Basic Vector Information
pUC18R6K-mini-Tn7T-Km vector Vector Map
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 ℃.
pUC18R6K-mini-Tn7T-Km vector Sequence
LOCUS 40924_45038 4466 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pUC18R6K-mini-Tn7T-Km, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4466)
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 4466)
AUTHORS Choi K-H., Gaynor JB, Karkhoff-Schweizer RR, Schweizer HP.
TITLE Direct Submission
JOURNAL Submitted (06-MAY-2004) Microbiology, Immunology and Pathology,
Colorado State University, Fort Collins, CO 80523, USA
REFERENCE 3 (bases 1 to 4466)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4466)
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
(06-MAY-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..4466
/mol_type="other DNA"
/organism="synthetic DNA construct"
repeat_region 382..580
/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"
misc_feature 966..1013
/label=FRT, Flp recombinase target site
/note="FRT, Flp recombinase target site"
protein_bind complement(966..1013)
/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)."
CDS 1163..1975
/label=KanR
/note="aminoglycoside phosphotransferase"
protein_bind complement(2088..2135)
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
misc_feature 2154..2231
/label=MCS, multiple cloning site
/note="MCS, multiple cloning site"
mobile_element complement(2240..2405)
/label=Tn7L
/note="mini-Tn7 element (left end of the Tn7 transposon)"
rep_origin 2641..2997
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
CDS complement(3409..4266)
/label=AmpR
/note="beta-lactamase"
promoter complement(4267..4371)
/label=AmpR promoter
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