pSANG10-3F
(Plasmid #39264)

• Purpose: (Empty Backbone)
• Depositing Lab: John McCafferty
• Publication: Martin et al BMC Biotechnol. 2006 Dec 7;6:46.

Backbone

• Vector backbone: pET26b(+)
• Backbone manufacturer: EMD Biosciences
• Backbone size (bp): 5438
• Modifications to backbone: pSANG10 was created by digesting pET26b(+) at the single XhoI site and at the BglI sites closest to the T7 promoter (by partial digestion) and replacing the fragment with a double stranded oligonucleotides compatible with these sites, using four annealed oligonucleotides (see Table 2 of manuscript for all oligonucleotide sequences). This introduced NcoI, PstI, NotI, HindIII restriction sites and a serine (pSANG10) immediately after the NotI site. The tri-FLAG epitope tag was inserted into pSANG10 at the HindIII site using two annealed oligonucleotides to create pSANG10-3F. 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 ; Bacterial expression, bacterial expression of recombinant antibody fragments in the form of single chain Fvs (scFvs)
• Promoter: T7
• Tags / Fusion Proteins:
  • 6xHis (C terminal on backbone)
  • tri-FLAG (C terminal on backbone)
  • pelB leader (N terminal on backbone)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• 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

Resource Information

• Supplemental Documents:
  • pSANG10-3F.gb
• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

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