pComb3HTT
(Plasmid #63900)

• Purpose: Backbone of pComb3H containing TT Fab (light and heavy chains) against tetanus toxin, used as phage display/expression control.
• Depositing Lab: Carlos Barbas
• Publication: Barbas et al Methods, A companion to Methods in Enzymology, 1995; 8(2): 94-103.

Backbone

• Vector backbone: pComb3H
• 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
• Tag / Fusion Protein:
  • 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: 5'-AAG ACA GCT ATC GCG ATT GCA G-3'
• 3′ sequencing primer: 5'-GCC CCC TTA TTA GCG TTT GCC ATC-3'

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

pComb3H is the second generation of pComb vector. Improvements over pComb3 include increased stability and introduction of a new cloning region utilizing asymmetrical SfiI restriction sites for directional cloning of full Fab, scFv, peptide and other proteins for phage display. The “TT” refers to the human Fab to tetanus toxin which can be used as a phage display and protein expression control. Soluble Fab can be expressed by removing the gene for the pIII phage fusion protein by SpeI/NheI digest. Sufficient soluble protein is also found in the periplasmic space as a result of proteolysis.