pCas9-sgAAVS1-2
(Plasmid #129727)

• Purpose: Expressing SpCas9 and sgAAVS1-2
• Depositing Lab: Elina Ikonen
• Publication: Li et al Nat Methods. 2019 Aug 26. pii: 10.1038/s41592-019-0512-x. doi: 10.1038/s41592-019-0512-x.

Backbone

• Vector backbone: pGL3-basic
• Backbone manufacturer: Promega
• Backbone size w/o insert (bp): 2000
• Total vector size (bp): 9321
• Modifications to backbone: Only plasmid backbone for amplification in E.coli was used.
• Vector type: Mammalian Expression, CRISPR
• Selectable markers: Puromycin ; mCherry for FACS enrichment

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: SpCas9 and sgAAVS1-2
• gRNA/shRNA sequence: GTCACCAATCCTGTCCCTAG tgg
• Species: H. sapiens (human)
• Promoter: hU6
• Tag / Fusion Protein:
  • T2A-mCherry-P2A-Puro (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: KpnI (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: gactatcatatgcttaccgt
• 3′ sequencing primer: -

Resource Information

• Supplemental Documents:
  • pCas9-sgAAVS1-2 sgA2-1.gb
• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

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