pcDNA3.1(+)-RasAR-NES
(Plasmid #205567)

• Purpose: Genetically encoded FRET-based biosensor for monitoring cellular Ras GTPase activity. Targeted to cytosol.
• Depositing Lab: Jin Zhang
• Publication: Weeks et al J Am Chem Soc. 2022 Sep 28;144(38):17432-17440. doi: 10.1021/jacs.2c05203. Epub 2022 Sep 19.

Backbone

• Vector backbone: pcDNA3.1(+)
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5363
• Total vector size (bp): 7151
• 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: RasAR-NES
• Alt name: Cerulean3-RBD(Raf1 51-131)-pseudoligand-YPet-NES
• Species: H. sapiens (human), Synthetic
• Insert Size (bp): 1788
• Entrez Gene: RAF1 (a.k.a. CMD1NN, CRAF, NS5, Raf-1, c-Raf)
• Promoter: CMV
• Tag / Fusion Protein:
  • Nuclear export signal (NES) (C terminal on insert)

Cloning Information

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

Resource Information

• Supplemental Documents:
  • pcDNA3.1(+)-RasAR-NES.gbk

Terms and Licenses

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