|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||39265||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Backbone manufacturerAddgene plasmids #39264
- Backbone size (bp) 5438
Modifications to backboneThe alkaline phosphatase encoding gene was inserted between the scFv and 6xHis tag in pSANG10-3F to create pSANG14-3F. The scFv encoding gene is sub-cloned at the NcoI/NotI site and the 5' HindIII site is available for insertion of peptide tags and fusion partners.
Vector typeBacterial Expression
- Promoter T7
/ Fusion Proteins
- pelB leader (N terminal on insert)
- Alkaline phosphatase (C terminal on insert)
- 6xHis (C terminal on insert)
- tri-FLAG (C terminal on insert)
Growth in Bacteria
Growth instructionsMust be transformed into BL21(DE3) bacteria and grown at 30 degrees for antibody production.
- Cloning method Restriction Enzyme
- 5′ sequencing primer T7 promoter
- 3′ sequencing primer T7 terminal primer (Common Sequencing Primers)
Bacterial expression of recombinant antibody fragments in the form of single chain Fvs (scFvs), dimerised through E. coli alkaline phosphatase
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:pSANG14-3F was a gift from John McCafferty (Addgene plasmid # 39265 ; http://n2t.net/addgene:39265 ; RRID:Addgene_39265)
For your References section:A simple vector system to improve performance and utilisation of recombinant antibodies. Martin CD, Rojas G, Mitchell JN, Vincent KJ, Wu J, McCafferty J, Schofield DJ. BMC Biotechnol. 2006 Dec 7;6:46. 10.1186/1472-6750-6-46 PubMed 17156422