CMV-hCas9-T2A-mStrawberry
(Plasmid #187224)

• Purpose: Expresses human Cas9 nuclease and mStrawberry via T2A linker under control of a CMV promoter.
• Depositing Lab: Jennifer Mitchell
• Publication: Abatti et al Nucleic Acids Res. 2023 Sep 22:gkad734. doi: 10.1093/nar/gkad734.

Backbone

• Vector backbone: pcDNA3.3-TOPO
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 6390
• Total vector size (bp): 10533
• Modifications to backbone: Inserted T2A-mStrawberry at 3' terminal of MCS.
• Vector type: Mammalian Expression, CRISPR
• Selectable markers: Neomycin (select with G418)

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: Cas9
• Species: H. sapiens (human)
• Insert Size (bp): 4140
• GenBank ID:
• Promoter: CMV
• Tag / Fusion Protein:
  • Inserted T2A-mStrawberry at 3' terminal of MCS. (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: TAGAAGACACCGGGACCGAT
• 3′ sequencing primer: GGATTCTCCTCCACGTCACC

Resource Information

• Supplemental Documents:
  • hcas9-t2a-mstrawberry.gbk
• A portion of this plasmid was derived from a plasmid made by: hCas9 was subcloned from Addgene Plasmid #41815 (hCas9 from George Church)

Terms and Licenses

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