pSADuet148
(Plasmid #121910)

• Purpose: Charge-introduced NpuDnaBΔ283ΔC39 intein
• Depositing Lab: Hideo Iwai
• Publication: Beyer et al FEBS J. 2019 Oct 30. doi: 10.1111/febs.15113.

Backbone

• Vector backbone: pRSFDuet-1
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 3994
• Total vector size (bp): 4289
• Modifications to backbone: His6-GB1 tag, deletion of MCS-2
• 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: CI-NpuDnaBΔ290ΔC39 intein
• Species: Nostoc punctiforme
• Insert Size (bp): 295
• Mutation: Deletion of 290-residue endonuclease domain, I53K, P55K, E58K
• GenBank ID: CP001037.1 ACC81364.1
• Promoter: T7
• Tag / Fusion Protein:
  • His6-GB1 (N terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ cloning site: BamHI (unknown if destroyed)
• 3′ cloning site: HindIII (unknown if destroyed)
• 5′ sequencing primer: T7 Forward
• 3′ sequencing primer: T7 Reverse

Resource Information

• Supplemental Documents:
  • pSADuet148.gb

Terms and Licenses

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