Flag pLJM1 RagB 54L
(Plasmid #19314)

• Depositing Lab: David Sabatini
• Publication: Sancak et al Science. 2008 Jun 13. 320(5882):1496-501.

Backbone

• Vector backbone: pLJM1
• Backbone size w/o insert (bp): 7400
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Growth instructions: Grow in Exel10Gold or Stbl2 cells
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Ras-related GTP binding B
• Alt name: RRAGB
• Species: H. sapiens (human)
• Insert Size (bp): 1040
• Mutation: T54L
• Entrez Gene: RRAGB (a.k.a. RAGB, bA465E19.1)
• Tag / Fusion Protein:
  • Flag (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SalI (not destroyed)
• 3′ cloning site: EcoRV (not destroyed)
• 5′ sequencing primer: CMV-F

Resource Information

• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

pLJM1 is based on pLKO.1. pLJM1 was cut with AgeI and EcoRI and a Flag tag was cloned in. The cDNA was cloned into Flag-pLJM1 with SalI and EcoRV. The sequence across the MCS includes AgeI-Flag-SalI-gene-EcoRV-EcoRI

Note from the depositor: This plasmid does not express well. This is because it is a dominant negative and therefore inhibits the mTORC1 pathway and thus cells.