Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V011445 | pRL-SV40 | 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:
- pRL-SV40
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3705 bp
- Type:
- Luciferase Vectors
- Replication origin:
- ori
- Source/Author:
- Promega
- Copy Number:
- High copy number
pRL-SV40 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.
pRL-SV40 vector Sequence
LOCUS 40924_37198 3705 bp DNA circular SYN 18-DEC-2018 DEFINITION Co-reporter vector pRL-SV40, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3705) 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 3705) 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 3705) AUTHORS Sherf B, Navarro S, Wood KV. TITLE Direct Submission JOURNAL Submitted (18-FEB-2000) Production, Promega Co., 5445 East Cheryl Parkway, Madison, WI 53711, USA REFERENCE 4 (bases 1 to 3705) TITLE Direct Submission REFERENCE 5 (bases 1 to 3705) 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 (18-FEB-2000) Production, Promega Co., 5445 East Cheryl Parkway, Madison, WI 53711, USA" COMMENT SGRef: number: 4; type: "Journal Article" COMMENT On Feb 18, 2000 this sequence version replaced AF025845.1. FEATURES Location/Qualifiers source 1..3705 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 62..419 /label=SV40 promoter /note="SV40 enhancer and early promoter" intron 489..621 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" promoter 666..684 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 694..1626 /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(1660..1781) /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" promoter 1914..2018 /label=AmpR promoter CDS 2019..2876 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 3050..3638 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"