pcDNA-D1ER
(Plasmid #36325)

• Depositing Labs: Amy Palmer, Roger Tsien
• Publication: Palmer et al Proc Natl Acad Sci U S A. 2004 Dec 14;101(50):17404-9. Epub 2004 Dec 7.

Backbone

• Vector backbone: pcDNA3
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5400
• Total vector size (bp): 7400
• 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): DH10B
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: D1ER second generation cameleon (calcium sensor) targeted to ER
• Species: Synthetic
• Insert Size (bp): 1983
• GenBank ID: AY796115.1
• Promoter: CMV
• Tag / Fusion Protein:
  • KDEL (C terminal on insert)

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

• Articles Citing this Plasmid:
  • 26 References

Terms and Licenses

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

Depositor Comments

Genetically encoded, ratiometric, calcium biosensor that contains enhanced CFP and Citrine. D1ER does not bind wild-type calmodulin. It has a Kd1' of 0.8 uM and a Kd2' of 60 uM.

D1ER is retained in the ER with a C-terminal KDEL sequence.

See also Palmer & Tsien (2006) Nature Protocols.