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Biobrick vector contains antibiotic resistance marker and an origin of replication, and a number of restriction enzyme sites. We could replace the resistance marker with a resistance gene and a specific ori. Make a biobrick vector where we swap in MoClo (Modular Cloning( sites. MoClo allows simultaneous rather than just sequential operations/digestions.
- restriction enzymes to cut
- Ligase to stick together
- Phosphatase --> phosphate groups
Ligase links the groups together-
Issues with old way: the complementary ends on the plasmid could stick to eachother. So use 2 different restriction enzymes so the restriction sites are different for each end so it can't bind itself.
. Use selection. Treat with enzyme