pORANGE_Tubb3 GFP(Y39TAG) KI
(Plasmid #182678)

• Purpose: Expression of spCas9, gRNA targeting the end of Tubb3 gene and donor GFP(Y39TAG). Can be used for amber codon suppression and click chemistry labeling of endogenous beta-3 tubulin in mammalian cells.
• Depositing Lab: Ivana Nikić-Spiegel
• Publication: Arsic et al Nat Commun. 2022 Jan 14;13(1):314. doi: 10.1038/s41467-022-27956-y.

Backbone

• Vector backbone: pORANGE
• Vector type: Mammalian Expression

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: Tubb3-targeting gRNA and GFP(Y39TAG) donor sequence
• Alt name: Tubb3
• gRNA/shRNA sequence: GCTGCGAGCAACTTCACTT
• Species: M. musculus (mouse), R. norvegicus (rat)
• GenBank ID: NM_023279.3 NM_139254.2
• Entrez Gene: Tubb3 (a.k.a. 3200002H15Rik, M(beta)3, M(beta)6)
• Entrez Gene: Tubb3
• Promoter: U6 and chicken beta-actin promoter

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: aaggctgttagagagataattggaa
• 3′ sequencing primer: cgggccatttaccgtaagtt

Resource Information

• A portion of this plasmid was derived from a plasmid made by: This plasmid is based on the pORANGE Tubb3-GFP KI plasmid: https://www.addgene.org/131497/. We replaced WT GFP with the GFP(Y39TAG), all other sequences were unchanged.

Terms and Licenses

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