This part is composed of two mutated Flipase Recognition Targets (FRT') flanking the Chloramphenicol resistance gene. The mutation removes the XbaI restriction site from the canonical FRT sequence. It is similar to BBa_J61025 but without the scar between the first FRT' and CmR.
The two FRT' sequences allow the excision of the flanked sequence by the Flipase recombinase.
It's been tested with success for resistance to Chloramphenicol and excision per the pINDEL plasmid.
The pINDEL plasmid is composed of 2 functional units:
- the IN function which is composed of the gam, exo and bet genes coding for the lambda Red recombinase system
- the DEL function which is based on the flp gene encoding the FLP site-specific recombinase and expressed at 42°C
Another feature of pINDEL is that its replication origin is not functional at 42°C due to the thermosensitivity of the RepA101Ts protein. This protein initiates replication at the ori site in permissive conditions (30°C) and is inactive at 42°C.
The deletion function and the thermosensitivity of the plasmid both could be shown to be working as intended (see functional parameters). The insertion function couldn't be properly tested