Basic Vector Information
- Vector Name:
- pET-21d(+)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5440 bp
- Type:
- pET & Duet Vectors (Novagen)
- Replication origin:
- ori
- Source/Author:
- Novagen (EMD Millipore)
- Copy Number:
- High copy number
pET-21d(+) 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-21d(+) vector Sequence
LOCUS pET-21d(+). 5440 bp DNA circular SYN 01-JAN-1980
DEFINITION Bacterial vector for inducible expression of N-terminally T7-tagged
proteins. Same reading frame as pET-21c(+) but with an NcoI site at
the start codon.
ACCESSION .
VERSION .
KEYWORDS pET-21d(+).
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5440)
AUTHORS Novagen (EMD Millipore)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 5440)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5440
/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(140..157)
/label=6xHis
/note="6xHis affinity tag"
misc_feature 158..203
/label=MCS
/note="MCS"
/note="multiple cloning site"
CDS complement(205..237)
/label=T7 tag (gene 10 leader)
/note="leader peptide from bacteriophage T7 gene 10"
RBS complement(244..266)
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind complement(281..305)
/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(306..324)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 633..710
/label=lacI promoter
CDS 711..1790
/label=lacI
/note="lac repressor"
protein_bind 1806..1827
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS 2602..2790
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
misc_feature 2895..3037
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(3223..3811)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3985..4842)
/label=AmpR
/note="beta-lactamase"
promoter complement(4843..4947)
/label=AmpR promoter
rep_origin complement(4974..5429)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
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