Table 1 Plasmids used in this study
PlasmidaPromoter for GOIGOIJunctionSelectable Marker
pMO511bPPSADCrVENUS-3FLAGPH/RAPHVIII
pMO424bPPSADLifeAct-CrVENUS-3FLAGPH/RAPHVIII
pMO448PH/RN/AN/AAPHVIII
pMO449PH/RCrVENUS-3FLAGTAGCATAPHVIII
pMO459PH/RLifeAct-CrVENUS-3FLAGTAGCATAPHVIII
pMO470PH/RCrVENUS-3FLAGTGATAGCCATAPHVIII
pMO471PH/RCrVENUS-3FLAGTAGCCATAPHVIII
pMO480PH/RCrVENUS-3FLAG-CVIMcTAGCATAPHVIII
pMO482PH/RPRO1-CrVENUS-FLAGTAGCATAPHVIII
pMO483PH/RIDA5-CrVENUS-3FLAGTAGCATAPHVIII
pMO484PH/RVFL2-CrVENUS-3FLAGTAGCATAPHVIII
pMO488PH/RCrVENUSTAGCATAPHVIII
pMO490PH/RsfGFP-3FLAGTAGCATAPHVIII
pMO507PPSADCrVENUS-3FLAGTAGCATAPHVIII
pMO508PTUB2CrVENUS-3FLAGTAGCATAPHVIII
pMO515PH/RPMH1-CrVENUS-3FLAGTAGCATAPHVIII
pMO518PH/RCrVENUS-3FLAGTRBCS2-PPSADAPHVIII
pMO519PH/RCrmCHERRYTAGCATAPHVIII
pMO520PH/RCrmCHERRY-3FLAGTAGCATAPHVIII
  • 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.