pcDNA3-Lyn-dPlcR
(Plasmid #181832)

• Purpose: Genetically encoded single-color sensor for monitoring plasma membrane PI(4,5)P2 dynamics in living cells. Based on dimerization-dependent green fluorescent protein (ddGFP).
• Depositing Lab: Jin Zhang
• Publication: Hertel et al ACS Chem Biol. 2020 Jan 17;15(1):33-38. doi: 10.1021/acschembio.9b00691. Epub 2020 Jan 6.

Backbone

• Vector backbone: pcDNA3
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5415
• Total vector size (bp): 7389
• 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: Lyn-PlcR
• Alt name: Lyn-GFP_A-PH(PLCdelta)-pseudoligand-FP_B
• Insert Size (bp): 1974
• Promoter: CMV
• Tags / Fusion Proteins:
  • ddGFP-A (N terminal on insert)
  • ddFP-B (C terminal on insert)
  • N-terminal targeting sequence from Lyn kinase (N terminal on insert)

Cloning Information

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

Resource Information

• Supplemental Documents:
  • pcDNA3-Lyn-dPlcR.gb

Terms and Licenses

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