PurposeAAV-mediated expression of Jaws-KGC-tdTomato-ER2 under the CAG promoter, in floxed/reversed (Cre-dependent) manner.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||84446||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Vector backboneAAV with CAG promter
- Backbone size w/o insert (bp) 4897
- Total vector size (bp) 7267
Vector typeMammalian Expression, AAV
Growth in Bacteria
Growth Strain(s)NEB Stable
Growth instructionsCan use Stbl3 at 30°C (use carbenicillin if using Stbl3) or DH5a at 37°C
Copy numberLow Copy
SpeciesHaloarcula salinarum (stain Shark)
Insert Size (bp)2370
- Promoter CAG
/ Fusion Proteins
- KGC (C terminal on insert)
- tdTomato (C terminal on insert)
- ER2 (C terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site NheI (not destroyed)
- 3′ cloning site AscI (not destroyed)
- 5′ sequencing primer tcggcttctggcgtgtgac
- 3′ sequencing primer aaagagacagcaaccagg (Common Sequencing Primers)
The plasmid is fully sequenced in the coding sequence regions (opsin-fluorophore and important flanking regions). Multiple digestions were done to verify the vector structure. The construct and the virus were both tested in vitro.
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-CAG-FLEX-rc [Jaws-KGC-tdTomato-ER2] was a gift from Edward Boyden (Addgene plasmid # 84446 ; http://n2t.net/addgene:84446 ; RRID:Addgene_84446)
For your References section:Noninvasive optical inhibition with a red-shifted microbial rhodopsin. Chuong AS, Miri ML, Busskamp V, Matthews GA, Acker LC, Sorensen AT, Young A, Klapoetke NC, Henninger MA, Kodandaramaiah SB, Ogawa M, Ramanlal SB, Bandler RC, Allen BD, Forest CR, Chow BY, Han X, Lin Y, Tye KM, Roska B, Cardin JA, Boyden ES. Nat Neurosci. 2014 Aug;17(8):1123-9. doi: 10.1038/nn.3752. Epub 2014 Jul 6. 10.1038/nn.3752 PubMed 24997763