pUC18T-mini-Tn7T-Gm-REP vector (V002415)

Basic Vector Information

      • Vector Name:
      • pUC18T-mini-Tn7T-Gm-REP
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 4967 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Choi KH, Gaynor JB, White KG, Lopez C, Bosio CM, Karkhoff-Schweizer RR, Schweizer HP.

pUC18T-mini-Tn7T-Gm-REP vector Vector Map

pUC18T-mini-Tn7T-Gm-REP4967 bp6001200180024003000360042004800Tn7KS primerlambda t0 terminatorrrnB T1 terminatorFRTGmRPc promoterFRT; Flp recombinase target siteR6K gamma oriTn7LoriToriAmpRAmpR promoter

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

pUC18T-mini-Tn7T-Gm-REP vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_45063        4967 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUC18T-mini-Tn7T-Gm-REP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4967)
  AUTHORS   Choi KH, Gaynor JB, White KG, Lopez C, Bosio CM, Karkhoff-Schweizer 
            RR, Schweizer HP.
  TITLE     A Tn7-based broad-range bacterial cloning and expression system
  JOURNAL   Nat. Methods 2 (6), 443-448 (2005)
  PUBMED    15908923
REFERENCE   2  (bases 1 to 4967)
  AUTHORS   Choi K-H., Gaynor JB, Karkhoff-Schweizer RR, White KG, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-AUG-2004) Microbiology, Immunology and Pathology, 
            Colorado State University, Fort Collins, CO 80523, USA
REFERENCE   3  (bases 1 to 4967)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4967)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Methods"; date: "2005"; volume: "2"; issue: "6"; pages: "443-448"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (09-AUG-2004) Microbiology, Immunology and Pathology, Colorado State
            University, Fort Collins, CO 80523, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4967
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  382..580
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7
                     /note="Tn7R; right end of Tn7"
     primer_bind     585..601
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      629..723
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      826..912
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     protein_bind    958..1005
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(1138..1668)
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     promoter        complement(1857..1885)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     misc_recomb     1926..1973
                     /note="FRT; Flp recombinase target site"
     protein_bind    1926..1973
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     rep_origin      complement(2070..2426)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     mobile_element  complement(2484..2649)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            complement(2808..2916)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(3148..3736)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3910..4767)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4768..4872)
                     /label=AmpR promoter

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