pSm-Nuclease
(Plasmid #132431)

• Purpose: Expresses Serratia marcescens nuclease in E. coli using T7 promoter
• Depositing Lab: Mark Arbing
• Publication: His6 tagged mCherry (unpublished)

Backbone

• Vector backbone: pET24a
• Backbone size w/o insert (bp): 5230
• Total vector size (bp): 5981
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: Serratia marcescens nuclease (codon optimized)
• Alt name: NucA (Uniprot: P13717)
• Species: Synthetic
• Insert Size (bp): 744
• Promoter: T7
• Tag / Fusion Protein:
  • None

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: T7
• 3′ sequencing primer: T7 terminator

Resource Information

• Supplemental Documents:
  • psm-nuclease-benzonase.gb

Terms and Licenses

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

Depositor Comments

Target gene is expressed from a T7 promoter so DE3 strain must be used for protein expression. The target is insoluble and can be refolded using the protocol from Friedhoff et al. A procedure for renaturation and purification of the extracellular Serratia marcescens nuclease from genetically engineered Escherichia coli. Protein Expr Purif. 1994 Feb;5(1):37-43.

The nuclease is similar to Benzonase in that it is a potent endonuclease capable of degrading single and double stranded RNA and DNA.