PurposeExpresses dark mutant PA-DAD in mammalian cells, an optogenetic tool to activate endogenous diaphanous related formins in live cells using light with LOV2 in the closed conformation
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||53096||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 5238
Vector typeMammalian Expression, Bacterial Expression, Insect Expression
Growth in Bacteria
Bacterial Resistance(s)Ampicillin, 100 μg/mL
Copy numberHigh Copy
Alt namemVenus - 3x GS linker - LOV2-Jα (404-546) - 3x GS linker - DAD (1177-1222)
Alt nameLOV2 "dark state” (closed conformation) mutant
SpeciesM. musculus (mouse); Avena sativa
MutationL47F and C49S in Venus; C450M mutation in LOV2
/ Fusion Proteins
- 6x His (N terminal on backbone)
- mVenus (N terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (not destroyed)
- 3′ cloning site HindIII (not destroyed)
- 5′ sequencing primer pTriExUP
- 3′ sequencing primer pTriExDOWN (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
Expresses photoactivatable (PA) Diaphanous auto-regulatory domain (DAD) consisting of a fusion of mVenus, dark LOV2 Jα protein domain of Avena sativa Phototropin1, and Diaphanous auto-regulatory domain of mouse Diaphanous-related formin-1 (mDia1)
Please note that the C450M mutation in LOV2 creates a "dark state" mutant. See Yi et al., ACS Synth Biol. 2014 Nov 21;3(11):788-95 for more information.
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:PA-DAD-dark was a gift from Klaus Hahn & Ronen Zaidel-Bar (Addgene plasmid # 53096 ; http://n2t.net/addgene:53096 ; RRID:Addgene_53096)
For your References section:An optogenetic tool for the activation of endogenous diaphanous-related formins induces thickening of stress fibers without an increase in contractility. Rao MV, Chu PH, Hahn KM, Zaidel-Bar R. Cytoskeleton (Hoboken). 2013 Jul;70(7):394-407. doi: 10.1002/cm.21115. Epub 2013 May 24. 10.1002/cm.21115 PubMed 23677607