pAraH6HATT
(Plasmid #63901)

• Purpose: For expression of soluble scFv/Fab fragments in bacteria from genes selected using the pComb3H system, contains TT Fab (light and heavy chains) against tetanus toxin, used as 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: pAra
• Backbone size w/o insert (bp): 3500
• Total vector size (bp): 4900
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol, 25 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: araBAD
• Tags / Fusion Proteins:
  • 6xHis (N terminal on backbone)
  • HA tag (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'-AGA AGC GTA GTC CGG AAC GTC-3'

Terms and Licenses

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

Depositor Comments

Provided with the pComb3H system for arabinose induced expression of soluble protein with detection tags. The “TT” refers to the human Fab to tetanus toxin which can be used as a protein expression control. Alternatively, one could upgrade to the pComb3X system, also available on Addgene, which already contains tags.

Select reference:

Cary, S.P.; Lee, J.; Wagenknecht, R.; Silverman, G.J. (2000) Characterization of Superantigen-Induced Clonal Deletion with a Novel Clan III-Restricted Avian Monoclonal Antibody: Exploiting Evolutionary Distance to Create Antibodies Specific for a Conserved VH Region Surface. J. Immun., 164, 4730-4741.