Basic Vector Information
pUC18T-mini-Tn7T-Gm-REP 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 ℃.
pUC18T-mini-Tn7T-Gm-REP vector Sequence
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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