Ligations of vector with the Gateway C1 cassette, containing the suicide ccdB box with ccdB and chloramphenicol genes and attR sites (Invitrogen, Product No. 11828-019), were transformed into the DB3.1 Escherichia coli strain and positive clones were selected on chloramphenicol plates. Correct orientation of the cassette was verified by restriction digests and sequencing, resulting in the EGFP–pDEST vector.