Skip to main content
Addgene
Open Menu

pcDNA3 RLUC POLIRES FLUC
(Plasmid #45642)

Print

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 45642 Standard format: Plasmid sent in bacteria as agar stab 1 $94
Add to Cart
DNA 45642-D50 50 μg of DNA in Tris buffer $495
Add to Cart
DNA 45642-D100 100 μg of DNA in Tris buffer $585
Add to Cart

Backbone

  • Vector backbone
    pcDNA3
  • Backbone manufacturer
    Invitrogen
  • Backbone size w/o insert (bp) 5350
  • Vector type
    Mammalian Expression, Luciferase
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    RLUC - IRES - FLUC
  • Alt name
    Renilla luciferase - IRES - Firefly luciferase
  • Insert Size (bp)
    3370
  • GenBank ID
    RLUC (1.13.12.5), FLUC (1.13.12.7), PVIRES (M12197)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site NheI (not destroyed)
  • 3′ cloning site XhoI (not destroyed)
  • 5′ sequencing primer CMV-F
  • 3′ sequencing primer BGH-rev
  • (Common Sequencing Primers)

Terms and Licenses

Trademarks:

  • Zeocin® is an InvivoGen trademark.

Depositor Comments

The bicistronic reporter plasmid pcDNA3-RLUC-POLIRES-FLUC was generated by subcloning the coding sequence for firefly luciferase (FLUC) from pGEM-LUC (Promega Biotech Inc.), the coding sequence for Renilla reniformis luciferase (RLUC) from pRL-CMV (Promega) and the complete poliovirus type II Lansing 5′-untranslated region (UTR) (poliovirus internal ribosome entry site, POLIRES, nucleotides 1–737) into pcDNA3.

Translation of the RLUC cistron is cap-dependent, whereas that of the FLUC cistron is directed by the poliovirus IRES and is therefore cap-independent.

RLUC is between NheI and XbaI. Polio IRES is between HindIII and EcoRV. FLUC is between BamHI and XhoI. See Author's Map for more detail.

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

  • Amount 50 μg
  • Concentration 1 μg/μL
  • Pricing $495 USD
  • Storage 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

  • Amount 100 μg
  • Concentration 1 μg/μL
  • Pricing $585 USD
  • Storage 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pcDNA3 RLUC POLIRES FLUC was a gift from Nahum Sonenberg (Addgene plasmid # 45642 ; http://n2t.net/addgene:45642 ; RRID:Addgene_45642)

  • For your References section:

    4E-BP3, a new member of the eukaryotic initiation factor 4E-binding protein family. Poulin F, Gingras AC, Olsen H, Chevalier S, Sonenberg N. J Biol Chem. 1998 May 29;273(22):14002-7. 10.1074/jbc.273.22.14002 PubMed 9593750
Related items:
Commonly requested with: