PurposeIn the presence of Cre, the plasmid can be used to express GCaMP6s fused to the presynaptic protein synaptophysin. Drives enriched expression of GCaMP6s in neuronal axon boutons.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||105715||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
|AAV5||105715-AAV5||Virus (100 µL at titer ≥ 7×10¹² vg/mL) and Plasmid.|
This material is available to academics and nonprofits only.
Backbone manufacturerKarl Deisseroth
- Backbone size w/o insert (bp) 6109
- Total vector size (bp) 7465
Modifications to backboneThe post-transcriptional regulatory WPRE was replaced by WPRE3. The hGH polyA sequence was replaced by SV40pA.
Vector typeMammalian Expression, AAV, Cre/Lox
Growth in Bacteria
Growth Strain(s)NEB Stable
Copy numberHigh Copy
Alt namesynaptophysin-GCaMP3-K78H T302L R303P D380Y T381R S383T R392G
SpeciesR. norvegicus (rat); A. victoria (jellyfish)
Insert Size (bp)1274
- Promoter Ef1a, Human elongation factor-1 alpha
- Cloning method Restriction Enzyme
- 5′ cloning site AscI (not destroyed)
- 3′ cloning site RsrII (destroyed during cloning)
- 5′ sequencing primer acttctctccaaggtttgtc
- 3′ sequencing primer ttgatatcgaattcataact (Common Sequencing Primers)
A portion of this plasmid was derived from a plasmid made byGCaMP6s was synthesized de novo based on sequences published by Douglas Kim et al, Janelia Research Campus in Chen et al, 2013 PMID: 23868258. The Synaptophysin-GCaMP6s fusion sequence was previously published by Ofer Yizhar's lab in Mahn et al, 2016 PMID: 26950004
Terms and Licenses
- Not Available to Industry
Information for AAV5 (Catalog # 105715-AAV5) ( Back to top )
Ready-to-use AAV5 particles produced from pAAV-Ef1a-DIO-Synaptophysin-GCaMP6s (#105715). In addition to the viral particles, you will also receive purified pAAV-Ef1a-DIO-Synaptophysin-GCaMP6s plasmid DNA.EF1a-driven, Cre-dependent GCaMP6s fused to the presynaptic protein synaptophysin. These AAV preparations are suitable purity for injection into animals.
- Volume 100 µL
- Titer ≥ 7×10¹² vg/mL
- Pricing $350 USD for preparation of 100 µL virus + $30 USD for plasmid.
- Storage Store at -80℃. Thaw just before use and keep on ice.
- Shipment Viral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
Viral Production & Use
- Packaging Plasmids encode adenoviral helper sequences and AAV rep gene, AAV5 cap gene
- Buffer PBS + 0.001% Pluronic F-68 + 200 mM NaCl
- Serotype AAV5
- Purification Iodixanol gradient ultracentrifugation
Requestor is responsible for compliance with their institution's biosafety regulations. Lentivirus is generally considered BSL-2. AAV is generally considered BSL-1, but may require BSL-2 handling depending on the insert. Biosafety Guide
Terms and Licenses
- Not Available to Industry
Viral Quality Control
- Addgene ensures high quality viral vectors by optimizing and standardizing production protocols and performing rigorous quality control (QC) (see a list of our QC assays). The specific QC assays performed varies for each viral lot. To learn which specific QC assays were performed on your lot, please contact us.
- Titer: the exact titer of your sample will be reported on the tube. The titer you see listed on this page is the guaranteed minimum titer. See how titers are measured.
Visit our viral production page for more information.
Using FLEX vectors in vivo: LoxP sites in FLEX plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Cre-activated (i.e., "flipped") viral vectors. Addgene has measured this occurs in 0.01-0.03% of viral vectors in our typical production protocol. This can lead to a small number of cells exhibiting Cre-independent transgene expression in vivo. To address this, we recommend titrating to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral vector dosage in order to reduce the likelihood of Cre-independent expression.
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:pAAV-Ef1a-DIO-Synaptophysin-GCaMP6s was a gift from Rylan Larsen (Addgene plasmid # 105715 ; http://n2t.net/addgene:105715 ; RRID:Addgene_105715)
For viral preps, please replace (Addgene plasmid # 105715) in the above sentence with: (Addgene viral prep # 105715-AAV5)