pET29NK-LGK/mGFPmut3
(Plasmid #81168)

• Purpose: GFP fusion to levoglucosan kinase
• Depositing Lab: Timothy Whitehead
• Publication: Wrenbeck et al ACS Synth Biol. 2019 Mar 15;8(3):474-481. doi: 10.1021/acssynbio.8b00486. Epub 2019 Feb 8.

Backbone

• Vector backbone: pET29NK
• Backbone manufacturer: pET29b Derivative
• Backbone size w/o insert (bp): 5961
• Total vector size (bp): 7278
• Modifications to backbone: After the XhoI site, mGFPmut3 is cloned in-frame. After mGFPmut3 is the His6x tag, stop codon and then the BbvCI nicking site.
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• 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: T7
• Tags / Fusion Proteins:
  • mGFPmut3 (C terminal on backbone)
  • His6x (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: T7Term

Terms and Licenses

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