Basic Vector Information
pET-16b 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-16b vector Sequence
LOCUS pET-16b. 5711 bp DNA circular SYN 01-JAN-1980
DEFINITION Bacterial vector for inducible expression of N-terminally
10xHis-tagged proteins with a Factor Xa site.
ACCESSION .
VERSION .
KEYWORDS pET-16b.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5711)
AUTHORS Novagen (EMD Millipore)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 5711)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5711
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 10..38
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
terminator complement(213..260)
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
misc_feature 319..335
/label=MCS
/note="MCS"
/note="multiple cloning site"
CDS complement(336..347)
/label=Factor Xa site
/note="Factor Xa recognition and cleavage site"
CDS complement(363..389)
/label=9xHis
/note="9xHis affinity tag"
CDS complement(393..395)
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
RBS complement(402..424)
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind complement(439..463)
/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(464..482)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 791..868
/label=lacI promoter
CDS 869..1948
/label=lacI
/note="lac repressor"
protein_bind 1964..1985
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS 3263..3451
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
misc_feature 3556..3698
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(3884..4472)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4646..5503)
/label=AmpR
/note="beta-lactamase"
promoter complement(5504..5608)
/label=AmpR promoter
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