pBabe(puro)-Omi-mCherry
(Plasmid #48685)

• Purpose: Expresses fusion protein in mitochondria inter-membrane space, and is released upon MOMP.
• Depositing Lab: Douglas Green
• Publication: Tait et al Dev Cell. 2010 May 18;18(5):802-13.

Backbone

• Vector backbone: pBabe (puro)
• Backbone size w/o insert (bp): 5100
• Modifications to backbone: mCherry cloned via 5’BamH1/3’EcoRI restriction digest into pBabe puro. Human Omi (aka HtrA2) cloned via PCR/restriction digest using Omi-TC vector as template and primers with BglII restriction sites. PCR product was cut with BglII and cloned into pBabe mCherry via the BamHI site.
• Vector type: Mammalian Expression, Retroviral
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): Stbl2
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Omi
• Alt name: HTRA2
• Alt name: HtrA serine peptidase 2
• Alt name: PARK13
• Species: H. sapiens (human)
• GenBank ID: NM_013247.4
• Entrez Gene: HTRA2 (a.k.a. MGCA8, OMI, PARK13, PRSS25)
• Promoter: LTR
• Tag / Fusion Protein:
  • mCherry (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (destroyed during cloning)
• 3′ cloning site: BamHI (destroyed during cloning)
• 5′ sequencing primer: 5'-ctttatccagccctcac
• 3′ sequencing primer: 3'-accctaactgacacacattcc

Terms and Licenses

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