Basic Vector Information
- Vector Name:
- pET-50b(+)
- Antibiotic Resistance:
- Kanamycin
- Length:
- 6733 bp
- Type:
- pET & Duet Vectors (Novagen)
- Replication origin:
- ori
- Source/Author:
- Novagen (EMD Millipore)
- Copy Number:
- High copy number
pET-50b(+) 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 ℃.
pET-50b(+) vector Sequence
LOCUS pET-50b(+). 6733 bp DNA circular SYN 01-JAN-1980 DEFINITION Bacterial vector for expressing proteins with a cleavable N-terminal NusA tag and a cleavable C-terminal S-tag. ACCESSION . VERSION . KEYWORDS pET-50b(+). SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6733) AUTHORS Novagen (EMD Millipore) TITLE Direct Submission REFERENCE 2 (bases 1 to 6733) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..6733 /mol_type="other DNA" /organism="synthetic DNA construct" terminator complement(26..73) /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS complement(168..212) /label=S-Tag /note="affinity and epitope tag derived from pancreatic ribonuclease A" CDS complement(213..230) /label=thrombin site /note="thrombin recognition and cleavage site" misc_feature 234..308 /label=MCS /note="MCS" /note="multiple cloning site" CDS complement(309..332) /label=HRV 3C site /note="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" CDS complement(342..359) /label=6xHis /note="6xHis affinity tag" CDS complement(375..1859) /label=NusA /note="transcription elongation protein (N utilization substance protein A)" CDS complement(1866..1883) /label=6xHis /note="6xHis affinity tag" CDS complement(1893..1895) /codon_start=1 /product="start codon" /label=start codon /note="ATG" /translation="M" RBS complement(1903..1925) /label=RBS /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" protein_bind complement(1940..1964) /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)." promoter complement(1965..1983) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter 2296..2373 /label=lacI promoter CDS 2374..3453 /label=lacI /note="lac repressor" protein_bind 3469..3490 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." CDS 4028..4216 /label=rop /note="Rop protein, which maintains plasmids at low copy number" misc_feature 4321..4463 /label=bom /note="basis of mobility region from pBR322" rep_origin complement(4649..5237) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS 5359..6171 /label=KanR /note="aminoglycoside phosphotransferase" rep_origin complement(6267..6722) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
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