pcDNA3-Lyso-mCherry-Rheb(C181S)
(Plasmid #192438)

• Purpose: Encodes mCherry-tagged, farnesylation-deficient Rheb C181S mutant targeted to lysosome membrane.
• Depositing Lab: Jin Zhang
• Publication: Zhong et al Cell Chem Biol. 2022 Jun 16;29(6):1037-1045.e4. doi: 10.1016/j.chembiol.2022.02.006. Epub 2022 Mar 15.

Backbone

• Vector backbone: pcDNA3
• Backbone size w/o insert (bp): 5415
• Total vector size (bp): 7962
• Vector type: Mammalian Expression
• 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: Lyso-mCherry-Rheb(C181S)
• Species: H. sapiens (human)
• Insert Size (bp): 2547
• Mutation: Rheb Cys 181 mutated to Ser; abolishes C-terminal farnesylation.
• Entrez Gene: RHEB (a.k.a. RHEB2)
• Promoter: CMV
• Tags / Fusion Proteins:
  • Full-length LAMP1 (N terminal on insert)
  • mCherry (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: CMV forward
• 3′ sequencing primer: BGH reverse

Resource Information

• Supplemental Documents:
  • pcDNA3-Lyso-mCherry-Rheb (C181S).gbk

Terms and Licenses

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