pC1 Grx1-roCherry-mito
(Plasmid #141069)

• Purpose: Fluorescent biosensor for 2GSH/GSSG ratio with mitochondrial localization
• Depositing Lab: Dmitry Bilan
• Publication: Shokhina et al Redox Biol. 2019 Feb;21:101071. doi: 10.1016/j.redox.2018.101071. Epub 2018 Dec 7.

Backbone

• Vector backbone: pC1
• Backbone size w/o insert (bp): 3919
• Total vector size (bp): 5194
• Modifications to backbone: Grx1-roCherry was inserted into pC1 backbone, which was previously equipped with MLS1 and MLS2
• Vector type: Mammalian 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: Grx1-roCherry
• Species: Synthetic
• Insert Size (bp): 1275
• Mutation: Changed Alanine 150 to Cysteine, inserted Threonine after Cysteine 150, changed Serine 151 to Glutamate, changed Lysine 203 to Cysteine
• Promoter: CMV
• Tag / Fusion Protein:
  • dMTS (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: CCA TTG ACG TCA ATG GGA GTT
• 3′ sequencing primer: CAA GTT AAC AAC AAC AAT TGC AT

Resource Information

• Supplemental Documents:
  • pC1_dMTS Grx1-roCherry.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry