Purposegenetically-encoded fluorescent voltage indicator
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||85843||Standard format: Plasmid sent in bacteria as agar stab||1||$85|
This material is available to academics and nonprofits only.
Vector backbonepCS2+ hSyn1
- Backbone size w/o insert (bp) 4372
- Total vector size (bp) 5833
Vector typeMammalian Expression
Growth in Bacteria
Bacterial Resistance(s)Ampicillin, 100 μg/mL
Copy numberHigh Copy
Gene/Insert nameGEVI Marina
Insert Size (bp)1461
MutationCi-VSP contains R217Q mutation; super ecliptic pHluorin contains D147A, H148A, Y200V, A227D mutations. These mutations cause depolarization dependent increase in fluorescence intensity. The A227D mutation increases the fluorescence response magnitude. We replaced residues 71-91 within CiVSD with an amino acid sequence (KSRITSEGEYIPLDQIDINV) from the Kir2.1 channel Golgi-to-plasma membrane trafficking signal. The Kir 2.1 channel ER export signal sequence (FCYENEV) was added to C-terminus of super ecliptic pHluorin. The sequence is codon optimized using mammalian codon preference.
GenBank IDAB183035 AY533296
- Promoter hSyn1
/ Fusion Protein
- mCherry (C terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site KpnI (not destroyed)
- 3′ cloning site SphI (not destroyed)
- 5′ sequencing primer AGTCGTGTCGTGCCTGAGAG
- 3′ sequencing primer ccttgagcatctgacttctggcta (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
A portion of this plasmid was derived from a plasmid made by Dr. Atsushi Miyawaki, Laboratory for Cell Function Dynamics, Brain Science Institute, RIKEN, 2-1 Hirosawa, Saitama 351-0198, Japan.
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:hSyn Marina-T2A-nls-mCherry was a gift from Vincent Pieribone (Addgene plasmid # 85843 ; http://n2t.net/addgene:85843 ; RRID:Addgene_85843)
For your References section:Directed evolution of key residues in fluorescent protein inverses the polarity of voltage sensitivity in the genetically-encoded indicator ArcLight. Platisa J, Vasan G, Yang A, Pieribone VA. ACS Chem Neurosci. 2017 Jan 3. doi: 10.1021/acschemneuro.6b00234. 10.1021/acschemneuro.6b00234 PubMed 28045247