pET-scFv-T
(Plasmid #67843)

• Purpose: pET-30-based vector dedicated to efficient scFv expression, which circumvents the problem of in-frame amber codons in scFvs.
• Depositing Lab: Joanna Bereta
• Publication: Ossysek et al Immunol Lett. 2015 Jul 25. pii: S0165-2478(15)30011-0. doi: 10.1016/j.imlet.2015.07.005.

Backbone

• Vector backbone: modified pET-30a
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5260
• Total vector size (bp): 5772
• Modifications to backbone: modified MCS, pelB leader sequence was introduced, addittional in frame His-tag was introduced
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Top10
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: antibody 1D8 light kappa chain
• Species: H. sapiens (human)
• Insert Size (bp): 512
• Promoter: T7 promoter
• Tag / Fusion Protein:
  • His-tag (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NcoI (not destroyed)
• 3′ cloning site: NotI (not destroyed)
• 5′ sequencing primer: TAATACGACTCACTATAGG
• 3′ sequencing primer: GCTAGTTATTGCTCAGCGG

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The antibody 1D8 light kappa chain was derived from Tomlinson I+J phagemid library.

Terms and Licenses

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