pfMDH_K295A
(Plasmid #67726)

• Purpose: Expresses K295A mutant of mannitol 2-dehydrogenase from Pseudomonas fluorescens, codon optimized for expression in E. coli
• Depositing Lab: Justin Siegel
• Publication: Lucas et al Protein Sci. 2015 Jun;24(6):936-45. doi: 10.1002/pro.2669. Epub 2015 Apr 2.

Backbone

• Vector backbone: pET29b(+)
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5232
• Total vector size (bp): 6714
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH10B
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Mannitol 2-Dehydrogenase from Pseudomonas fluorescens
• Alt name: pfMDH
• Alt name: mtlD
• Species: Pseudomonas fluorescens
• Insert Size (bp): 1485
• Mutation: Changed lysine 295 to alanine
• GenBank ID: AAC04472.1
• Promoter: T7
• Tag / Fusion Protein:
  • 6x-His tag (C terminal on backbone)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: T7
• 3′ sequencing primer: T7term

Terms and Licenses

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