pAAV-U6-sasgRNACMV- mCherry-WPREpA
(Plasmid #217015)

• Purpose: S. aureus sgRNAs backbone along with mCherry expression
• Depositing Lab: Nadav Ahituv
• Publication: Matharu et al Science. 2018 Dec 13. pii: science.aau0629. doi: 10.1126/science.aau0629.

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Backbone

• Vector backbone: pAAVCMV- sadCas9-VP64-pA
• Total vector size (bp): 4963
• Vector type: Mammalian Expression, AAV, 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: mCherry
• gRNA/shRNA sequence: None
• Species: Discosoma sea anemones
• Promoter: CMV

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: T7
• 3′ sequencing primer: T3

Resource Information

• A portion of this plasmid was derived from a plasmid made by: pAAV-U6-sasgRNACMV-mCherry-WPREpA was cloned by replacing the CMV-sadCas9-VP64-pA cassette in pAAVCMV-sadCas9-VP64-pA backbone with that of U6-sasgRNA-CMV-mCHerry-WPREpA cassette from pLTR-1120-MP177-U6-sasgRNA-mcherry (a kind gift from B. Huang at UCSF).

Terms and Licenses

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

Depositor Comments

S. aureus sgRNAs can be cloned using the In-Fusion HD-cloning kit (Clontech) into the Bst XI and Xho I
restriction enzyme sites.