pPK-352
(Plasmid #157922)

• Purpose: pcDNA-CMV-PCYA-IRES-HO1-P2A-FD-P2A-FNR
• Depositing Lab: Phillip Kyriakakis
• Publication: Kyriakakis et al J Vis Exp. 2021 Jan 12;(167). doi: 10.3791/61914.

Backbone

• Vector backbone: pcDNA3.1
• Vector type: Mammalian Expression, Synthetic Biology ; Optogenetics

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): 10-beta
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: tPcyA
• Alt name: Phycocyanobilin:ferredoxin oxidoreductase
• Species: S. cerevisiae (budding yeast); Thermosynechococcus elongatus
• Insert Size (bp): 799
• Promoter: CMV
• Tag / Fusion Protein:
  • Mitochondrial Targeting sequence (N terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: XBA (not destroyed)
• 5′ sequencing primer: T7 promoter
• 3′ sequencing primer: SP6 promoter

Gene/Insert 2

• Gene/Insert name: IRES
• Alt name: Internal Ribosomal Entry Site
• Insert Size (bp): 627

Gene/Insert 3

• Gene/Insert name: HO1
• Alt name: Heme Oxygenase
• Species: Thermosynechococcus elongatus
• Insert Size (bp): 813
• Tag / Fusion Protein:
  • Mitochondrial Targeting Sequence (N terminal on insert)

Gene/Insert 4

• Gene/Insert name: FD
• Alt name: Ferredoxin
• Species: Thermosynechococcus elongatus
• Insert Size (bp): 477
• Tag / Fusion Protein:
  • Mitochondrial Targeting Sequence and V5 tag (N terminal on insert)

Gene/Insert 5

• Gene/Insert name: FNR
• Alt name: Ferredoxin-NADP reductase
• Species: Thermosynechococcus elongatus
• Insert Size (bp): 1290
• Tag / Fusion Protein:
  • Mitochondrial Targeting Sequence and HIS tag (N terminal on insert)

Resource Information

• Supplemental Documents:
  • 352 - pcDNA-CMV-PCYA-IRES-HO1-FD-FNR.gb

Terms and Licenses

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