pOpen-Endonuclease VIII E. coli
(Plasmid #165561)

• Purpose: Bifunctional DNA glycosylase with DNA N-glycosylase and AP lyase activities; The N-glycosylase activity releases damaged pyrimidines, including thymine glycol and uracil glycol. The AP lyase activity cleaves DNA phosphodiester backbone at AP sites via beta and delta-elimination, creating a 1 nucleotide DNA gap with 5' and 3' phosphate termini.
• 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• Publication: Open Enzyme Collection (unpublished)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: Endonuclease VIII E. coli
• 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

Used in the preCR mix from NEB; Bifunctional DNA glycosylase with DNA N-glycosylase and AP lyase activities; The N-glycosylase activity releases damaged pyrimidines, including thymine glycol and uracil glycol. The AP lyase activity cleaves DNA phosphodiester backbone at AP sites via beta and delta-elimination, creating a 1 nucleotide DNA gap with 5' and 3' phosphate termini.

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/