GPKAnes
(Plasmid #61092)

• Purpose: GFP-tagged PKA catalytic subunit targeted to the extranuclear cytoplasm by addition of a nuclear export signal
• Depositing Lab: Steven Green
• Publication: Bok et al J Neurosci. 2003 Feb 1;23(3):777-87.

Backbone

• Vector backbone: EGFP-C1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 4731
• Modifications to backbone: PCR-amplified from RSV-PKA-C-2 (Howard et al. 1991. JBC 266:10189), adding 5’-BspE1 and 3’-XhoI sites, and ligated into EGFP-C1. To make GPKAnes, a double-stranded oligonucleotide encoding the MEK (mitogen-activated protein kinase kinase) nes (QALQKKLEELELDE) (Wu et al., 1992) was added to the C-terminus of the GPKA between HindIII and BamHI.
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH10B
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: pkac2
• Species: Cricetulus griseus
• Promoter: CMV
• Tag / Fusion Protein:
  • EGFP (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BspEI (unknown if destroyed)
• 3′ cloning site: BamHI (unknown if destroyed)
• 5′ sequencing primer: EGFP-C
• 3′ sequencing primer: SV40 poly A Reverse

Resource Information

• A portion of this plasmid was derived from a plasmid made by: See publication for cloning details.

Terms and Licenses

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

Depositor Comments

The Y307H found in Addgene's QC sequence has no known functional consequence.