pOpen-lambda red operon
(Plasmid #165521)

• Purpose: Operon containing Exo, Bet, and Gam. To use the lambda red recombineering system to modify your target DNA.
• 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• Publication: Open Enzyme Collection (unpublished)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: lambda red operon
• 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

Operon containing Exo, Bet, and Gam. To use the lambda red recombineering system to modify your target DNA (including E. coli genome), electroporate a linear donor DNA substrate (either dsDNA or ssDNA) into E. coli expressing the lambda red enzymes. These enzymes then catalyze the homologous recombination of the substrate with the target DNA sequence (mutagenesis occurs in vivo). ~50 nucleotides of homology are required to the target site for recombination.

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/