pcDNA3-GFP-GFP-FLARE-AKAR
(Plasmid #123332)

• Purpose: Green-fluorescent homoFRET/anisotropy-based biosensor for monitoring Protein Kinase A activity.
• Depositing Lab: Jin Zhang
• Publication: Ross et al eLife. 2018 Jul 3;7. pii: 35458. doi: 10.7554/eLife.35458

Backbone

• Vector backbone: pcDNA3
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5517
• Total vector size (bp): 7476
• 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: GFP-GFP-FLARE-AKAR
• Species: S. cerevisiae (budding yeast), Synthetic
• Insert Size (bp): 1959
• Promoter: CMV
• Tags / Fusion Proteins:
  • EGFP (N terminal on insert)
  • EGFP (C terminal on insert)
  • 6xHIS (N terminal on backbone)
  • T7 tag (gene 10 leader) (N terminal on backbone)
  • Xpress (TM) tag (N terminal on backbone)

Cloning Information

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

Resource Information

• Supplemental Documents:
  • pcDNA3-GFP-GFP-FLARE-AKAR.gb

Terms and Licenses

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