pFA6a-yomRuby3-SpHIS5
(Plasmid #204287)

• Purpose: For C-terminal tagging using yeast optimized mRuby3
• Depositing Lab: Linda Huang
• Publication: Seitz et al Mol Biol Cell. 2023 Sep 1;34(10):ar98. doi: 10.1091/mbc.E23-03-0096. Epub 2023 Jul 12.

Backbone

• Vector backbone: pFA-Envy-SpHIS5 (Addgene plasmid 60782)
• Backbone size w/o insert (bp): 4021
• Total vector size (bp): 4969
• Modifications to backbone: We replaced the Envy fluorescent marker with yomRuby3, which we had synthesized. We aslo added the meiotic high capacity terminator from the DIT1 gene at the 3' end
• Vector type: Yeast Expression
• Selectable markers: S pombe his5

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: pFA6-yomRuby3-SpHIS5
• Alt name: yomRuby3 for C-terminal tagging
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 1057
• Promoter: none
• Tag / Fusion Protein:
  • yomRuby3 (C terminal on backbone)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: SP6
• 3′ sequencing primer: GGG TAT TCT GGG CCT CCA TGT C

Resource Information

• Supplemental Documents:
  • pFA6a-yomRuby3-SpHIS5.gb
• A portion of this plasmid was derived from a plasmid made by: We synthesized yomRuby3 (Twist Bioscience)

Terms and Licenses

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