Table 1. Plasmids used in this study.
| Plasmida | Promoter for GOI | GOI | Junction | Selectable Marker |
|---|---|---|---|---|
| pMO511b | PPSAD | CrVENUS-3FLAG | PH/R | APHVIII |
| pMO424b | PPSAD | LifeAct-CrVENUS-3FLAG | PH/R | APHVIII |
| pMO448 | PH/R | N/A | N/A | APHVIII |
| pMO449 | PH/R | CrVENUS-3FLAG | TAGCAT | APHVIII |
| pMO459 | PH/R | LifeAct-CrVENUS-3FLAG | TAGCAT | APHVIII |
| pMO470 | PH/R | CrVENUS-3FLAG | TGATAGCCAT | APHVIII |
| pMO471 | PH/R | CrVENUS-3FLAG | TAGCCAT | APHVIII |
| pMO480 | PH/R | CrVENUS-3FLAG-CVIMc | TAGCAT | APHVIII |
| pMO482 | PH/R | PRO1-CrVENUS-FLAG | TAGCAT | APHVIII |
| pMO483 | PH/R | IDA5-CrVENUS-3FLAG | TAGCAT | APHVIII |
| pMO484 | PH/R | VFL2-CrVENUS-3FLAG | TAGCAT | APHVIII |
| pMO488 | PH/R | CrVENUS | TAGCAT | APHVIII |
| pMO490 | PH/R | sfGFP-3FLAG | TAGCAT | APHVIII |
| pMO507 | PPSAD | CrVENUS-3FLAG | TAGCAT | APHVIII |
| pMO508 | PTUB2 | CrVENUS-3FLAG | TAGCAT | APHVIII |
| pMO515 | PH/R | PMH1-CrVENUS-3FLAG | TAGCAT | APHVIII |
| pMO518 | PH/R | CrVENUS-3FLAG | TRBCS2-PPSAD | APHVIII |
| pMO519 | PH/R | CrmCHERRY | TAGCAT | APHVIII |
| pMO520 | PH/R | CrmCHERRY-3FLAG | TAGCAT | APHVIII |
a
Except for pMO511 (note b and Figure 1A), pMO424 (note b), and pMO518 (Figure 1A), all plasmids listed have the general structure shown for pMO449 in Figure 1B.
b
In pMO511 (Figure 1A) and pMO424 (Avasthi et al. 2014), the selectable marker is upstream under the control of the PH/R promoter, and the GOI is downstream under the control of the PPSAD promoter. pMO424 is identical to pMO511 except that the Chlamydomonas-codon-optimized Lifeact-encoding sequence is fused at the N-terminus of CrVENUS-3FLAG.
c
CVIM is expected to function as a “CAAX-box” for prenylation.