PurposeFlp recombinase dependent on GFP (Flp-DOG) for rAAV production and expression in mammalian cells
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||75469||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
|AAV1||75469-AAV1||Virus (100 µL at titer ≥ 1×10¹³ vg/mL) and Plasmid.|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 5329
- Total vector size (bp) 6704
Vector typeMammalian Expression, AAV, Synthetic Biology
Growth in Bacteria
Growth Strain(s)NEB Stable
Insert Size (bp)1986
- Promoter EF-1a
- Cloning method Unknown
- 5′ sequencing primer AGTGGGTGGAGACTGAAGTTAGG
- 3′ sequencing primer CACTGCACTCCAGCTTGGTTC (Common Sequencing Primers)
Information for AAV1 (Catalog # 75469-AAV1) ( Back to top )
Ready-to-use AAV1 particles produced from pAAV-EF1a-Flp-DOG-NW (#75469). In addition to the viral particles, you will also receive purified pAAV-EF1a-Flp-DOG-NW plasmid DNA.EF1a-driven expression of Flp recombinase dependent on GFP (Flp-DOG); to be coinjected with GFP (or for use with transgenic GFP animals). These AAV preparations are suitable purity for injection into animals.
- Volume 100 µL
- Titer ≥ 1×10¹³ 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, AAV1 cap gene
- Buffer PBS + 0.001% Pluronic F-68 + 200 mM NaCl
- Serotype AAV1
- 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
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.
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-Flp-DOG-NW was a gift from Connie Cepko (Addgene plasmid # 75469 ; http://n2t.net/addgene:75469 ; RRID:Addgene_75469)
For viral preps, please replace (Addgene plasmid # 75469) in the above sentence with: (Addgene viral prep # 75469-AAV1)
For your References section:Detection and manipulation of live antigen-expressing cells using conditionally stable nanobodies. Tang JC, Drokhlyansky E, Etemad B, Rudolph S, Guo B, Wang S, Ellis EG, Li JZ, Cepko CL. Elife. 2016 May 20;5. pii: e15312. doi: 10.7554/eLife.15312. 10.7554/eLife.15312 PubMed 27205882