pOpen-T7EI
(Plasmid #165540)

• Purpose: DNA Endonuclease; catalyzes cleavage of DNA mismatches and non-beta DNA structures (Holliday junctions and cruciform).
• 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• Publication: Open Enzyme Collection (unpublished)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: T7EI (T7 Endonuclease I)
• 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

DNA Endonuclease; catalyzes the cleavage of DNA mismatches and non-beta DNA structures including Holliday junctions and cruciforms, leaving 3'-OH and 5'-phosphate. Best at C mismatches and does not recognize all DNA mismatches. To a lesser extent, cleaves across a nick in dsDNA; Key enzyme for use in genome editing mutation detection workflows.

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/