pJK_proJK1_LGK
(Plasmid #81160)

• Purpose: Expresses levoglucosan kinase with promoter proJK1
• Depositing Lab: Timothy Whitehead
• Publication: Klesmith et al ACS Synth Biol. 2015 Sep 14.

Backbone

• Vector backbone: pJK
• Backbone manufacturer: pET29 derivative
• Backbone size w/o insert (bp): 3582
• Total vector size (bp): 4899
• Modifications to backbone: Removal of the T7 promoter, T7 operator, lacI gene. Addition of the constitutive promoter proJK1 between BglI and XbaI. LGK is between NdeI and XhoI.
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Rich media for plasmid production as the constitutive promoter will reduce production under minimal media.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Levoglucosan kinase wild-type
• Alt name: B3VI55_LIPST
• Species: Synthetic; Lipomyces starkeyi
• Insert Size (bp): 1317
• GenBank ID: ACE79748.1 EU751287.1
• Promoter: proJK1
• Tag / Fusion Protein:
  • His6x (C terminal on insert)

Cloning Information

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

Terms and Licenses

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