pET-29b(+)_NSP4ss_SerM_NucA-His
(Plasmid #218190)

• Purpose: Bacterial expression of a periplasmically expressed His-tagged Serratia Marcescens NucA (Benzonase)
• Depositing Lab: Florence Pojer
• Publication: Homemade Benzonase (unpublished)

Backbone

• Vector backbone: pET-29b(+)
• Backbone manufacturer: Novagen
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): T7 Express Iq lysY
• Growth instructions: For expression, grow in your preferred BL21(DE3) or related strain. Induce with 0.5 mM IPTG at 18C overnight.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: nucA
• Species: Serratia marcescens
• Mutation: codon optimized ecoli residues - 22-266
• GenBank ID: AAA26560.1 P13717
• Promoter: T7
• Tags / Fusion Proteins:
  • NSP4 signal secretion peptide (N terminal on insert)
  • His (C terminal on insert)

Cloning Information

• Cloning method: Gene Synthesis
• 5′ sequencing primer: T7promoter
• 3′ sequencing primer: T7terminator

Resource Information

• Supplemental Documents:
  • 4_pET-29b(+)_NSP4ss_SerM_NucA-His.gbk

Terms and Licenses

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

Depositor Comments

Plasmid expresses a nuclease. Leaky expression of the nuclease may degrade extracted plasmid DNA over time. It is recommended that this plasmid be maintained in the provided host strain. If plasmid is extracted, use it immediately.