pcDNA-D1
(Plasmid #37480)

• 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): 7500
• Total vector size (bp): 9500
• Modifications to backbone: Note that the N-terminus of pcDNA3’ has the His tag of pRSETB inserted after the HindIII site (There is also a NotI site): AAGCTTGCGGCCGCCAACATGGTGC GGGGTTCTCATCATCATCATCATCAT GGTATGGCTAGCATGACTGGTGGACA GCAAATGGGTCGGGATCTGTACGACG ATGACGATAAGGATCCC
• 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: D1 second generation cameleon (calcium sensor)
• Alt name: D family cameleon
• Species: Synthetic
• Insert Size (bp): 1950
• GenBank ID:
• Promoter: CMV

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

• A portion of this plasmid was derived from a plasmid made by: Cloned by Amy Palmer in the lab of Roger Tsien.

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. D1 does not bind wild-type calmodulin. the apparent Kd1 (0.58), Kd2 (56.46).

See also Palmer & Tsien (2006) Nature Protocols.