pcDNA3 RLUC POLIRES FLUC
(Plasmid #45642)

• Purpose: Bicistronic reporter plasmid expressing Renilla luciferase and firefly luciferase in mammalian cells.
• Depositing Lab: Nahum Sonenberg
• Publication: Poulin et al J Biol Chem. 1998 May 29;273(22):14002-7.

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

Resource Information

• Articles Citing this Plasmid:
  • 19 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Luciferase Limited Use Label License
  • Renilla Luciferase Limited Use Label License
• Industry Terms:
  • Not Available to Industry

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.