This website uses cookies to ensure you get the best experience. By continuing to use this site, you agree to the use of cookies.

Please note: Your browser does not support the features used on Addgene's website. You may not be able to create an account or request plasmids through this website until you upgrade your browser. Learn more

Please note: Your browser does not fully support some of the features used on Addgene's website. If you run into any problems registering, depositing, or ordering please contact us at [email protected] Learn more

pcDNA3 RLUC POLIRES FLUC
(Plasmid #45642)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 45642 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

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
  • 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)

Resource Information

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.

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