Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010778 | pQE-80L-Kan | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
This is a bacterial lacIq vector with a kanamycin resistance marker for expressing N-terminally 6 x His-tagged proteins.
- Vector Name:
- pQE-80L-Kan
- Antibiotic Resistance:
- Kanamycin, Chloramphenicol
- Length:
- 4541 bp
- Type:
- Qiagen Vectors
- Replication origin:
- ori
- Source/Author:
- Qiagen
- Copy Number:
- High copy number
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pQE-80L-Kan 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
- Smith AC, Cronan JE. Evidence against translational repression by the carboxyltransferase component of Escherichia coli acetyl coenzyme A carboxylase. J Bacteriol. 2014 Nov;196(21):3768-75.
pQE-80L-Kan vector Sequence
LOCUS pQE-80L-Kan. 4541 bp DNA circular SYN 01-JAN-1980
DEFINITION Bacterial lacIq vector with a kanamycin resistance marker for
expressing N-terminally 6xHis-tagged proteins. For other reading
frames, use pQE-81L-Kan or pQE-82L-Kan.
ACCESSION .
VERSION .
KEYWORDS pQE-80L-Kan
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4541)
AUTHORS Qiagen
TITLE Direct Submission
REFERENCE 2 (bases 1 to 4541)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT Because this vector contains the lacIq gene, the pREP4 plasmid is
not needed.
FEATURES Location/Qualifiers
source 1..4541
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 10..54
/label=T5 promoter
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 62..78
/label=lac repressor encoded by lacI binding site
/bound_moiety="lac repressor encoded by lacI"
/note="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 101..106
/note="ribosome binding site"
CDS 115..117
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
CDS 127..144
/label=6xHis
/note="6xHis affinity tag"
misc_feature 145..192
/label=MCS
/note="MCS"
/note="multiple cloning site"
misc_feature 192..202
/label=stop codons
/note="stop codons"
/note="stop codons in all three reading frames"
terminator 208..302
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS 346..1002
/label=CmR
/note="chloramphenicol acetyltransferase"
terminator 1070..1156
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
protein_bind complement(1215..1236)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(1252..2331)
/label=lacI
/note="lac repressor"
promoter complement(2332..2409)
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
misc_feature 2601..2741
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(2927..3515)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3625..4437)
/label=KanR
/note="aminoglycoside phosphotransferase"