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Table 2 Plasmids used

From: Steroid biotransformations in biphasic systems with Yarrowia lipolytica expressing human liver cytochrome P450 genes

Plasmid Description Marker gene (selection in yeast) Reference
p64ICL1 Parental vector for multicopy integration in Y. lipolytica ura3d4 (mc)a [51]
p65ICL1 Parental vector for single copy integration in Y. lipolytica ura3d1 (sc) a [51]
p67RYL Source for Y. lipolytica cytochrome P450 reductase (YlCPRwt) ura3d4 (mc)a [22, 49]
pNMTS-CYP-OR-spe Source for codon optimized human cytochrome P450 3A4 (3A4syn)   Weis et al., unpublished (IMBT, TU Graz)
pBdpTrcRed-3A4(192v)wtA305S Source for codon optimized human cytochrome P450 reductase (hCPRsyn)   Weis et al., unpublished (IMBT, TU Graz)
ID7262313 Source for cDNA wild-type human cytochrome P450 reductase (hCPRwt)   BioCat, Germany
Cloning vectors:
pJet1.2 E. coli cloning vector   Fermentas, Germany
p64D-linker Cloning and basic integrative multicopy vector ura3d4 (mc) a This work
p65D-linker Cloning and basic integrative single copy vector, negative control ura3d1 (sc) a This work
Integrative vectors for CPR and CYP expression in Y. lipolytica:
p64D-hCPRwt-2D6syn Multicopy coexpression of human wild-type CPR (hCPRwt), codon optimized CPR (hCPRsyn) or Y.lipolytica-own CPR (YlCPR) in combination with codon optimized CYP2D6 or codon optimized CYP3A4, respectively. ura3d4 (mc) a This work
p64D-hCPRsyn-2D6syn    
p64D-hCPRwt-3A4syn    
p64D-YlCPR-2D6syn    
p64D-YlCPR-3A4syn    
p65D-hCPRwt-2D6syn Single copy coexpression of human wild-type CPR (hCPRwt) or Y. lipolytica-own CPR (YlCPR) in combination with codon optimized CYP2D6 or codon optimized CYP3A4, respectively. ura3d4 (mc) a This work
p65D-hCPRwt-3A4syn    
p65D-YlCPR-2D6syn    
p65D-YlCPR-3A4syn    
  1. aURA3 marker genes for single copy (sc, ura3d1) or multicopy (mc, ura3d4) selection with different promoter lengths.