(559) pAAV EF-1a KRAB-SadCas9 U6-gRNA
(Plasmid #163024)

• Purpose: Expression of CRISPRi with U6-gRNA (Bsa1 sites)
• Depositing Lab: Tonia Rex
• Publication: Backstrom et al Mol Ther Methods Clin Dev. 2020 Sep 6;19:139-148. doi: 10.1016/j.omtm.2020.09.001. eCollection 2020 Dec 11.

Backbone

• Vector backbone: AAV2
• Backbone manufacturer: Cell Biolabs
• Backbone size w/o insert (bp): 3500
• Total vector size (bp): 7188
• Vector type: Mammalian Expression, AAV

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Growth instructions: 34-35 degrees is ideal
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Mammalian codon-optimized SadCas9
• Alt name: nuclease deactivated SauCas9
• Species: S. aureus
• Insert Size (bp): 3681
• Promoter: hEF-1a
• Tags / Fusion Proteins:
  • KRAB (N terminal on insert)
  • myc NLS (C terminal on insert)
  • SV40 polyA (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoR1 (not destroyed)
• 3′ cloning site: Kpn1 (not destroyed)
• 5′ sequencing primer: Mlu-EF1-Seq-S
• 3′ sequencing primer: Add-Avr2-A

Resource Information

• Supplemental Documents:
  • (559) pAAV EF-1a KRAB-SadCas9 U6-gRNA.gbk
• A portion of this plasmid was derived from a plasmid made by: Keith Joung (S. aureus dCas9 from Addgene #70703).

Terms and Licenses

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