OmpA3-PNGaseF-TEV-His6
(Plasmid #114274)

• Purpose: Amidase removing almost all N-linked oligosaccharides from glycoproteins. This PNGase F construct can have its hexa-his removed.
• Depositing Lab: Martin Caffrey
• Publication: PNGase F construct (unpublished)

Backbone

• Vector backbone: pBluescript KS(-)
• Backbone manufacturer: Stratagene
• Backbone size w/o insert (bp): 3000
• Modifications to backbone: Modification of plasmid #40315: Insertion of a TEV cleavage site between the hexaHis and the PNGase sequence to remove hexa-His
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: DH5alpha (storage), BL21(DE3) (expression)
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Peptide: N-glycosidase F
• Alt name: PNGase F
• Species: Flavobacterium meningosepticum
• Promoter: T7
• Tags / Fusion Proteins:
  • OmpA3 leader sequence (for periplasmic localisation) (N terminal on insert)
  • hexa-Histidine tag with TEV cleavage site (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: unknown (unknown if destroyed)
• 3′ cloning site: unknown (unknown if destroyed)
• 5′ sequencing primer: unknown

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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