Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V011447 | pRL-CMV | 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
The pRL-CMV is a mammalian co-reporter vector for the strong and constitutive expression of wild-type Renilla luciferase.
- Vector Name:
- pRL-CMV
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4079 bp
- Type:
- Luciferase Vectors
- Replication origin:
- ori
- Source/Author:
- Promega
- Copy Number:
- High copy number
- Promoter:
- CMV
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pRL-CMV 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
- Ieki T, Yamanaka Y, Yoshikawa K. Functional analysis of human olfactory receptors with a high basal activity using LNCaP cell line. PLoS One. 2022 Apr 21;17(4):e0267356. doi: 10.1371/journal.pone.0267356. PMID: 35446888; PMCID: PMC9022881.
pRL-CMV vector Sequence
LOCUS 40924_37188 4079 bp DNA circular SYN 18-DEC-2018
DEFINITION Co-reporter vector pRL-CMV, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4079)
AUTHORS Sherf B, Navarro S, Wood KV.
TITLE Dual-Luciferase(TM) Reporter Assay: An Advanced Co-reporter
Technology
JOURNAL Promega Notes 57, 2-5 (1996)
REFERENCE 2 (bases 1 to 4079)
AUTHORS Sherf B, Navarro S, Wood KV.
TITLE Direct Submission
JOURNAL Submitted (20-SEP-1997) Production, Promega Co., 5445 East Cheryl
Parkway, Madison, WI 53711, USA
REFERENCE 3 (bases 1 to 4079)
AUTHORS Grooms K.
TITLE Direct Submission
JOURNAL Submitted (17-DEC-1999) Technical Writing, Promega Co., 2800 Woods
Hollow Rd., Madison, WI 53711, USA
REFERENCE 4 (bases 1 to 4079)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 4079)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Promega
Notes 57, 2-5 (1996)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(20-SEP-1997) Production, Promega Co., 5445 East Cheryl Parkway,
Madison, WI 53711, USA"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(17-DEC-1999) Technical Writing, Promega Co., 2800 Woods Hollow Rd.,
Madison, WI 53711, USA"
COMMENT SGRef: number: 4; type: "Journal Article"
COMMENT On Dec 17, 1999 this sequence version replaced AF025843.1.
FEATURES Location/Qualifiers
source 1..4079
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 145..523
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 524..727
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
intron 863..995
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
promoter 1040..1058
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1068..2000
/codon_start=1
/label=Rluc
/note="luciferase from the anthozoan coelenterate Renilla
reniformis (sea pansy)"
/translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
polyA_signal complement(2034..2155)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter 2288..2392
/label=AmpR promoter
CDS 2393..3250
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 3424..4012
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"