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pN1-Lck-GCaMP3
(Plasmid #26974)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 26974 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pEGFP-N1
  • Backbone manufacturer
    Clontech
  • Backbone size w/o insert (bp) 3950
  • Vector type
    Mammalian Expression
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    Lck-GCaMP3
  • Alt name
    GCaMP
  • Alt name
    GCaMP3
  • Alt name
    Lck-GCaMP3
  • Insert Size (bp)
    1480
  • Mutation
    Plasmid contains N-terminal 26 amino acid membrane targeting sequence of Lck (Src tyrosine kinase) fused to GCaMP3. 6xHis and Xpress (FLAG-like) epitope tags contained within 'RSET' sequence, making up the linker region between Lck and GCaMP3
  • Tags / Fusion Proteins
    • His (N terminal on backbone)
    • Xpress (N terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site BglII (not destroyed)
  • 3′ cloning site NotI (not destroyed)
  • 5′ sequencing primer ACGGTGGGAGGTCTATATAAGCAG AG
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

Sequence is identical to pN1-Lck-GCaMP2 http://www.addgene.org/24794 , except for three mutations in GCaMP2 to make GCaMP3: M153K (EGFP); T203V (EGFP) and N60D (CaM).

Note that the RSET tag refers to the first 37 amino acids of the insert, which include the His tag, T7 tag and Xpress tag: MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVD

A genetically targeted optical sensor to monitor calcium signals in astrocyte processes. Shigetomi E., et al. Nat. Neurosci. 2010 Jun;13(6):759-66.

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:

    pN1-Lck-GCaMP3 was a gift from Baljit Khakh (Addgene plasmid # 26974 ; http://n2t.net/addgene:26974 ; RRID:Addgene_26974)
  • For your References section:

    Monitoring astrocyte calcium microdomains with improved membrane targeted GCaMP reporters. Shigetomi E, Kracun S, Khakh BS.. Neuron Glia Biol. 2010 Dec 16:1-9. [Epub ahead of print] 10.1017/S1740925X10000219 PubMed 21205365