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Addgene

pComb3XTT
(Plasmid #63891)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 63891 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pComb3X
  • Backbone size w/o insert (bp) 3300
  • Total vector size (bp) 4700
  • Vector type
    Bacterial Expression ; phage display

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    XL1 Blue
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    TT Fab
  • Alt name
    human Fab to tetanus toxin
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    1400
  • Promoter lacZ
  • Tags / Fusion Proteins
    • 6xHis (C terminal on backbone)
    • HA tag (C terminal on backbone)
    • geneIII (C terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site SfiI (not destroyed)
  • 3′ cloning site SfiI (not destroyed)
  • 5′ sequencing primer AAG ACA GCT ATC GCG ATT GCA G
  • 3′ sequencing primer GCC CCC TTA TTA GCG TTT GCC ATC
  • (Common Sequencing Primers)

Resource Information

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

pComb3X is the newest of the pComb vectors. Improvements over pComb3 include increased stability and introduction of an asymmetric SfiI cassette for directional cloning of full Fab, scFv, peptide and other protein for phage display. 6xHis and HA tags allow for purification and detection. An amber stop codon was introduced to turn-off expression of the pIII fusion protein by switching to a non-supressor strain of E. coli allowing production of soluble protein without subcloning. Alternatively, the gene for phage protein pIII can be removed by SpeI/NheI digest. The “TT” refers to the human Fab to tetanus toxin which can be used as a template to amplify the kappa light chain constant region and Fd for construction of chimeric antibody libraries as described in:

Barbas, C. F., III; Burton, D. R.; Scott, J.K., Silverman, G.J. Eds. (2001) Phage Display: A Laboratory Manual; Cold Spring Harbor Laboratory Press: Cold Spring Harbor, New York

It can also be used as a phage display/protein expression control.

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:

    pComb3XTT was a gift from Carlos Barbas (Addgene plasmid # 63891 ; http://n2t.net/addgene:63891 ; RRID:Addgene_63891)
  • For your References section:

    Methods for the generation of chicken monoclonal antibody fragments by phage display. Andris-Widhopf J, Rader C, Steinberger P, Fuller R, Barbas CF 3rd. J Immunol Methods. 2000 Aug 28;242(1-2):159-81. 10.1016/s0022-1759(00)00221-0 PubMed 10986398