PurposeExpression and secretion of anti-GFP FLIPPER-bodies, small protein probes for correlated microscopy. FLIPPER-bodies are the fusion of a nanobody with a fluorescent protein and a peroxidase.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||112157||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 5371
- Total vector size (bp) 7528
Vector typeMammalian Expression
Selectable markersNeomycin (select with G418)
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert nameanti-GFP FLIPPER-body
Insert Size (bp)2157
- Promoter CMV
/ Fusion Proteins
- His (N terminal on insert)
- Thrombin cleavage site (N terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site HindIII (not destroyed)
- 3′ cloning site AgeI (not destroyed)
- 5′ sequencing primer TAATACGACTCACTATAGGGA
- 3′ sequencing primer TGGCAACTAGAAGGCACAGT (Common Sequencing Primers)
A portion of this plasmid was derived from a plasmid made bymCherry was directly derived from the Roger Tsien lab (Shaner et al. 2004) and is available via Addgene. Nanobody against GFP was from Addgene. HRP was amplified from FLIPPER (Kuipers et al. 2015), the original cDNA was provided by R.S. Lewis (Luik et al. 2006).
Terms and Licenses
- Not Available to Industry
Article Citing this Plasmid
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:pcDNA3.1 SP-His-mCherry-HRP-VhHGFP was a gift from Ben Giepmans (Addgene plasmid # 112157 ; http://n2t.net/addgene:112157 ; RRID:Addgene_112157)