pOTTC1080 - prRosa26v2 LSL FLAG-SpCas9
(Plasmid #113163)

• Purpose: A donor plasmid with homologous arms matching rat Rosa26 and expressing a Cre-dependent FLAG-tagged SpCas9
• Depositing Lab: Brandon Harvey
• Publication: Back et al Neuron. 2019 Feb 8. pii: S0896-6273(19)30062-5. doi: 10.1016/j.neuron.2019.01.035.

Backbone

• Vector backbone: pZDrRosa26
• Backbone manufacturer: Sigma
• Backbone size w/o insert (bp): 5000
• Total vector size (bp): 12075
• Modifications to backbone: extended left and right homologous arms to be longer (1kb from gRNA sites), amplified from rat genome.
• Vector type: Mammalian Expression, CRISPR

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: SpCas9
• Alt name: S. pyogenes Cas9 nuclease
• Alt name: Cas9
• Species: Synthetic
• Insert Size (bp): 4200
• Promoter: CAG
• Tag / Fusion Protein:
  • FLAG (N terminal on insert)

Cloning Information

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: M13-rev
• 3′ sequencing primer: M13-fwd

Resource Information

• Supplemental Documents:
  • pOTTC1080 - prRosa26v2 LSL FLAG-SpCas9
• A portion of this plasmid was derived from a plasmid made by: The insert was amplified from PX458, addgene 48138

Terms and Licenses

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

Depositor Comments

Please note: Addgene NGS is unable to fully resolve the CAG promoter sequence. Please refer to the depositor's sequence for this section of the plasmid.