pUS2-SH231
(Plasmid #115150)

• Purpose: Cas9-GFP, dual gRNA expression vector for targeted integration of repair templates into safe harbor 231 locus in chromosome 4
• Depositing Labs: Raymond Monnat, Michael Phelps
• Publication: Pellenz et al Hum Gene Ther. 2019 Jul;30(7):814-828. doi: 10.1089/hum.2018.169. Epub 2019 Mar 28.

Backbone

• Vector backbone: custom
• Backbone size w/o insert (bp): 2739
• Vector type: Mammalian Expression, CRISPR

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: Dual SH231 gRNAs
• Alt name: Cas9-T2A-GFP
• gRNA/shRNA sequence: gcctcccccatagtaccat, Gatgtgctcactgagtctga
• Species: Streptococcus pyogenes
• Insert Size (bp): 6866
• Promoter: U6, CBh
• Tags / Fusion Proteins:
  • 3xFLAG-SV40NLS (N terminal on backbone)
  • GFP (C terminal on insert)
  • Nucleoplasmin NLS (C terminal on backbone)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: unknown

Resource Information

• Supplemental Documents:
  • pUS-SH231.gb

Terms and Licenses

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

Depositor Comments

Please visit https://www.biorxiv.org/content/early/2018/08/20/396390 for bioRxiv preprint.