Basic Vector Information
- Vector Name:
- pTrcHis2 A
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4406 bp
- Type:
- Cloning Vectors
- Replication origin:
- ori
- Source/Author:
- Invitrogen (Life Technologies)
- Copy Number:
- High copy number
pTrcHis2 A 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 ℃.
pTrcHis2 A vector Sequence
LOCUS pTrcHis2_A. 4406 bp DNA circular SYN 01-JAN-1980 DEFINITION Bacterial vector for inducible expression of C-terminally 6xHis-tagged proteins. For other reading frames, use pTrcHis2 B or pTrcHis2 C. ACCESSION . VERSION . KEYWORDS pTrcHis2 A. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4406) AUTHORS Invitrogen (Life Technologies) TITLE Direct Submission REFERENCE 2 (bases 1 to 4406) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..4406 /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)" misc_feature 411..464 /label=MCS /note="MCS" /note="multiple cloning site" CDS 413..415 /codon_start=1 /product="start codon" /label=start codon /note="ATG" /translation="M" CDS 471..500 /label=Myc /note="Myc (human c-Myc proto-oncogene) epitope tag" CDS 516..533 /label=6xHis /note="6xHis affinity tag" terminator 759..845 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 937..964 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 984..1075 /label=AmpR promoter CDS 1076..1933 /label=AmpR /note="beta-lactamase" rep_origin 2107..2695 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature complement(2881..3021) /label=bom /note="basis of mobility region from pBR322" promoter 3207..3284 /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 3285..4364 /label=lacI /note="lac repressor"
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