Basic Vector Information
p15TV-L 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 ℃.
p15TV-L vector Sequence
LOCUS 40924_2428 7746 bp DNA circular SYN 18-DEC-2018 DEFINITION Expression vector p15TV-L, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7746) AUTHORS Guthrie J, Loppnau P, Kozieradzki I, Savchenko A, Arrowsmith C. TITLE Expression vectors for high-throughput In-Fusion cloning JOURNAL Unpublished REFERENCE 2 (bases 1 to 7746) AUTHORS Guthrie J, Loppnau P, Kozieradzki I, Savchenko A, Arrowsmith C. TITLE Direct Submission JOURNAL Submitted (26-FEB-2007) Structural Genomics Consortium, University of Toronto, 100 College Street, Toronto, ON M5G1L5, Canada REFERENCE 3 (bases 1 to 7746) TITLE Direct Submission REFERENCE 4 (bases 1 to 7746) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (26-FEB-2007) Structural Genomics Consortium, University of Toronto, 100 College Street, Toronto, ON M5G1L5, Canada" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..7746 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 104..208 /label=AmpR promoter CDS 209..1066 /label=AmpR /note="beta-lactamase" rep_origin 1240..1828 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature complement(2014..2156) /label=bom /note="basis of mobility region from pBR322" CDS complement(2261..2449) /label=rop /note="Rop protein, which maintains plasmids at low copy number" protein_bind complement(3727..3748) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." CDS complement(3764..4843) /label=lacI /note="lac repressor" promoter complement(4844..4921) /label=lacI promoter promoter 5230..5248 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 5249..5273 /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 5288..5310 /label=RBS /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 5329..5346 /label=6xHis /note="6xHis affinity tag" CDS 5359..5379 /label=TEV site /note="tobacco etch virus (TEV) protease recognition and cleavage site" protein_bind complement(5407..5440) /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." promoter 5455..5900 /label=sacB promoter /note="sacB promoter and control region" CDS 5901..7319 /label=SacB /note="secreted levansucrase that renders bacterial growth sensitive to sucrose" misc_feature 7395..7409 /note="3'-LIC sequence; add complement to downstream PCR primer" terminator 7487..7534 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter complement(7709..7737) /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene"
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