pcDNA3-AMPKsub-YPet-NES
(Plasmid #84635)

• Purpose: Encodes C-terminal (substrate) fragment of bimolecular AMPK/BRSK activity reporter (bimABKAR); cytosol targeted; use in conjunction with pcDNA3-Cerulean-FHA1-NES
• Depositing Lab: Jin Zhang
• Publication: Depry et al Chem Biol. 2015 Nov 19;22(11):1470-9. doi: 10.1016/j.chembiol.2015.10.004. Epub 2015 Nov 5.

Backbone

• Vector backbone: pcDNA3
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5500
• Total vector size (bp): 6297
• 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: AMPKsub-YPet-NES
• Alt name: bimABKAR-C
• Species: Synthetic
• Insert Size (bp): 807
• Promoter: CMV
• Tags / Fusion Proteins:
  • YPet (C terminal on insert)
  • Nuclear export signal (NES) (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: T7 forward
• 3′ sequencing primer: BGH reverse

Resource Information

• Supplemental Documents:
  • pcDNA3-AMPKsub-YPet-NES.gb

Terms and Licenses

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