                     /gene="ErmBtdRAM2"
                     /note="group I intron from td gene of phage T4 with
                     intron-encoded protein deletion; inserted in reverse
                     orientation to confer RAM functionality"
     RBS             2237..2259
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2451..4247
                     /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      4436..4522
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4614..4641
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     primer_bind     complement(4888..4904)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             5784..6101
                     /codon_start=1
                     /product="putative plasmid replication protein"
                     /label="putative plasmid replication protein"
                     /note="ORFH"
                     /protein_id="ABU90834.1"
                     /translation="MKRRAYMKTCKNCKEFIKDTEICKIHSLMIHDKTVATYCSKYNES
                     RCLHKGKVQCINCSKMNRYGWCAIKMRCFTEEEQKKERTCIKYYARSFKKAHVKKSKKK
                     K"
     CDS             6893..7513
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      7998..8586
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8880..9959)
                     /label="lacI"
                     /note="lac repressor"
     oriT            10671..10780
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             10813..11181
                     /label="traJ"
                     /note="oriT-recognizing protein"
     regulatory      11585..11686
                     /label="IPTG-inducible promoter fac"
                     /note="IPTG-inducible promoter fac"
                     /regulatory_class="promoter"
     protein_bind    complement(11692..11716)
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             11735..11743
                     /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)"