pM9-BetaGal-8C
(Plasmid #172489)

• Purpose: Plasmid encoding E. coli BetaGalactosidase with an N-terminal M9 nuclear localization sequence
• Depositing Lab: Siegfried Musser
• Publication: Chowdhury et al Nat Cell Biol. 2022 Jan;24(1):112-122. doi: 10.1038/s41556-021-00815-6. Epub 2022 Jan 10.

Backbone

• Vector backbone: pET30a
• Backbone manufacturer: Novagen (EMD Millipore)
• Backbone size w/o insert (bp): 5422
• Total vector size (bp): 8617
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: M9-BetaGal-8C
• Alt name: BetaGal
• Species: Escherichia coli
• Insert Size (bp): 3363
• Mutation: BetaGal has two cysteine mutations (C77A, C1022S), to reduce the number of maleimide reactive sites, and two enzymatic mutations (E538Q, W999L), to reduce Kcat and sugar binding affinity.
• GenBank ID: WP_032298557.1 AAH71945.1
• Promoter: T7
• Tags / Fusion Proteins:
  • 6xHis (N terminal on insert)
  • M9 nuclear localization sequence (from HNRNPA1) (N terminal on insert)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: T7 Promoter primer
• 3′ sequencing primer: T7 Terminator primer

Resource Information

• Supplemental Documents:
  • pM9-BetaGal-8C.gbk
• A portion of this plasmid was derived from a plasmid made by: The M9-BetaGal parent plasmid was a gift of Larry Gerace, and first reported in Lyman, et al. (2002) J. Cell Biol. 159:55-67. doi: 10.1083/jcb.200204163

Terms and Licenses

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