pRL-SV40 vector (V011445)

COA reports Sequencing Results MSDS Download GenBank File(.gb)

Price Information

Cat No.	Plasmid Name	Availability	Buy one, get one free! (?)
V011445	pRL-SV40	In stock, 1 week for quality controls

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

Copy Sequence View 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"