Basic Vector Information
- Vector Name:
- pET-28a-BFP-C
- Antibiotic Resistance:
- Kanamycin
- Length:
- 6064 bp
- Type:
- E. coli Expression Vectors, Fluorescent Protein Re
- Replication origin:
- ori
- Selection Marker:
- TagBFP
- Promoter:
- T7/lac
pET-28a-BFP-C 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-28a-BFP-C vector Sequence
LOCUS 40924_17806 6064 bp DNA circular SYN 13-JAN-2022 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6064) TITLE Direct Submission REFERENCE 2 (bases 1 to 6064) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..6064 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin 12..467 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" CDS complement(563..1375) /label=KanR /note="aminoglycoside phosphotransferase" rep_origin 1497..2085 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature complement(2271..2413) /label=bom /note="basis of mobility region from pBR322" CDS complement(2518..2706) /label=rop /note="Rop protein, which maintains plasmids at low copy number" protein_bind complement(3481..3502) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." CDS complement(3518..4597) /label=lacI /note="lac repressor" promoter complement(4598..4675) /label=lacI promoter promoter 4984..5002 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 5003..5027 /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 5042..5064 /label=RBS /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 5083..5100 /label=6xHis /note="6xHis affinity tag" CDS 5110..5127 /label=thrombin site /note="thrombin recognition and cleavage site" CDS 5131..5163 /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=leader peptide from bacteriophage T7 gene 10 /note="T7 tag (gene 10 leader)" /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" CDS 5200..5898 /label=TagBFP /note="monomeric blue fluorescent protein" CDS 5908..5925 /label=6xHis /note="6xHis affinity tag" terminator 5992..6039 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase"
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