pOpen-T5gene122
(Plasmid #165544)

• Purpose: Possesses two enzymatic activities: DNA synthesis (polymerase) and exonucleolytic activity that degrades ssDNA in the 3'-5' direction for proofreading purposes.
• 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• Publication: Open Enzyme Collection (unpublished)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 DNA Polymerase
• 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

Replicates viral genomic DNA. Possesses two enzymatic activities: DNA synthesis (polymerase) and an exonucleolytic activity that degrades single-stranded DNA in the 3'-5' direction for proofreading purposes. The DNA synthesis very likely occurs by strand displacement.

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/