1. Integration of insert from expression vector pWD2-CpFBP1 into the C. reinhardtii chloroplast genome. (a) Schematic depicting the insert region of chloroplast expression vector
pWD2-CpFBP1 (top) and its integration into the region of the chloroplast (Cp) genome between the 5S rRNA gene and exon 5 of the psbA gene (middle and bottom). pWD2-CpFBP1
contains ~1.3-kb and 1.1-kb regions of homology to the Cp genome (nucleotides 45,892–47,242 and 47,349–48,407 for 5′ and 3′ regions, respectively; crosshatched boxes); a 1125-bp
C. reinhardtii FBP1 coding fragment (white box) flanked with 263 bp of psbD promoter/5′-UTR and 374 bp of psbA 3′-UTR (gray boxes); and a ~ 1.9 kb aadA cassette encoding SpecR
(white box) flanked by 652 bp of atpA promoter/5′-UTR and 436 bp of rbcL 3′UTR. The entire FBP1 + aadA gene cassette is 3.7 kb. Numbers below EcoRI sites in the schematic of the
Cp genome region indicate bp coordinates in the Cp genome.