pcDNA-lynD3cpV
(Plasmid #37472)

• Depositing Labs: Amy Palmer, Roger Tsien
• Publication: Palmer et al Chem Biol. 2006 May;13(5):521-30.

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): AAGCTTGCGGCCGCCAACATGGTGCGGGGTTCTCATCATCATCATCATCATGGTATGGCTAGCATGACTGGTGGACAGCAAATGGGTCGGGATCTGTACGACGATGACGATAAGGATCCC
• 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: D3cpv second generation cameleon (calcium sensor)
• Alt name: D family cameleon
• Species: Synthetic
• Insert Size (bp): 2049
• GenBank ID: DQ479428.1
• Promoter: CMV
• Tag / Fusion Protein:
  • the myristoylation and palmitoyla- tion sequence from lyn kinase (MGCIKSKRKDNLNDDGVDMKT) (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: 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 circularly permuted Venus 173. D3cpv does not bind wild-type calmodulin. It has a Kd' of 0.6 uM.

See also Palmer & Tsien (2006) Nature Protocols.