pX459-HypaCas9-mR2-TUBA1B_sgRNA
(Plasmid #183888)

• Purpose: pX459V2.0-HypaCas9 plasmid with mRuby2 reporter and TUBA1B sgRNA for N-terminal tagging of alpha-tubulin in human cells.
• Depositing Lab: Alisa Piekny
• Publication: Husser et al Open Biol. 2022 Nov;12(11):220247. doi: 10.1098/rsob.220247. Epub 2022 Nov 23.

Backbone

• Vector backbone: pX459V2.0-HypaCas9-mRuby2
• Total vector size (bp): 9955
• 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): NEB Stable
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: TUBA1B sgRNA spacer
• Alt name: Tubulin alpha
• gRNA/shRNA sequence: GATGCACTCACGCTGCGGGA
• Species: H. sapiens (human)
• Entrez Gene: TUBA1B (a.k.a. K-ALPHA-1)
• Promoter: U6

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BbsI (destroyed during cloning)
• 3′ cloning site: BbsI (destroyed during cloning)
• 5′ sequencing primer: gagggcctatttcccatgattcc
• 3′ sequencing primer: ggccatttaccgtaagttatgtaacg

Resource Information

• Supplemental Documents:
  • pX459V2.0-HypaCas9-mRuby2-sgRNA_TUBA1B.gb

Terms and Licenses

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