(517) SB KRAB-SadCas9 U6-gStop
(Plasmid #163023)

• Purpose: Sleeping Beauty TET-On expression of CRISPRi with gSTOP gRNA
• 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: Sleeping Beauty TET-On
• Backbone manufacturer: Eric Kowarz (Addgene plasmid # 60509)
• Backbone size w/o insert (bp): 6100
• Total vector size (bp): 9562
• Vector type: Mammalian Expression ; Transposon
• Selectable markers: Neomycin (select with G418)

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
• Species: S. aureus
• Insert Size (bp): 3483
• Promoter: Tet-On
• Tags / Fusion Proteins:
  • KRAB (N terminal on insert)
  • myc NLS (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoR1 (not destroyed)
• 3′ cloning site: Not1 (not destroyed)
• 5′ sequencing primer: SB-TRE-S
• 3′ sequencing primer: SVpa-Seq-A

Resource Information

• Supplemental Documents:
  • (517) SB KRAB-SadCas9 U6-gStop.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