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ISceI-GR-RFP
(Plasmid #17654)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 17654 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pDsRed-Monomer-C1
  • Backbone manufacturer
    Clontech
  • Backbone size w/o insert (bp) 4700
  • Vector type
    Mammalian Expression

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    I-SceI-GRLBD
  • Alt name
    SCEI
  • Alt name
    GR ligand binding domain
  • Alt name
    glucocorticoid receptor
  • Species
    R. norvegicus (rat), S. cerevisiae (budding yeast)
  • Mutation
    C655G mutation in the glucocorticoid receptor (see comments)
  • Entrez Gene
    SCEI (a.k.a. Q0160)
  • Entrez Gene
    Nr3c1 (a.k.a. GR, Gcr, Grl)
  • Tag / Fusion Protein
    • mRFP (N terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site SacI (not destroyed)
  • 3′ cloning site BamHI (not destroyed)
  • 5′ sequencing primer DsRed1-C
  • 3′ sequencing primer SV40pA-R
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

The Ligand Binding
Domain (LBD) of GR was amplified from pCI-nGFP-C656G GR (gift from G. Hager) and cloned into pEGFP-C2
(Clontech) in the SacII and BamHI sites. The C656G mutation of GR that makes it more responsive to ligand and
trasnfers it more quickly to the nucleus. Similarly, the I-SceI cDNA was amplified from HA-I-SceI and cloned into the previously described
GFP-GRLBD into the SacI and SalI restriction sites. Finally, the chimeric I-SceI–GR cDNA was subcloned into pDsRed-Monomer-C1 (Clontech) into the SacI and BamHI sites.

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:

    ISceI-GR-RFP was a gift from Tom Misteli (Addgene plasmid # 17654 ; http://n2t.net/addgene:17654 ; RRID:Addgene_17654)
  • For your References section:

    Positional stability of single double-strand breaks in mammalian cells. Soutoglou E, Dorn JF, Sengupta K, Jasin M, Nussenzweig A, Ried T, Danuser G, Misteli T. Nat Cell Biol. 2007 Jun . 9(6):675-82. 10.1038/ncb1591 PubMed 17486118