pRK5-HA GST RagB 54L
Full plasmid sequence is not available for this item.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||19302||Standard format: Plasmid sent in bacteria as agar stab||1||$85|
This material is available to academics and nonprofits only.
Backbone manufacturerBD PharMingen
- Backbone size w/o insert (bp) 4754
Vector typeMammalian Expression
Growth in Bacteria
Bacterial Resistance(s)Ampicillin, 100 μg/mL
Copy numberHigh Copy
Gene/Insert nameRas-related GTP binding B
SpeciesH. sapiens (human)
Insert Size (bp)1040
Entrez GeneRRAGB (a.k.a. RAGB, bA465E19.1)
/ Fusion Proteins
- HA (N terminal on insert)
- GST (N terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site SalI (not destroyed)
- 3′ cloning site NotI (not destroyed)
- 5′ sequencing primer SP6, pGEX 5'
- 3′ sequencing primer pBABE 3' (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
Please note that this plasmid is actually quite toxic to cells. From the Sabatini lab:"it is a dominant negative and therefore inhibits the mTORC1 pathway and thus cells."
From the paper:
"To verify the actions of the Rags in a more physiological setting than that achieved by transient cDNA transfection, we generated HEK-293T cell lines stably expressing Rheb1, RagB, or RagBGTP (attempts to generate lines stably expressing RagBGDP failed)."
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:pRK5-HA GST RagB 54L was a gift from David Sabatini (Addgene plasmid # 19302 ; http://n2t.net/addgene:19302 ; RRID:Addgene_19302)
For your References section:The Rag GTPases bind raptor and mediate amino acid signaling to mTORC1. Sancak Y, Peterson TR, Shaul YD, Lindquist RA, Thoreen CC, Bar-Peled L, Sabatini DM. Science. 2008 Jun 13. 320(5882):1496-501. 10.1126/science.1157535 PubMed 18497260