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pcDNA-D3cpv
(Plasmid #36323)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 36323 Standard format: Plasmid sent in bacteria as agar stab 1 $94
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DNA 36323-D50 50 μg of DNA in Tris buffer $495
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DNA 36323-D100 100 μg of DNA in Tris buffer $585
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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)
    1975
  • GenBank ID
    DQ479426.1
  • 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
  • (Common Sequencing Primers)

Terms and Licenses

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

Trademarks:

  • Zeocin® is an InvivoGen trademark.

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.

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

  • Amount 50 μg
  • Concentration 1 μg/μL
  • Pricing $495 USD
  • Storage 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

  • Amount 100 μg
  • Concentration 1 μg/μL
  • Pricing $585 USD
  • Storage 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pcDNA-D3cpv was a gift from Amy Palmer & Roger Tsien (Addgene plasmid # 36323 ; http://n2t.net/addgene:36323 ; RRID:Addgene_36323)

  • For your References section:

    Ca2+ indicators based on computationally redesigned calmodulin-peptide pairs. Palmer AE, Giacomello M, Kortemme T, Hires SA, Lev-Ram V, Baker D, Tsien RY. Chem Biol. 2006 May;13(5):521-30. 10.1016/j.chembiol.2006.03.007 PubMed 16720273
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