pOpen-T5gene12/13
(Plasmid #165559)

• Purpose: Double-stranded DNA specific exonuclease and single-stranded DNA endonuclease. Initiates at the 5' termini of linear or nicked double-stranded DNA. Cleaves linear or nicked double-stranded DNA in the 5' to 3' direction.
• Depositing Labs: Drew Endy, Jennifer Molloy, FreeGenes Project
• Publication: Open Enzyme Collection (unpublished)

Backbone

• Vector backbone: pBuild
• Vector type: Synthetic Biology

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: T5 Exonuclease
• Species: Other

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: AATG (unknown if destroyed)
• 3′ cloning site: GCTT (unknown if destroyed)
• 5′ sequencing primer: M13 forward
• 3′ sequencing primer: M13 reverse

Terms and Licenses

• Academic/Nonprofit Terms:
  • OpenMTA
• Industry Terms:
  • OpenMTA

Depositor Comments

Double-stranded DNA specific exonuclease and single-stranded DNA endonuclease. Initiates at the 5' termini of linear or nicked double-stranded DNA. Cleaves linear or nicked double-stranded DNA in the 5' to 3' direction. Ideal for removal of incomplete ligation products from ligated circular double-stranded DNA, degradation of denatured DNA from alkaline-based plasmid purification methods for improved DNA cloning, and degradation of contaminating linear and nicked DNA in plasmid samples.

The plasmids in the Open Enzyme collection contain genetic parts and are not functional by themselves. Researchers can clone these enzymes into an expression vector of their choice and then transform into E. coli bacteria for expression and purification. Additional information and protocols can be found at https://stanford.freegenes.org/