Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V011502 | pGL4.49[luc2P/TCF-LEF RE/Hygro] | 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:
- pGL4.49[luc2P/TCF-LEF RE/Hygro]
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6124 bp
- Type:
- Luciferase Vectors
- Replication origin:
- ori
- Source/Author:
- Promega
- Copy Number:
- High copy number
- Promoter:
- minP
pGL4.49[luc2P/TCF-LEF RE/Hygro] 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.
pGL4.49[luc2P/TCF-LEF RE/Hygro] vector Sequence
LOCUS pGL4.49[luc2P_TC 6124 bp DNA circular SYN 01-JAN-1980
DEFINITION Vector with a minimal promoter and a TCF/LEF response element for
studying cell signaling using destabilized luciferase.
ACCESSION .
VERSION .
KEYWORDS pGL4.49[luc2P TCF-LEF RE Hygro]
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6124)
AUTHORS Promega
TITLE Direct Submission
REFERENCE 2 (bases 1 to 6124)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6124
/mol_type="other DNA"
/organism="synthetic DNA construct"
polyA_signal 105..153
/label=poly(A) signal
/note="synthetic polyadenylation signal"
misc_feature 167..258
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
protein_bind 285..404
/label=TCF/LEF transcription factors binding site
/bound_moiety="TCF/LEF transcription factors"
/note="TCF/LEF RE"
/note="TCF/LEF response element (8 copies)"
promoter 449..480
/label=minP
/note="minimal TATA-box promoter with low basal activity"
CDS 513..2162
/label=luciferase
/note="firefly luciferase"
CDS 2166..2285
/label=hPEST
/note="PEST degradation sequence from mouse ornithine
decarboxylase"
polyA_signal complement(2337..2458)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter 2652..3009
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 3040..4074
/label=HygR
/note="hygromycin B phosphotransferase"
polyA_signal 4101..4149
/label=synthetic polyadenylation signal
/note="synthetic polyadenylation signal"
rep_origin complement(4476..5064)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5267..6124)
/label=AmpR
/note="beta-lactamase"