pPKm-230
(Plasmid #90499)

• Purpose: pSIN - EF-1alpha - PIF3 - MTAD - IRES - PhyB - GBD, dual vector of PIF3-MTAD under EF-1 alpha promoter and PhyB-DBD under IRES promoter; See growth conditions below.
• Depositing Lab: Todd Coleman
• Publication: Kyriakakis et al ACS Synth Biol. 2018 Feb 16;7(2):706-717. doi: 10.1021/acssynbio.7b00413. Epub 2018 Jan 24.

Backbone

• Vector backbone: pSIN
• Vector type: Lentiviral

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Growth instructions: Pick a small colony after streaking or transforming, as plasmid is very unstable. We recommend screening several small colonies by restriction digest before storing immediately upon receipt.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: mito-tFD and mito-tFNR

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: unknown (unknown if destroyed)
• 3′ cloning site: unknown (unknown if destroyed)

Resource Information

• Supplemental Documents:
  • pPKm-230.ape
• Addgene Notes:
  • Addgene Diagnostic Digest

Terms and Licenses

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

Depositor Comments

Additional growth instructions: For larger cultures, the depositing lab minipreps a sample before midiprepping. Perhaps if you are just making glycerol stocks try growing 2 or maybe 3 large cultures and miniprepping a sample to check for the full sized plasmid. They also try to start with a small sized colony and are more stringent about not letting the culture overgrow.