Purpose(Empty Backbone) LINuS-flex: modular LINuS construct enabling introduction of any protein encoding sequence, NES and NLS. The sequence of interest replaces the LexA DBD-MBP-VP64 fragment.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||61349||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Vector typeMammalian Expression
- Promoter CMV
/ Fusion Proteins
- mCherry (C terminal on backbone)
- AsLOV2 wt (C terminal on backbone)
Growth in Bacteria
Copy numberHigh Copy
- Cloning method Restriction Enzyme
- 5′ sequencing primer CMV-F
- 3′ sequencing primer mCherry-R (Common Sequencing Primers)
LINuS-flex was designed to contain a spacer sequence flanked by BsmBI restriction sites between the VP64 transactivation domain and mCherry enabling scarless introduction of any NES sequence by standard oligo cloning.
A second spacer sequence flanked by BsaI restriction sites is located behind the truncated AsLOV2 domain and enables scarless introduction of any NLS sequence by standard oligo cloning.
Unique BamHI and SpeI sites further enable the exchange of the LexA DBD-MBP-VP64 fragment by any sequence of interest.
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:pDN80 was a gift from Barbara Di Ventura & Roland Eils (Addgene plasmid # 61349 ; http://n2t.net/addgene:61349 ; RRID:Addgene_61349)
For your References section:Engineering light-inducible nuclear localization signals for precise spatiotemporal control of protein dynamics in living cells. Niopek D, Benzinger D, Roensch J, Draebing T, Wehler P, Eils R, Di Ventura B. Nat Commun. 2014 Jul 14;5:4404. doi: 10.1038/ncomms5404. 10.1038/ncomms5404 PubMed 25019686