pET21-pelB-LaG-29
(Plasmid #172753)

• Purpose: Bacterial expression of HIS-tagged anti-GFP nanobody LaG-29; HIS tag preceded by HRV 3C/PreScission cleavage site.
• Depositing Lab: Michael Rout
• Publication: Fridy et al Nat Methods. 2014 Dec;11(12):1253-60. doi: 10.1038/nmeth.3170. Epub 2014 Nov 2.

Backbone

• Vector backbone: pET21a(+)
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5443
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: LaG-29
• Species: Lama glama
• Insert Size (bp): 473
• Promoter: T7
• Tags / Fusion Proteins:
  • pelB leader for periplasmic secretion (N terminal on insert)
  • 6xHIS (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: T7 Term

Terms and Licenses

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

Depositor Comments

Insert codon-optimized for E. coli.