PurposeCre-dependently expresses the fusion protein EGFPL10a
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||98747||Standard format: Plasmid sent in bacteria as agar stab||1||$85|
|AAV5||98747-AAV5||Virus (100 µL at titer ≥ 7×10¹² vg/mL) and Plasmid.||$405|
This material is available to academics and nonprofits only.
Vector backbonepAAV-EF1a-double floxed-hChR2(H134R)-mCherry-WPRE-HGHpA
Backbone manufacturerKarl Deisseroth
- Backbone size w/o insert (bp) 5601
- Total vector size (bp) 7008
Modifications to backboneN/A
Vector typeMammalian Expression, AAV, Cre/Lox
Growth in Bacteria
Bacterial Resistance(s)Ampicillin, 100 μg/mL
Growth Strain(s)NEB Stable
Copy numberHigh Copy
SpeciesM. musculus (mouse); A. victoria (jellyfish)
Insert Size (bp)1407
- Promoter EF1a
/ Fusion Protein
- GFP (N terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site Asci (not destroyed)
- 3′ cloning site NheI (not destroyed)
- 5′ sequencing primer EF1a-Fwd
- 3′ sequencing primer WPRE-Rev (Common Sequencing Primers)
A portion of this plasmid was derived from a plasmid made byEric Schmidt, Nathaniel Heintz
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
Information for AAV5 (Catalog # 98747-AAV5) ( Back to top )
Ready-to-use AAV5 particles produced from pAAV-FLEX-EGFPL10a (#98747). In addition to the viral particles, you will also receive purified pAAV-FLEX-EGFPL10a plasmid DNA.Cre-dependent EGFPL10a (EGFP-tagged ribosomal protein) expression for translating ribosome affinity purification (TRAP). These AAV preparations are suitable purity for injection into animals.
- Volume 100 µL
- Titer ≥ 7×10¹² vg/mL
- Pricing $375 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 EGFP (Cre-dependent)
- 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 0.1-0.8% of viral particles in our typical production protocol. This can lead to a small number of cells exhibiting Cre-independent transgene expression in vivo. To address this, it is necessary to optimize the injection volume and viral titer to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral particle 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-FLEX-EGFPL10a was a gift from Nathaniel Heintz & Alexander Nectow & Eric Schmidt (Addgene plasmid # 98747 ; http://n2t.net/addgene:98747 ; RRID:Addgene_98747)
For viral preps, please replace (Addgene plasmid # 98747) in the above sentence with: (Addgene viral prep # 98747-AAV5)
For your References section:Rapid Molecular Profiling of Defined Cell Types Using Viral TRAP. Nectow AR, Moya MV, Ekstrand MI, Mousa A, McGuire KL, Sferrazza CE, Field BC, Rabinowitz GS, Sawicka K, Liang Y, Friedman JM, Heintz N, Schmidt EF. Cell Rep. 2017 Apr 18;19(3):655-667. doi: 10.1016/j.celrep.2017.03.048. 10.1016/j.celrep.2017.03.048 PubMed 28423326