pcDNA3 RLUC POLIRES FLUC
PurposeBicistronic reporter plasmid expressing Renilla luciferase and firefly luciferase in mammalian cells.
|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 size w/o insert (bp) 5350
Vector typeMammalian Expression, Luciferase
Selectable markersNeomycin (select with G418)
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert nameRLUC - IRES - FLUC
Alt nameRenilla luciferase - IRES - Firefly luciferase
Insert Size (bp)3370
GenBank IDRLUC (18.104.22.168), FLUC (22.214.171.124), PVIRES (M12197)
- 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)
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.
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