Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V011028 | pET-16b | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pET-16b
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5708 bp
- Type:
- pET & Duet Vectors (Novagen)
- Replication origin:
- ori
- Source/Author:
- Novagen (EMD Millipore)
- Copy Number:
- High copy number
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pET-16b vector Map
Plasmid 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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
References
- Gajanayaka ND, Jo E, Bandara MS, Marasinghe SD, Hettiarachchi SA, Wijewickrama S, Park GH, Oh C, Lee Y. Pseudoalteromonas agarivorans-derived novel ulvan lyase of polysaccharide lyase family 40: Potential application of ulvan and partially hydrolyzed products in cosmetic industry. J Ind Microbiol Biotechnol. 2024 Dec 31;52:kuaf004. doi: 10.1093/jimb/kuaf004. PMID: 39919756; PMCID: PMC11835017.
pET-16b vector Sequence
LOCUS Exported 5708 bp DNA circular SYN 16-SEP-2025
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 5708)
AUTHORS Novagen (EMD Millipore)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 5708)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 5708)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5708)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT SGRef: number: 2; type: "Journal Article"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5708
/mol_type="other DNA"
/organism="synthetic DNA construct"
source join(609..5708,1..608)
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 127..145
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 146..170
/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 185..207
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 214..216
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
CDS 220..246
/label=9xHis
/note="9xHis affinity tag"
CDS 262..273
/label=Factor Xa site
/note="Factor Xa recognition and cleavage site"
misc_feature 274..290
/label=MCS
/note="MCS"
/note="multiple cloning site"
terminator 349..396
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 571..599
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
promoter 712..816
/label=AmpR promoter
CDS 817..1674
/label=AmpR
/note="beta-lactamase"
rep_origin 1848..2436
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 2622..2761
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(2866..3054)
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
protein_bind 4332..4353
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(4369..5448)
/label=lacI
/note="lac repressor"
promoter 5449..5526
/label=lacI promoter