pSc++
(Plasmid #129106)

• Purpose: *Please note the plasmid name has been updated. Please see Plasmid 155011: Sc++ (https://www.addgene.org/155011/). Mammalian expression vector encoding codon-optimized Sc++ Cas9 nuclease.
• Depositing Lab: Joseph Jacobson
• Publication: Robust Genome Editing of Single-Base PAM Targets with Engineered ScCas9 Variants (unpublished)

Backbone

• Vector backbone: Adapted from pCMV_BE4max_P2A_GFP
• Backbone size w/o insert (bp): 6049
• Total vector size (bp): 10174
• Modifications to backbone: Added WPRE
• Vector type: Mammalian Expression, CRISPR
• Selectable markers: GFP

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Sc++ CDS
• Species: Synthetic
• Insert Size (bp): 4125
• Mutation: I365A, G366D, I367K, H369L, T373S, T374G, Q379E, T1227K
• Promoter: EF1⍺
• Tag / Fusion Protein:
  • NLS-P2A-EGFP-NLS (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: EF-1α Forward
• 3′ sequencing primer: SV40pA-R

Resource Information

• Supplemental Documents:
  • pScPP.gb

Terms and Licenses

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

Depositor Comments

Please note the name and information of this plasmid have been updated to Plasmid 155011: Sc++ in the Chatterjee et al Nat Biotechnol. 2020 publication. Please see the new webpage: https://www.addgene.org/155011/