Description:

Recombineering plasmid designed for generating C-terminal CrVenusYFP-3xFLAG fusion proteins. Contains the counter-selection ccdB gene that is toxic to bacteria not expressing ccdA. The primers below can be used to amplify a cassette from the plasmid that excludes ccdB. A gene of interest can then be cloned into the cassette in-frame with CrVenusYFP-3xFLAG by recombineering. Forward primer (5'-3'): GGAGATCTGGGTGGCTCCG. Reverse primer (5'-3'): GAAGATCCTTTGATCTTTTCTACGGG. These primer sequences should be appended to the 3' end of gene-specific homology regions. Selection in E. coli: kanamycin. Selection in C. reinhardtii: paromomycin.

Bacterial Host Strain:

DB3.1

Selectable Marker:

Sm-r, ccdB-r