pBZ191-pU6-sgBFP-CBh-sv40NLS-Cas9-NLS-T2A-mCherry
(Plasmid #170183)

• Purpose: Expression vector for a sgRNA against BFP and spCas9 linked to mCherry via a T2A peptide
• Depositing Lab: Hunter Fraser
• Publication: Zhao et al CRISPR J. 2022 Jan 24. doi: 10.1089/crispr.2021.0065.

Backbone

• Vector backbone: pU6-(BbsI)_CBh-sv40NLS-Cas9-NLS T2A-mCherry
• Backbone manufacturer: Fraser lab
• Modifications to backbone: sgBFP was inserted by golden gate cloning into the vector backbone pU6-(BbsI)_CBh-sv40NLS-Cas9-NLS T2A-mCherry.
• Vector type: Mammalian Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: sgBFP
• Alt name: guide RNA against BFP
• Insert Size (bp): 20
• Promoter: hU6

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BbsI (destroyed during cloning)
• 3′ cloning site: BbsI (destroyed during cloning)
• 5′ sequencing primer: 5'-GACTATCATATGCTTACCGT-3'

Resource Information

• Supplemental Documents:
  • pbz191-pu6-sgbfp-cbh-sv40nls-cas9-nls-t2a-mcherry.gb

Terms and Licenses

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