pSpCas9n(BB)-2A-Puro (PX462) V2.0 Stx5DL-C1
(Plasmid #165085)

• Purpose: gRNA 1 of pair C for Stx5L-specific knockout
• Depositing Lab: Geert van den Bogaart
• Publication: Linders et al Nat Commun. 2021 Oct 28;12(1):6227. doi: 10.1038/s41467-021-26534-y.

Backbone

• Vector backbone: PX462
• Total vector size (bp): 9178
• Vector type: Mammalian Expression, CRISPR
• Selectable markers: Puromycin

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: hSpCas9n-2A-Puro
• Alt name: SpCas9n-2A-Puro V2.0
• Alt name: Cas9n
• Alt name: PX462 V2.0
• gRNA/shRNA sequence: TAACC TCGGA CTGTT GTGGA
• Species: Synthetic
• Promoter: CMV
• Tags / Fusion Proteins:
  • 3XFLAG (N terminal on insert)
  • 2A-Puro (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BbsI (unknown if destroyed)
• 3′ cloning site: BbsI (unknown if destroyed)
• 5′ sequencing primer: U6-fwd
• 3′ sequencing primer: U6-rev

Resource Information

• Supplemental Documents:
  • pSpCas9n(BB)-2A-Puro (PX462) v2_0 stx5DL-C1.gbk
• A portion of this plasmid was derived from a plasmid made by: Backbone from Addgene https://www.addgene.org/62987/

Terms and Licenses

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