Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012411 | pQE 80L Kan T7WT | 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
- Vector Name:
- pQE 80L Kan T7WT
- Antibiotic Resistance:
- Kanamycin
- Length:
- 7154 bp
- Type:
- Bacterial Expression
- Replication origin:
- ori
pQE 80L Kan T7WT 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.
pQE 80L Kan T7WT vector Sequence
LOCUS V012411 7154 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V012411
VERSION V012411
KEYWORDS pQE 80L Kan T7WT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 7154)
TITLE Compartmentalized Partnered Replication for the Directed Evolution
of Genetic Parts and Circuits
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7154)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7154)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7154
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter complement(1007..1084)
/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 1276..1416
/label="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(1602..2190)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2300..3112)
/label="KanR"
/note="aminoglycoside phosphotransferase"
promoter 3226..3270
/label="T5 promoter"
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
CDS 3343..3360
/label="6xHis"
/note="6xHis affinity tag"
CDS 3367..6012
/note="T7 RNA polymerase from Escherichia phage T7.
Accession#: P00573"
/label="T7 RNA polymerase"
terminator 6037..6131
/label="lambda t0 terminator"
/note="transcription terminator from phage lambda"
CDS 6175..6831
/label="CmR"
/note="chloramphenicol acetyltransferase"
terminator 6899..6985
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
protein_bind complement(7044..7065)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(join(7081..7154,1..1006))
/label="lacI"
/note="lac repressor"