|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||17654||Standard format: Plasmid sent in bacteria as agar stab||1||$85|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 4700
Vector typeMammalian Expression
Growth in Bacteria
Bacterial Resistance(s)Kanamycin, 50 μg/mL
Copy numberHigh Copy
Alt nameGR ligand binding domain
Alt nameglucocorticoid receptor
SpeciesR. norvegicus (rat), S. cerevisiae (budding yeast)
MutationC655G mutation in the glucocorticoid receptor (see comments)
Entrez GeneSCEI (a.k.a. Q0160)
Entrez GeneNr3c1 (a.k.a. GR, Gcr, Grl)
/ Fusion Protein
- mRFP (N terminal on backbone)
- 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)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
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.
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