pcDNA3.1(+)-ER-BSCaM2
(Plasmid #64855)

• Purpose: Endoplasmic reticulum (ER)-targeted version of a high-affinity FRET-based biosensor for monitoring Ca2+/CaM levels; targeted to cytosolic face of ER membrane.
• Depositing Lab: Jin Zhang
• Publication: Mehta et al eLife. 2014 Jul 23;3:e03765. doi: 10.7554/eLife.03765.

Backbone

• Vector backbone: pcDNA3.1+
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5428
• Total vector size (bp): 6967
• 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: CYP450-BSCaM2
• Species: Synthetic
• Insert Size (bp): 1611
• Promoter: CMV
• Tags / Fusion Proteins:
  • CFP (N terminal on insert)
  • YFP (N terminal on insert)
  • N-terminal 27 amino acids from CYP450 (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: T7 forward
• 3′ sequencing primer: BGH reverse

Resource Information

• Supplemental Documents:
  • pcDNA3.1(+)-ER-BSCaM2
• A portion of this plasmid was derived from a plasmid made by: The original BSCam2 construct was developed by Anthony Persechini’s lab (PMID: 12738782).

Terms and Licenses

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