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pSANG14-3F
(Plasmid #39265)

Ordering

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

  • Vector backbone
    pSANG10-3F
  • Backbone manufacturer
    Addgene plasmids #39264
  • Backbone size (bp) 5438
  • Modifications to backbone
    The 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 type
    Bacterial Expression
  • Promoter T7
  • Tags / 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

  • Bacterial Resistance(s)
    Kanamycin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    Must be transformed into BL21(DE3) bacteria and grown at 30 degrees for antibody production.
  • Copy number
    Unknown

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ sequencing primer T7 promoter
  • 3′ sequencing primer T7 terminal primer
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

Bacterial expression of recombinant antibody fragments in the form of single chain Fvs (scFvs), dimerised through E. coli alkaline phosphatase

How to cite this plasmid ( Back to top)

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