Basic Vector Information
pTrcHis B 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 ℃.
pTrcHis B vector Sequence
LOCUS pTrcHis_B. 4404 bp DNA circular SYN 01-JAN-1980 DEFINITION Bacterial vector for inducible expression of N-terminally 6xHis-tagged proteins. For other reading frames, use pTrcHis A or pTrcHis C. ACCESSION . VERSION . KEYWORDS pTrcHis B. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4404) AUTHORS Invitrogen (Life Technologies) TITLE Direct Submission REFERENCE 2 (bases 1 to 4404) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..4404 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 193..222 /label=trc promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 230..246 /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)." misc_feature 263..332 /label=rrnG antiterminator /note="antiterminator from the E. coli rrnG leader region (Berg et al., 1989)" misc_feature 346..354 /label=gene 10 translational enhancer /note="gene 10 translational enhancer" /note="translational enhancer from gene 10 of bacteriophage T7 (Olins and Rangwala, 1989)" RBS 369..373 /note="ribosome binding site" CDS 383..406 /label=minicistron /note="synthetic cistron containing a ribosome binding site (Shine-Dalgarno sequence), for enhancing the bacterial expression of a downstream cistron (Schoner, 1997)" CDS 413..415 /codon_start=1 /product="start codon" /label=start codon /note="ATG" /translation="M" CDS 425..442 /label=6xHis /note="6xHis affinity tag" CDS 446..478 /label=T7 tag (gene 10 leader) /note="leader peptide from bacteriophage T7 gene 10" CDS 482..505 /label=Xpress(TM) tag /note="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" misc_feature 505..554 /label=MCS /note="MCS" /note="multiple cloning site" terminator 757..843 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 935..962 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 982..1073 /label=AmpR promoter CDS 1074..1931 /label=AmpR /note="beta-lactamase" rep_origin 2105..2693 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature complement(2879..3019) /label=bom /note="basis of mobility region from pBR322" promoter 3205..3282 /label=lacIq promoter /note="In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold." CDS 3283..4362 /label=lacI /note="lac repressor"
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