Intermediates and proposed constructs for the the 2 plasmid locking system
In this design, each of the construct has its own antibiotic resistance marker (independent of the plasmid backbone marker).
We decided to remove the extra antibiotic markers from the inserts. Then it looks like this
We had more than one way to build plasmid 1
Intermediates and proposed constructs for the the 3 plasmid locking system
We have designed the constructs for the 3 plasmid system, but we haven't planned to build it yet. Some of the parts built for the 2 plasmid system could be reused for the 3 plasmid system too.
Intermediates planned through PCR protocol
These parts are essentially same as the those that occur in the construction strategies that are shown above.