Description:

FDX1 (Cre14.g626700) gDNA coding sequence, spanning the start codon to the triplet just before the stop codon, had been previously cloned in frame into pLM005 by Gibson assembly. That plasmid was then reengineered by restriction digestion and T4 ligation to add in-frame immediately after the 3xFLAG a synthetic fragment containing three copies of the sequence coding for the 15 C-terminal amino acids of SAGA2 (Cre 09.g394621), interspersed with a short flexible linker (GGGGGS). Transformed into C. reinhardtii strain cMJ030 with carrier DNA to produce MUT-5648. Selection in E. coli: ampicillin. Selection in C. reinhardtii: paromomycin.

Bacterial Host Strain:

DH5 alpha