Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V017015 | pTetR-LasR-pLuxR-GFP | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pTetR-LasR-pLuxR-GFP
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3977 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- E. coli
- Promoter:
- pLux
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pTetR-LasR-pLuxR-GFP vector 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.
pTetR-LasR-pLuxR-GFP vector Sequence
LOCUS V017015 3977 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V017015
VERSION V017015
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 3977)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..3977
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 83..154
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 170..197
/label="T7Te terminator"
/note="phage T7 early transcription terminator"
misc_feature 204..224
/label="BioBrick suffix"
/note="universal suffix for all parts"
rep_origin complement(412..1000)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1174..2031)
/label="AmpR"
/note="beta-lactamase"
promoter complement(2032..2136)
/label="AmpR promoter"
terminator complement(2226..2269)
/label="bacterial terminator"
/note="putative bacterial transcription terminator"
misc_feature 2272..2293
/label="BioBrick prefix"
/note="BioBrick prefix for parts that do not start with
'ATG'"
protein_bind 2294..2312
/label="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
CDS 2374..3090
/gene="lasR"
/label="Transcriptional activator protein LasR"
/note="Transcriptional activator protein LasR from
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP
104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1).
Accession#: P25084"
terminator complement(3116..3159)
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS join(3324..3977,1..60)
/label="yeGFP"
/note="yeast-enhanced green fluorescent protein"