pOpen-DNA Topoisomerase 1B Vaccinia Virus
(Plasmid #165510)

• Purpose: Used in TOPO cloning. Recognizes the DNA sequence 5'-(C/T)CCTT-3' and digests double stranded DNA at this sequence.
• 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• Publication: Open Enzyme Collection (unpublished)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: DNA Topoisomerase 1B Vaccinia Virus
• 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 TOPO cloning. Recognizes the DNA sequence 5'-(C/T)CCTT-3' and digests double stranded DNA at this sequence. The energy from this breakage is stored as a covalent bond between the DNA and the topoisomerase. If a 5' hydroxyl group from a different DNA strand comes along, it can attack this covalent bond thus joining the two DNA strands and releasing topoisomerase.

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/