PKAsub(TA)-TagRFPT-CAAX
(Plasmid #87740)

• Purpose: Negative-control PKA substrate fragment for bimFLINC-AKAR, targeted to plasma membrane; use in conjunction with Dronpa-FHA1 (Addgene #87711).
• Depositing Lab: Jin Zhang
• Publication: Mo et al Nat Methods. 2017 Apr;14(4):427-434. doi: 10.1038/nmeth.4221. Epub 2017 Mar 13.

Backbone

• Vector backbone: pcDNA3
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5500
• Total vector size (bp): 6255
• 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: PKAsub(TA)-TagRFPT-CAAX
• Species: Synthetic
• Insert Size (bp): 784
• Mutation: Target Thr residue in substrate domain mutated to Ala.
• Promoter: CMV
• Tags / Fusion Proteins:
  • C-terminal targeting sequence from Kras (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-PKAsub(TA)-TagRFPT-CAAX.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Not Available to Industry