pAG32-3HA-3FLAG
(Plasmid #154081)

• Purpose: For C-terminal 3xHA-3xFLAG-protein tagging in S. pombe using hphMX4 as resistance marker.
• Depositing Lab: Cornelia Kilchert
• Publication: Kilchert et al Genome Res. 2020 Jul;30(7):1012-1026. doi: 10.1101/gr.257006.119. Epub 2020 Jun 18.

Backbone

• Vector backbone: pFA6a-hygR = pAG32
• Backbone size w/o insert (bp): 4160
• Total vector size (bp): 4606
• Modifications to backbone: insertion of 3xHA-3FLAG-TAG + ADH1 terminator
• Vector type: PCR-based tagging in yeast
• Selectable markers: Hygromycin

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: 3xHA-3xFLAG-hygR
• Insert Size (bp): 2154
• Tag / Fusion Protein:
  • 3xHA-3xFLAG (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SpeI (not destroyed)
• 3′ cloning site: BglII (not destroyed)
• 5′ sequencing primer: ScAdh1-term-rev

Resource Information

• Supplemental Documents:
  • pAG32_HA_flag.gbk
• A portion of this plasmid was derived from a plasmid made by: SpeI/BglII-restricted hphMX4 from pAG32 (Goldstein and McCusker (1999) Yeast 15: 1541-1553)

Terms and Licenses

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