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PurposeEncodes a BRET-based activity sensor for the heterotrimeric G protein G15. Composed of the subunits G alpha 15 (GNA15) tagged with NanoLuciferase, G beta 3 (GNB3) and Venus-tagged G gamma 9 (GNG9).
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 168126 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backboneClontech-style N1
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Backbone manufacturerClontech
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Vector typeMammalian Expression, Luciferase
Growth in Bacteria
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Bacterial Resistance(s)Kanamycin, 50 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 nameGNA15, GNB3, GNG9
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SpeciesH. sapiens (human)
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MutationNLuc is inserted at E244/N245 within GNA15
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Entrez GeneGNA15 (a.k.a. GNA16, HG1L)
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Entrez GeneGNB3 (a.k.a. CSNB1H, HG2D)
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Entrez GeneGNGT2 (a.k.a. G-GAMMA-8, G-GAMMA-C, GNG9, GNGT8, HG3I)
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Tag
/ Fusion Protein
- cpVenus on GNG9 (N terminal on insert)
Cloning Information
- Cloning method Unknown
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.
Depositor Comments
Please visit https://www.biorxiv.org/content/10.1101/2021.02.05.429900v1 for bioRxiv preprint.
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:
G15-CASE was a gift from Gunnar Schulte (Addgene plasmid # 168126 ; http://n2t.net/addgene:168126 ; RRID:Addgene_168126) -
For your References section:
Quantitative assessment of constitutive G protein-coupled receptor activity with BRET-based G protein biosensors. Schihada H, Shekhani R, Schulte G. Sci Signal. 2021 Sep 7;14(699):eabf1653. doi: 10.1126/scisignal.abf1653. Epub 2021 Sep 7. 10.1126/scisignal.abf1653 PubMed 34516756