pLenti-TO/CMV-puro-DD-PAmCherry1-KRas G12D
(Plasmid #177320)

• Purpose: Lentiviral vector expressing PAmCherry1-tagged KRas 4B G12D mutant, with an N-terminal dimerization domain (DD).
• Depositing Labs: Steven Chu, Xiaolin Nan
• Publication: Nan et al Proc Natl Acad Sci U S A. 2015 Jun 30;112(26):7996-8001.

Backbone

• Vector backbone: pLenti-TO/CMV-puro
• Backbone manufacturer: Addgene
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Growth instructions: One may also use the MDS42 or MDS69 strains from Scarab Genomics (https://www.scarabgenomics.com/products/clean-genome-e-coli/) when propagating these plasmids.
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: KRAS 4B
• Alt name: KRAS2
• Alt name: KRAS
• Species: H. sapiens (human)
• Insert Size (bp): 567
• Mutation: G12D
• Entrez Gene: KRAS (a.k.a. 'C-K-RAS, C-K-RAS, CFC2, K-RAS2A, K-RAS2B, K-RAS4A, K-RAS4B, K-Ras, K-Ras 2, KI-RAS, KRAS1, KRAS2, NS, NS3, OES, RALD, RASK2, c-Ki-ras, c-Ki-ras2)
• Promoter: TetOn-CMV
• Tags / Fusion Proteins:
  • PAmCherry1 (N terminal on insert)
  • FKBP-derived dimerization domain (DD, or DmrB) (N terminal on insert)

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: CMV_Forward

Resource Information

• Supplemental Documents:
  • pLenti-TOCMV-puro-DD-RKD.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The lentiviral backbone was created by Eric Campeau et al. (https://www.addgene.org/17293/) as a DESTination vector. The full insert was first created in an ENTRY clone (Invitrogen) and transferred to the DEST vector via gateway cloning.