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pcDNA-NES-ZapCmR2
(Plasmid #59015)

Full plasmid sequence is not available for this item.

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 59015 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pcDNA3.1(+)
  • Backbone manufacturer
    Invitrogen
  • Backbone size w/o insert (bp) 5500
  • Total vector size (bp) 7200
  • Modifications to backbone
    Leu-rich NES sequence added upstream of BamHI site
  • 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
    NES-ZapCmR2 genetically encoded cytosolic Zn(II) sensor
  • Species
    Synthetic
  • Insert Size (bp)
    1689
  • Mutation
    See comment
  • Promoter CMV
  • Tag / Fusion Protein
    • Nuclear Exclusion Signal (N terminal on backbone)

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, fluorescent biosensor. Contains nuclear exclusion signal sequence at N-terminus. Zn(II)-binding domain derived from the first 2 zinc fingers of Zap 1 from Saccharomyces cerevisiae. Contains Clover and mRuby fluorescent proteins.

The Zn(II) binding domain of ZapCmR2 is derived from ZapCmR1.1. The first cysteine in the second zinc finger is mutated to histidine.

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-NES-ZapCmR2 was a gift from Amy Palmer (Addgene plasmid # 59015 ; http://n2t.net/addgene:59015 ; RRID:Addgene_59015)
  • For your References section:

    New alternately colored FRET sensors for simultaneous monitoring of Zn(2)(+) in multiple cellular locations. Miranda JG, Weaver AL, Qin Y, Park JG, Stoddard CI, Lin MZ, Palmer AE. PLoS One. 2012;7(11):e49371. doi: 10.1371/journal.pone.0049371. Epub 2012 Nov 16. 10.1371/journal.pone.0049371 PubMed 23173058