Figure 8. Cloning efficiency of level 1 and 2 constructs.

(A) Assembled transcription units are schematically represented as boxes annotated with the name of the CDS they contain. Transcription units shown in grey were cloned in the previous step (in construct cL2-6). The respective cloning position of each transcription unit is indicated on the top. For level 2 constructs, the end-linker is shown as a circle. The number of colonies obtained per transformation is shown by color type, with the first number corresponding to the expected correct constructs (for cL2-6, wrong clones could be either red or green). (B) Plates from transformation of constructs for level 2-1 (cL2-2, cL2-4 and cL2-5) and level 2i-1 (cL2-6). Since level 2-1 cloning uses red/white selection, the correct constructs should be white, while colonies containing the original destination vector construct should be red. Level 2i-1 uses a blue/red selection, with colonies containing correct constructs expected to be blue, whereas incorrect ones can be red or green (contain both the canthaxanthin operon and the lacZα fragment).