p2luc vector (V009766)

Price Information

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V009766	p2luc	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

p2luc is a dual-luciferase reporter system for studying recoding signals.

Vector Name:: p2luc

Antibiotic Resistance:: Ampicillin

Length:: 5393 bp

Type:: Reporter vector

Replication origin:: ori

Source/Author:: Grentzmann G, Ingram JA, Kelly PJ, Gesteland RF, Atkins JF.

Copy Number:: High Copy

Promoter:: SV40 promoter

Cloning Method:: Restriction Enzyme

Fusion Tag:: luciferace

Expression Method:: Transient

p2luc vector Vector Map

View Map in NovoBuilder

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

Grentzmann G, Ingram JA, Kelly PJ, Gesteland RF, Atkins JF. A dual-luciferase reporter system for studying recoding signals. RNA. 1998 Apr;4(4):479-86. PMID: 9630253; PMCID: PMC1369633.

p2luc vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       p2luc.        5393 bp DNA     circular SYN 18-FEB-2022
DEFINITION  Reporter vector p2luc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    p2luc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5393)
  AUTHORS   Grentzmann G, Ingram JA, Kelly PJ, Gesteland RF, Atkins JF.
  TITLE     A dual-luciferase reporter system for studying recoding signals
  JOURNAL   RNA 4 (4), 479-486 (1998)
  PUBMED    9630253
REFERENCE   2  (bases 1 to 5393)
  AUTHORS   Grentzmann G, Gesteland RF, Atkins JF.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-JAN-1998) HHMI, University of Utah, 6100 Eccles Bldg.,
            Salt Lake City, UT 84112, USA
REFERENCE   3  (bases 1 to 5393)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5393)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "RNA"; date:
            "1998"; volume: "4"; issue: "4"; pages: "479-486"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-JAN-1998) HHMI, University of Utah, 6100 Eccles Bldg., Salt Lake
            City, UT 84112, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5393
                     /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
                     /label=Rluc
                     /note="luciferase from the anthozoan coelenterate Renilla 
                     reniformis (sea pansy)"
     CDS             1656..3305
                     /label=luciferase
                     /note="firefly luciferase"
     polyA_signal    complement(3348..3469)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        3602..3706
                     /label=AmpR promoter
     CDS             3707..4564
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      4738..5326
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"