pOpen-T4 PGD
(Plasmid #165550)

• Purpose: T4 pyrimidine DNA glycosylase; bifunctional DNA glycosylase with DNA N-glycosylase and AP lyase activities.
• 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• Publication: Open Enzyme Collection (unpublished)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: T4 PGD (T4 Endonuclease V)
• 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

T4 pyrimidine DNA glycosylase; bifunctional DNA glycosylase with DNA N-glycosylase and AP lyase activities. The N-glycosylase activity releases cis-syn cyclobutane pyrimidine dimers, including T^T, T^C and C^C, generating an AP site. The AP lyase activity cleaves an AP site via beta-elimination, creating a 1 nucleotide DNA gap with 3'-alpha, beta-unsaturated aldehyde and 5'-phosphate termini. Also known as T4 Endonuclease V. This enzyme in combination with other enzymes from E. coli (UDG, fpg, Endonuclease IV, Endonuclease VIII), Bst-DNA pol and Taq/Tth-DNA Ligase forms the proprietary NEB Enzyme mix preCR.

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/