-
PurposeA FRET sensor for Galphai3 activation, encoded on a single plasmid.
-
Depositing Lab
-
Depositing OrganizationUniversity of Amsterdam
Located in Netherlands -
Citations
-
Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 69625 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 69625-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 69625-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
-
Vector backboneClontech-style N1
-
Backbone manufacturerClontech
- Backbone size w/o insert (bp) 4000
- Total vector size (bp) 8445
-
Vector typeMammalian Expression
-
Selectable markersNeomycin (select with G418)
Growth in Bacteria
-
Bacterial Resistance(s)Kanamycin, 50 μg/mL
-
Growth Temperature37°C
-
Growth Strain(s)DH5alpha
-
Copy numberHigh Copy
Gene/Insert 1
-
Gene/Insert nameGbeta-2A-cpV-Ggamma2
-
SpeciesSynthetic
-
Insert Size (bp)2046
- Promoter CMV
-
Tag
/ Fusion Protein
- ciruclar permutated Venus (cp173V)
Cloning Information for Gene/Insert 1
- Cloning method Unknown
- 5′ sequencing primer CMV Forward
- 3′ sequencing primer IRES-R
- (Common Sequencing Primers)
Gene/Insert 2
-
Gene/Insert nameGalphai3-mTurquoise2
-
SpeciesSynthetic
-
Insert Size (bp)1770
- Promoter CMV (IRES)
-
Tag
/ Fusion Protein
- mTurquoise2
Cloning Information for Gene/Insert 2
- Cloning method Unknown
- 5′ sequencing primer IRES-F
- (Common Sequencing Primers)
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
DNA (Catalog # 69625-D50)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 50 μg
- Concentration 1 μg/μL
- Pricing $495 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
DNA (Catalog # 69625-D100)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 100 μg
- Concentration 1 μg/μL
- Pricing $585 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
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:
Gβ-2A-cpV-Gγ2-IRES-Gαi3-mTq2 was a gift from Dorus Gadella (Addgene plasmid # 69625 ; http://n2t.net/addgene:69625 ; RRID:Addgene_69625) -
For your References section:
A New Generation of FRET Sensors for Robust Measurement of Galphai1, Galphai2 and Galphai3 Activation Kinetics in Single Cells. van Unen J, Stumpf AD, Schmid B, Reinhard NR, Hordijk PL, Hoffmann C, Gadella TW Jr, Goedhart J. PLoS One. 2016 Jan 22;11(1):e0146789. doi: 10.1371/journal.pone.0146789. eCollection 2016. PONE-D-15-42389 [pii] PubMed 26799488
