pET30a(+)-PPiase-C-Surmine
(Plasmid #261422)

• Purpose: Expresses PPiase with C-terminal Surmine (+ HisTag in E. coli), kanamycin 50 µg/ml, PPiase = inorganic pyrophosphatase (increases RNA yield and helps transcription by removing PP → 2 Pi)
• Depositing Lab: Andrei Lupas
• Depositing Organization: Max Planck Institute for Biology
  Located in Germany
• Publication: FluoroPURE Plasmids (unpublished)

Backbone

• Vector backbone: pET30a(+)
• Backbone manufacturer: Alexandra Waldherr (Max Planck), BioCat (Heidelberg, Germany)
• Backbone size w/o insert (bp): 5225
• Total vector size (bp): 6845
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: expresses well in BL21(DE3) (0.1-1 mM IPTG) O/N 18°C to 4 h 37°C
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: PPiase
• Alt name: Inorganic Pyrophospatase
• Alt name: PPA1
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 1620
• Entrez Gene: IPP1 (a.k.a. YBR011C, PPA1)
• Promoter: T7
• Tag / Fusion Protein:
  • Fluorescent protein (FP) and HIS-tag: C-terminal ...TEV-FP-His construct (C terminal on insert)

Cloning Information

• Cloning method: Gene Synthesis
• 5′ sequencing primer: GAAATTAATACGACTCACTATAGG

Terms and Licenses

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

Depositor Comments

This plasmid construct cannot be used for commercial purposes. Academic use please credit Alexandra Waldherr :)

Benchling link to our map: https://benchling.com/s/seq-1tHxFYqtGpHlhwilaXKS?item_id=seq_WjBHtAplSo&m=slm-0Bqt7Pfg1nkqSAi5eeTt