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PurposePlasma membrane-targeted FRET biosensor for monitoring mTORC1 kinase activity.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 64930 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepcDNA3
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Backbone manufacturerInvitrogen
- Backbone size w/o insert (bp) 5446
- Total vector size (bp) 7257
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Vector typeMammalian Expression
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Selectable markersNeomycin (select with G418)
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameLyn-TORCAR
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SpeciesH. sapiens (human), Synthetic
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Insert Size (bp)1842
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GenBank ID
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Entrez GeneEIF4EBP1 (a.k.a. 4E-BP1, 4EBP1, BP-1, PHAS-I)
- Promoter CMV
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Tags
/ Fusion Proteins
- Cerulean (N terminal on insert)
- YPet (C terminal on insert)
- N-terminal targeting sequence from Lyn kinasen kinase (N terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site HindIII (not destroyed)
- 3′ cloning site EcoRI (not destroyed)
- 5′ sequencing primer T7 forward
- 3′ sequencing primer BGH reverse (Common Sequencing Primers)
Resource Information
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Supplemental Documents
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
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.
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For your Materials & Methods section:
pcDNA3-PM-TORCAR was a gift from Jin Zhang (Addgene plasmid # 64930 ; http://n2t.net/addgene:64930 ; RRID:Addgene_64930) -
For your References section:
Dynamic Visualization of mTORC1 Activity in Living Cells. Zhou X, Clister TL, Lowry PR, Seldin MM, Wong GW, Zhang J. Cell Rep. 2015 Mar 11. pii: S2211-1247(15)00177-1. doi: 10.1016/j.celrep.2015.02.031. 10.1016/j.celrep.2015.02.031 PubMed 25772363