pGFPuv carries the“cycle 3”variant of GFP described by Crameri et al. (1). This gene was cloned between the two MCSs of the pUC19 derivative pPD16.43 (2). The GFPuv gene canbe easily excised from pGFPuv. Alternatively, the GFPuv coding sequence can be amplified by PCR. The GFPuv gene was inserted in frame with the lacZ initiation codon from pUC19 so that a β-galactosidase-GFPuv fusionprotein is expressed from the lac promoter in E. coli . Note, however, that if you excise the GFPuv coding sequence using a restriction site in the 5' MCS, the resulting fragment will encode the native (i.e.,non-fusion) GFPuv protein. The pUC backbone of pGFPuv provides a high copy number origin of replication and ampicillin resistance gene for propagation in E. coli.