pET-6xHis-(pos15)GFP
(Plasmid #89248)

• Purpose: [Edit] Expresses GFP with +15 theoretical charge in E.Coli
• Depositing Lab: David Liu
• Publication: Thompson et al Chem Biol. 2012 Jul 27;19(7):831-43. doi: 10.1016/j.chembiol.2012.06.014.

Backbone

• Vector backbone: pET29
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5152
• Total vector size (bp): 5896
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Can also maintain in Mach1. For protein expression, transform into BL21 Star (DE3) cells. Induce at 0.6 OD with 0.5 mM IPTG final concentration. Growing overnight (16 hours) at 20-25 C.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: His6-(pos15)-GFP
• Species: Synthetic
• Insert Size (bp): 745
• Promoter: T7
• Tag / Fusion Protein:
  • His6 (N terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: T7 Promoter
• 3′ sequencing primer: T7 Terminator

Terms and Licenses

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