pcDNA3-PDE4D2cat-ICUE4
(Plasmid #181847)

• Purpose: ICUE4 cAMP sensor tethered to the catalytic domain of PDE4D2.
• Depositing Lab: Jin Zhang
• Publication: Zhang et al Cell. 2020 Sep 17;182(6):1531-1544.e15. doi: 10.1016/j.cell.2020.07.043. Epub 2020 Aug 25.

Backbone

• Vector backbone: pcDNA3
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5403
• Total vector size (bp): 10110
• 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: PDE4D2cat-ICUE4
• Alt name: PDE4D2(86-418)-ECFP-Epac1(149-881; Q270E)-cpVenus(L194)
• Species: H. sapiens (human)
• Insert Size (bp): 4707
• Mutation: Glutamine 122 in Epac1 domain mutated to glutamate (corresponds to Q270E in full-length Epac).
• Entrez Gene: RAPGEF3 (a.k.a. CAMP-GEFI, EPAC, EPAC1, HSU79275, bcm910)
• Entrez Gene: PDE4D (a.k.a. ACRDYS2, DPDE3, HSPDE4D, PDE43, PDE4DN2, STRK1)
• Promoter: CMV
• Tags / Fusion Proteins:
  • ECFP (N terminal on insert)
  • cpVenus(L194) (C terminal on insert)
  • PDE4D2(86-418) (N terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CMV forward
• 3′ sequencing primer: BGH reverse

Resource Information

• Supplemental Documents:
  • pcDNA3-PDE4D2cat-ICUE4.gb
• A portion of this plasmid was derived from a plasmid made by: PDE4D2 cDNA was provided by Hengming Ke (UNC Chapel Hill).

Terms and Licenses

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