MSCV-Luciferase-IRES-NrasG12D
(Plasmid #60834)

• Purpose: Mouse NrasG12D was amplified and mutated by PCR from IMAGE clone 6475312 (Invitrogen) and cloned downstream from the IRES; subsequently Luciferase was cloned upstream of the IRES
• Depositing Lab: Scott Lowe
• Publication: Zuber et al Genes Dev. 2009 Apr 1;23(7):877-89. doi: 10.1101/gad.1771409.

Backbone

• Vector backbone: MSCV
• Backbone manufacturer: Clonetech
• Modifications to backbone: All retroviruses were constructed in the MSCV backbone (Clontech). MSCV-IRES-Luciferase was generated by replacing GFP in MSCV-IRES-GFP with an NcoI/SalI fragment from pGL3-Basic (Promega) after excision of the 3′polyA signal. Mouse NrasG12D was amplified and mutated by PCR from IMAGE clone 6475312 (Invitrogen) and cloned downstream from the IRES; subsequently Luciferase was cloned upstream of the IRES.
• Vector type: Retroviral

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Top10
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: NrasG12D
• gRNA/shRNA sequence: NrasG12D
• Species: M. musculus (mouse)
• Entrez Gene: Nras (a.k.a. N-ras)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Eco (unknown if destroyed)
• 3′ cloning site: Xho (unknown if destroyed)
• 5′ sequencing primer: MSCV

Resource Information

• Supplemental Documents:
  • Gene Bank

Terms and Licenses

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