Table 1.

Plasmids and primers used in this study

Plasmid Plasmid backbone Sequence, 5′-3′ 
Expression in Dictyostelium cells 
GFP-Leep2A pDM317 F: 5′-GGA​AGA​TCT​ATG​AGC​TCA​ACA​GCG​AAT​AAT​TCA​C-3′ 
R: 5′-GCT​CTA​GAA​ATT​GGT​GAA​GGT​ATA​CCA​ATT​GTT​G-3′ 
Flag-Leep2A pDM320 F: 5′-AGA​TGA​TGA​TGA​TAA​AAG​ATC​TAT​GAG​CTC​AAC​AGC​GAA​TAA​TTC​AC-3′ 
R: 5′-TAA​ATA​ATT​TAT​TTA​TTT​AAC​TAG​TTT​AAA​TTG​GTG​AAG​GTA​TAC​C-3′ 
RFP-Leep2A pDM449 F: 5′-GGA​AGA​TCT​ATG​AGC​TCA​ACA​GCG​AAT​AAT​TCA​C-3′ 
R: 5′-GCT​CTA​GAA​ATT​GGT​GAA​GGT​ATA​CCA​ATT​GTT​G-3′ 
GFP-Leep2AΔGAP pDM317 F: 5′-CTA​TCT​AGA​ATG​AGC​TCA​ACA​GCG​AAT​AAT​TCA​C-3′ 
F: 5′-CTA​GTC​GAC​ACC​ACC​TTG​ACT​ATT​GTA​TTG​TTT​G-3′ 
GFP-Leep2A1-1344 (T1) pDM317 F: 5′-GGA​AGA​TCT​ATG​AGC​TCA​ACA​GCG​AAT​AAT​TCA​C-3′ 
R: 5′-GTC​GAC​TTA​TTC​CAA​TTG​ACG​TAG​AGT​GTT​GGT​ATA​G-3′ 
GFP-Leep2A1-1100 (T2) pDM317 F: 5′-GGA​AGA​TCT​ATG​AGC​TCA​ACA​GCG​AAT​AAT​TCA​C-3′ 
R: 5′-GTC​GAC​TTA​AAT​GAA​TTC​TGC​ACC​TTG​TTT​TGA​TG-3′ 
GFP-Leep2A1-446 (T3) pDM317 F: 5′-GGA​AGA​TCT​ATG​AGC​TCA​ACA​GCG​AAT​AAT​TCA​C-3′ 
R: 5′-ACG​CGT​CGA​CTG​ATT​CTG​CTT​CGA​ATA​CAT​GTA​AC-3′ 
GFP-Leep2A447-1100 (T4) pDM317 F: 5′-CTA​TCT​AGA​ATG​CAA​AAT​TTT​GAA​AAA​GAG​AGA​GAA​GC-3′ 
R: 5′-GTC​GAC​TTA​AAT​GAA​TTC​TGC​ACC​TTG​TTT​TGA​TG-3′ 
GFP-Leep2A1101-1344 (T5) pDM317 F: 5′-TCT​AGA​AAA​GGT​ACC​AAA​AGA​TCT​ATG​AGT​TCA​ATG​ATT​GAT​GAA​GAA​G-3′ 
R: 5′-TTA​AAT​AAT​TTA​TTT​ATT​TAA​CTA​GTT​TCC​AAT​TGA​CGT​AGA​GTG​TTG​G-3′ 
GFP-Leep2A1345-1640 (T6) pDM317 F: 5′-GAA​GAT​CTA​TGT​CAA​CAA​ATA​AAT​TAA​GTC​GTG​CTC-3′ 
R: 5′-GCT​CTA​GAA​ATT​GGT​GAA​GGT​ATA​CCA​ATT​GTT​G-3′ 
GFP-Leep2AN1474K pDM317 F: 5′-TCA​TGT​GGG​TAA​AGA​TAT​TGT​AAA​CAT​CAT​TTG-3′ 
R: 5′-ACA​ATA​TCT​TTA​CCC​ACA​TGA​CGT​TTC​TTA​TGA​ATT​TG-3′ 
GFP-Leep2B pDM317 F: 5′-CTA​TCT​AGA​ATG​ATT​ACA​AAC​TCA​TTT​ACT​AAT​AAT​TTT​C-3′ 
F: 5′-CTA​GTC​GAC​TTT​ATG​AGA​TTG​ATC​TTC​AGA​TTG-3′ 
GFP-Leep2B pDM317-HygR F: 5′-CTA​TCT​AGA​ATG​ATT​ACA​AAC​TCA​TTT​ACT​AAT​AAT​TTT​C-3′ 
F: 5′-CTA​GTC​GAC​TTT​ATG​AGA​TTG​ATC​TTC​AGA​TTG-3′ 
RFP-Leep2B pDM449 F: 5′-CTA​TCT​AGA​ATG​ATT​ACA​AAC​TCA​TTT​ACT​AAT​AAT​TTT​C-3′ 
F: 5′-CTA​GTC​GAC​TTT​ATG​AGA​TTG​ATC​TTC​AGA​TTG-3′ 
RFP-Leep2BΔGAP pDM449 F: 5′-CCG​GAT​CCA​TGA​TTA​CAA​ACT​CAT​TTA​CTA​ATA-3′ 
R: 5′-CTA​TCT​AGA​ATC​GTC​TTC​GGT​TGA​GTC​ACC​CTT-3′ 
RFP-Leep2B1-1264 (T1) pDM449 F: 5′-CTA​TCT​AGA​ATG​ATT​ACA​AAC​TCA​TTT​ACT​AAT​AAT​TTT​C-3′ 
R:5′-GTCGACTTACTCCAATGGTGTCAATTTATTTCTATTC-3′ 
RFP-Leep2B1-1010 (T2) pDM449 F: 5′-CTA​TCT​AGA​ATG​ATT​ACA​AAC​TCA​TTT​ACT​AAT​AAT​TTT​C-3′ 
R:5′-GTCGACTTAGGTATTGGTTGGTGTAGCATTATATGG-3′ 
RFP-Leep2B1-400 (T3) pDM449 F: 5′-CTA​TCT​AGA​ATG​ATT​ACA​AAC​TCA​TTT​ACT​AAT​AAT​TTT​C-3′ 
R: 5′-CTA​GTC​GAC​ACT​ATT​GGA​ATG​AAT​ACC​AAG​TAT​AC-3′ 
RFP-Leep2B401-1010 (T4) pDM449 F: 5′-TCT​AGA​AAA​GGT​ACC​AAA​AGA​TCT​ATG​GAA​ATG​TTG​GCA​CGT​AAT​TTA​TG-3′ 
R: 5′-TAA​ATA​ATT​TAT​TTA​TTT​AAC​TAG​TTT​AGG​TAT​TGG​TTG​GTG​TAG​CAT​TAT​ATG-3′ 
RFP-Leep2B1011-1264 (T5) pDM449 F: 5′-TCT​AGA​AAA​GGT​ACC​AAA​AGA​TCT​ATG​TCA​TCA​AAA​GTA​ACC​GAA​TCT​G-3′ 
R: 5′-TTA​AAT​AAT​TTA​TTT​ATT​TAA​CTA​GTT​TAC​TCC​AAT​GGT​GTC​AAT​TTA​TTT​CTA​TTC-3′ 
RFP-Leep2B1265-1619 (T6) pDM449 F: 5′-TGC​TCT​AGA​ATG​AAT​AGT​GTT​CAA​TTT​TTC​CAA​TCT​TTA​AAT​ATG-3′ 
R: 5′-CTA​GTC​GAC​TTT​ATG​AGA​TTG​ATC​TTC​AGA​TTG-3′ 
Leep2B pDM344 F: 5′-CCG​GAT​CCT​ACC​CAT​ACG​ATG​TTC​CAG​ATT​ACG​CTA​TGA​TTA​CAA​ACT​CAT​TTA​CTA​ATA​ATT​TTC-3′ 
R: 5′-CTA​TCT​AGA​TTT​ATG​AGA​TTG​ATC​TTC​AGA​TTG-3′ 
Leep2BΔGAP pDM344 F: 5′-CCG​GAT​CCT​ACC​CAT​ACG​ATG​TTC​CAG​ATT​ACG​CTA​TGA​TTA​CAA​ACT​CAT​TTA​CTA​ATA​ATT​TTC-3′ 
R: 5′-CTA​TCT​AGA​ATC​GTC​TTC​GGT​TGA​GTC​ACC​CTT-3′ 
GFP-RasB pDM317 F: 5′-GGA​AGA​TCT​ATG​TCA​GTT​TCA​AAT​GAA​TAT​AAA​TTA​G-3′ 
R: 5′-GGA​CTA​GTC​TAA​AGG​ATT​AAA​CAA​TCA​CCA​CC-3′ 
GFP-RasD pDM317 F: 5′-GGG​GAT​CCA​TGA​CAG​AAT​ATA​AAT​TAG​TTA​TTG​TAG​GTG​G-3′ 
R: 5′-GGA​ACT​AGT​TTA​TAA​AAT​TAA​ACA​TTG​TTT​TTT​CTT​TTT​TTG​AGC-3′ 
GFP-RasG pDM317 F: 5′-GGG​GAT​CCA​TGA​CAG​AAT​ACA​AAT​TAG​TTA​TTG​TTG​G-3′ 
R: 5′-GGA​ACT​AGT​TTA​TAA​AAG​AGT​ACA​AGC​TTT​TAA​TGG​TC-3′ 
GFP-RasBG15V pDM371 F: 5′-GGA​AGA​TCT​ATG​TCA​GTT​TCA​AAT​GAA​TAT​AAA​TTA​G-3′ 
R: 5′-GGA​CTA​GTC​TAA​AGG​ATT​AAA​CAA​TCA​CCA​CC-3′ 
F: 5′-TTG​TTA​TGG​GTG​GTG​TTG​GTG​TTG​GTA​AGA​GTG-3′ 
R: 5′-TCT​TAC​CAA​CAC​CAA​CAC​CAC​CCA​TAA​CAA​CTA​ATT​TAT​ATT​C-3′ 
GFP-RasDQ61L pDM371 F: 5′-GGG​GAT​CCA​TGA​CAG​AAT​ATA​AAT​TAG​TTA​TTG​TAG​GTG​G-3′ 
R: 5′-GGA​ACT​AGT​TTA​TAA​AAT​TAA​ACA​TTG​TTT​TTT​CTT​TTT​TTG​AGC-3′ 
F: 5′-ACT​GCA​GGT​TTA​GAG​GAA​TAT​AGT​GCA​ATG-3′ 
R: 5′-TAT​TCC​TCT​AAA​CCT​GCA​GTA​TCT​AAA​ATA​TCT​AAT​AAA​C-3′ 
GFP-RasGG12V pDM371 F: 5′-GGG​GAT​CCA​TGA​CAG​AAT​ACA​AAT​TAG​TTA​TTG​TTG​G-3′ 
R: 5′-GGA​ACT​AGT​TTA​TAA​AAG​AGT​ACA​AGC​TTT​TAA​TGG​TC-3′ 
F: 5′-TGT​TGG​TGG​TGT​TGG​TGT​CGG​TAA​AAG​TGC​CTT​AAC-3′ 
R: 5′-TTT​ACC​GAC​ACC​AAC​ACC​ACC​AAC​AAT​AAC​TAA​TTT​GTA​TTC-3′ 
Gene distruption 
leep2A KO pBluescript-BSR ARM1-F: 5′-CGG​GGT​ACC​CAA​TCC​TCA​CAA​TCA​CCA​GTT​GCG-3′ 
ARM1-R: 5′-CCC​AAG​CTT​GTA​CAT​TTT​CAA​TAC​CTA​ACA​TTT​TCT-3′ 
ARM2-F: 5′-CGG​GGA​TCC​GGT​TGA​GGC​CAT​CTT​TCA​CGA​TAT​C-3′ 
ARM2-R: 5′-CCG​GCG​GCC​GCA​ATT​GTT​GAT​GGT​GGA​GTT​GTT​GAA​G-3′ 
P1: 5′-GGA​AGA​TCT​ATG​AGC​TCA​ACA​GCG​AAT​AAT​TCA​C-3′ 
P2: 5′-GTA​GCG​ACA​GAG​AAG​ATT​ACA​ATG​C-3′ 
P3: 5′-TCG​GGT​ATA​TTT​GAG​TGG​AAT​GAG-3′ 
P4: 5′-GCT​CTA​GAA​ATT​GGT​GAA​GGT​ATA​CCA​ATT​GTT​G-3′ 
leep2B KO pBluescript-BSR ARM1-F: 5′-CCG​GTA​CCA​TGA​TTA​CAA​ACT​CAT​TTA​CTA​A-3′ 
ARM1-R: 5′-CAA​GCT​TTG​GTG​AAT​TAT​TAT​TAC​TTG​ATT​C-3′ 
ARM2-F: 5′-CCG​GAT​CCA​TTA​GTA​AAC​ATA​TAC​TTG​CC-3′ 
ARM2-R: 5′-ATA​AGG​CGG​CCG​CTT​TAT​GAG​ATT​GAT​CTT​CAG​ATT​G-3′ 
P1: 5′-ATG​TTT​CTT​GAC​TGG​GCC​TCA​TCC-3′ 
P2: 5′-GTA​GCG​ACA​GAG​AAG​ATT​ACA​ATG​C-3′ 
P3: 5′-TCG​GGT​ATA​TTT​GAG​TGG​AAT​GAG-3′ 
P4: 5′-CCC​AGT​CAT​TGC​ACC​AGC​CAA​GCC-3′ 
rasB KO pBluescript-BSR ARM1-F: 5′-CGG​GGT​ACC​CAA​TTC​CAA​TAG​TAA​AAA​GTC-3′ 
ARM1-R: 5′-CTG​GTC​GAC​GGA​TAG​TAA​GTG​CAC​TCT​TAC​TG-3′ 
ARM2-F: 5′-CGG​GAT​CCG​TCA​AGA​TGA​TTA​CAG​TGC​TA-3′ 
ARM2-R: 5′-CCG​GCG​GCC​GCC​TTT​GAT​CTG​CCT​GGC​TCT​TTG-3′ 
P1: 5′-AAA​AAG​TAT​GAG​ATT​TTT​AAA​GTT​GG-3′ 
P2: 5′-GTA​GCG​ACA​GAG​AAG​ATT​ACA​ATG​C-3′ 
P4: 5′-ATC​ATC​TTG​ACC​TGC​AGT​ATC-3′ 
rasD KO pDM1801 ARM1-F: 5′-ATT​GCC​GGC​TTG​AAT​AGA​TAT​AAG​AGG​TCA​TAT​TC-3′ 
ARM1-R: 5′-CCC​AAG​CTT​ACC​ACC​TAC​AAT​AAC​TAA​TTT​ATA​TTC​TGT​C-3′ 
ARM2-F: 5′-GGA​AGA​TCT​ATC​AAT​CAA​GTG​GCA​AAG​CTC-3′ 
ARM2-R: 5′-GGA​ACT​AGT​ACT​ACA​AGC​ACT​TTT​CCA​GAT​ACC-3′ 
P1: 5′-ATC​TAT​TTG​TCA​GTG​AAT​TTA​TAC​C-3′ 
P2: 5′-TTT​ATC​GGC​ACT​TTG​CAT​CGG-3′ 
P3: 5′-AAA​CTT​CTC​GAC​AGA​CGT​CG-3′ 
P4: 5′-TAA​ATC​CTG​CCA​ATG​TAA​TTT​CAG​TAC-3′ 
P5: 5′-TAT​GAT​CCA​ACA​ATT​GAA​GAT​AGT​TAT​CG-3′ 
P6: 5′-AAT​TCT​ACT​TTT​AGC​AGA​TGA​CTC​C-3′ 
rasG KO pLPBLP ARM1-F: 5′-GAA​TTC​ATC​TAC​ACA​TGC​GAT​TAA​CTG​CCA​C-3′ 
ARM1-R: 5′-CAT​TGA​TGA​ATA​CGA​TCC​AAC​TAT​CG-3′ 
ARM2-F: 5′-GCG​GCC​GCG​TGT​CTA​CTC​TAT​CAC​TTC​AAG-3′ 
ARM2-R: 5′-GTA​AAT​TAG​AAA​TAA​TAA​TTC​AAC​CAC​CGC​GG-3′ 
P1: 5′-AAA​TCT​ACA​CAT​GCG​ATT​AAC-3′ 
P2: 5′-GTA​GCG​ACA​GAG​AAG​ATT​ACA​ATG​C-3′ 
P3: 5′-TCG​GGT​ATA​TTT​GAG​TGG​AAT​GAG-3′ 
P4: 5′-TTT​TAA​TTG​ATC​GGA​TTG​GA-3′ 
Expression in bacteria 
GST-RapA pGEX-6p-1 F: 5′-GCG​GAT​CCA​TGC​CTC​TTA​GAG​AAT​TCA​AAA​TCG-3′ 
R: 5′-ACG​CGT​CGA​CTT​ACA​ATA​AAG​CAC​ATT​TTG​ATT​TAG​C-3′ 
GST-RapB pGEX-6p-1 F: 5′-GCG​GAT​CCA​TGG​GTA​AGG​GAA​ATG​GTA​AAT​C-3′ 
R: 5′-ACG​CGT​CGA​CTT​ACA​TAA​TGA​TAC​ATT​TTT​CTT​TT-3′ 
GST-RapC pGEX-6p-1 F: 5′-GCG​GAT​CCA​TGC​AAA​CCT​ATA​AAG​TAG​TTG​TTT​TG-3′ 
R: 5′-ACG​CGT​CGA​CTT​ACA​TGA​TTA​AAC​ATT​TTC​CTT​TTG-3′ 
GST-RanA pGEX-6p-1 F: 5′-GCG​GAT​CCA​TGG​CAG​AAA​AAG​AAC​AAA​TTA​AAT​TAG-3′ 
R: 5′-ACG​CGT​CGA​CTT​ACA​AGT​CAT​CAT​TGT​CTT​CTG​G-3′ 
GST-RasB pGEX-6p-1 F: 5′-GGG​GAT​CCA​TGT​CAG​TTT​CAA​ATG​AAT​ATA​AAT​TAG-3′ 
R: 5′-CGG​AAT​TCC​TAA​AGG​ATT​AAA​CAA​TCA​CCA​C-3′ 
GST-RasC pGEX-6p-1 F: 5′-GGG​GAT​CCA​TGT​CAA​AAT​TAT​TAA​AAT​TAG​TTA​TCG​TTG​G-3′ 
R: 5′-CGG​AAT​TCT​TAC​AAT​ATA​ATA​CAT​CCC​CTT​TTC​TTT​GG-3′ 
GST-RasD pGEX-6p-1 F: 5′-GGG​GAT​CCA​TGA​CAG​AAT​ATA​AAT​TAG​TTA​TTG​TAG​GTG​G-3′ 
R: 5′-CGG​AAT​TCT​TAT​AAA​ATT​AAA​CAT​TGT​TTT​TTC​TTT​TTT​TGA​GC-3′ 
GST-RasG pGEX-6p-1 F: 5′-GGG​GAT​CCA​TGA​CAG​AAT​ACA​AAT​TAG​TTA​TTG​TTG​G-3′ 
R: 5′-CGG​AAT​TCT​TAT​AAA​AGA​GTA​CAA​GCT​TTT​AAT​GGT​C-3′ 
GST-RasS pGEX-6p-1 F: 5′-GGG​GAT​CCA​TGT​TTA​ATT​TTA​AAT​TAG​TAT​TAG​TTG​G-3′ 
R: 5′-CGG​AAT​TCT​TAT​AAT​AAA​TTA​CAA​GAT​TTC​TTT​TTT​TTA​ATT​GGT​TG-3′ 
GST-RasU pGEX-6p-1 F: 5′-CGG​GAT​CCA​TGT​CAG​CCT​TTA​TAT​ATA​ATA​ACA​G-3′ 
R: 5′-GAG​TCG​ACT​TAT​ATC​ATT​TTA​CAA​ATA​GAA​TGA​AAT​G-3′ 
GST-RasV pGEX-6p-1 F: 5′-CGG​GAT​CCA​TGT​CAA​TTA​AAA​TTA​AAA​ATT​TTA​AAA​AAA​ATA-3′ 
R: 5′-GAG​TCG​ACT​TAC​ATA​ACT​TTA​CAA​ATT​TCA​TTA​ATT​TTT​TTT​TTT​AAT​G-3′ 
GST-RasW pGEX-6p-1 F: 5′-CGG​GAT​CCA​TGA​CTT​CCT​ATA​AAA​ATA​ATA​ATG-3′ 
R: 5′-GAG​TCG​ACT​TAC​ATC​ATT​TTA​CAA​ATT​GAA​GTT​TTA​G-3′ 
GST-RasX pGEX-6p-1 F: 5′-CGG​GAT​CCA​TGT​CAG​GAT​ATA​AAA​ATA​ATA​ATT​TAG-3′ 
R: 5′-GAG​TCG​ACT​TAC​ATC​ATT​TTA​CAA​ATT​GAA​TTT​TTA​G-3′ 
GST-RasY pGEX-6p-1 F: 5′-CGG​GAT​CCA​TGA​CAA​CAA​ATA​AAA​GTA​ATG​GTA​ATT​TAG-3′ 
R: 5′-GAG​TCG​ACT​TAC​ATC​ATT​TTA​CAA​ATT​GAT​ATT​TTG​GC-3′ 
GST-RasZ pGEX-6p-1 F: 5′-CGG​GAT​CCA​TGG​CAT​CAT​ATA​AAA​ATA​ATA​ATT​TAG-3′ 
R: 5′-GAG​TCG​ACT​TAC​ATC​ATT​TTA​CAA​ATT​GAG​GTT​TTA​G-3′ 
Yeast two-hybrid 
Leep2A1359-1560 pGADT7 F: 5′-CGC​CAT​ATG​ACT​CCT​GGT​CGT​GAA​ATC​TTA​AAG-3′ 
R: 5′-CGG​GAT​CCT​TAT​GGA​GTA​TTA​TGA​CGA​ACA​TAA​CG-3′ 
RasBQ64L ΔCAAX pGBKT7 F: 5′-CGG​AAT​TCA​TGT​CAG​TTT​CAA​ATG​AAT​ATA​AAT​TAG-3′ 
R: 5′-CGG​GAT​CCT​TAA​TCA​CCA​CCT​TTA​AAC​TTT​TTA​AGG-3′ 
RasCQ62L ΔCAAX pGBKT7 F: 5′-CGG​AAT​TCA​TGT​CAA​AAT​TAT​TAA​AAT​TAG​TTA​TC-3′ 
R: 5′-CGG​GAT​CCT​TAT​CCC​CTT​TTC​TTT​GGT​GGG-3′ 
RasDQ61L ΔCAAX pGBKT7 F: 5′-CGG​AAT​TCA​TGA​CAG​AAT​ATA​AAT​TAG​TTA​TTG-3′ 
R: 5′-CGG​GAT​CCT​TAT​TGT​TTT​TTC​TTT​TTT​TGA​GC-3′ 
RasGQ61L ΔCAAX pGBKT7 F: 5′-CGG​AAT​TCA​TGA​CAG​AAT​ACA​AAT​TAG​TTA​TTG-3′ 
R: 5′-CGG​GAT​CCT​TAA​GCT​TTT​AAT​GGT​CTC​TTC-3′ 
RasSQ61L ΔCAAX pGBKT7 F: 5′-CGG​AAT​TCA​TGT​TTA​ATT​TTA​AAT​TAG​TAT​TAG​TTG-3′ 
R: 5′-CGG​GAT​CCT​TAA​GAT​TTC​TTT​TTT​TTA​ATT​GG-3′ 
RasUQ72L ΔCAAX pGBKT7 F: 5′-GGC​ATA​TGT​CAG​CCT​TTA​TAT​ATA​ATA​ACA​G-3′ 
R: 5′-CGG​GAT​CCT​TAA​ATA​GAA​TGA​AAT​GTA​TTA​TTT​AAT​TTT​TG-3′ 
RasVQ91L ΔCAAX pGBKT7 F: 5′-CGG​AAT​TCA​TGT​CAA​TTA​AAA​TTA​AAA​ATT​TTA-3′ 
R: 5′-CGG​GAT​CCT​TAA​ATT​TCA​TTA​ATT​TTT​TTT​TTT​AAT​GG-3′ 
RasWQ67L ΔCAAX pGBKT7 F: 5′-CGG​AAT​TCA​TGA​CTT​CCT​ATA​AAA​ATA​ATA​ATG-3′ 
R: 5′-CGG​GAT​CCT​TAA​ATT​GAA​GTT​TTA​GTT​TG-3′ 
RasXQ67L ΔCAAX pGBKT7 F: 5′-CGG​AAT​TCA​TGT​CAG​GAT​ATA​AAA​ATA​ATA​AT-3′ 
R: 5′-CGG​GAT​CCT​TAA​ATT​GAA​TTT​TTA​GTA​TTA​TTT​AC-3′ 
RasYQ68L ΔCAAX pGBKT7 F: 5′-CGG​AAT​TCA​TGA​CAA​CAA​ATA​AAA​GTA​ATG​G-3′ 
R: 5′-CGG​GAT​CCT​TAA​ATT​GAT​ATT​TTG​GCA​CCA​TTT​ACT​TTT​TG-3′ 
RasZQ67L ΔCAAX pGBKT7 F: 5′-CGG​AAT​TCA​TGG​CAT​CAT​ATA​AAA​ATA​ATA​AT-3′ 
R: 5′-CGG​GAT​CCT​TAA​ATT​GAG​GTT​TTA​GTA​TTA​TTT​AC-3′ 
RanAQ65L pGBKT7 F: 5′-GGC​ATA​TGG​CAG​AAA​AAG​AAC​AAA​TTA​AAT​TAG-3′ 
R: 5′-GCG​GAT​CCT​TAC​AAG​TCA​TCA​TTG​TCT​TCT​GG-3′ 
RapAQ65E ΔCAAX pGBKT7 F: 5′-GGC​ATA​TGC​CTC​TTA​GAG​AAT​TCA​AAA​TCG-3′ 
R: 5′-CGG​GAT​CCT​TAT​TTT​GAT​TTA​GCT​TTG​CTT​GG-3′ 
Plasmid Plasmid backbone Sequence, 5′-3′ 
Expression in Dictyostelium cells 
GFP-Leep2A pDM317 F: 5′-GGA​AGA​TCT​ATG​AGC​TCA​ACA​GCG​AAT​AAT​TCA​C-3′ 
R: 5′-GCT​CTA​GAA​ATT​GGT​GAA​GGT​ATA​CCA​ATT​GTT​G-3′ 
Flag-Leep2A pDM320 F: 5′-AGA​TGA​TGA​TGA​TAA​AAG​ATC​TAT​GAG​CTC​AAC​AGC​GAA​TAA​TTC​AC-3′ 
R: 5′-TAA​ATA​ATT​TAT​TTA​TTT​AAC​TAG​TTT​AAA​TTG​GTG​AAG​GTA​TAC​C-3′ 
RFP-Leep2A pDM449 F: 5′-GGA​AGA​TCT​ATG​AGC​TCA​ACA​GCG​AAT​AAT​TCA​C-3′ 
R: 5′-GCT​CTA​GAA​ATT​GGT​GAA​GGT​ATA​CCA​ATT​GTT​G-3′ 
GFP-Leep2AΔGAP pDM317 F: 5′-CTA​TCT​AGA​ATG​AGC​TCA​ACA​GCG​AAT​AAT​TCA​C-3′ 
F: 5′-CTA​GTC​GAC​ACC​ACC​TTG​ACT​ATT​GTA​TTG​TTT​G-3′ 
GFP-Leep2A1-1344 (T1) pDM317 F: 5′-GGA​AGA​TCT​ATG​AGC​TCA​ACA​GCG​AAT​AAT​TCA​C-3′ 
R: 5′-GTC​GAC​TTA​TTC​CAA​TTG​ACG​TAG​AGT​GTT​GGT​ATA​G-3′ 
GFP-Leep2A1-1100 (T2) pDM317 F: 5′-GGA​AGA​TCT​ATG​AGC​TCA​ACA​GCG​AAT​AAT​TCA​C-3′ 
R: 5′-GTC​GAC​TTA​AAT​GAA​TTC​TGC​ACC​TTG​TTT​TGA​TG-3′ 
GFP-Leep2A1-446 (T3) pDM317 F: 5′-GGA​AGA​TCT​ATG​AGC​TCA​ACA​GCG​AAT​AAT​TCA​C-3′ 
R: 5′-ACG​CGT​CGA​CTG​ATT​CTG​CTT​CGA​ATA​CAT​GTA​AC-3′ 
GFP-Leep2A447-1100 (T4) pDM317 F: 5′-CTA​TCT​AGA​ATG​CAA​AAT​TTT​GAA​AAA​GAG​AGA​GAA​GC-3′ 
R: 5′-GTC​GAC​TTA​AAT​GAA​TTC​TGC​ACC​TTG​TTT​TGA​TG-3′ 
GFP-Leep2A1101-1344 (T5) pDM317 F: 5′-TCT​AGA​AAA​GGT​ACC​AAA​AGA​TCT​ATG​AGT​TCA​ATG​ATT​GAT​GAA​GAA​G-3′ 
R: 5′-TTA​AAT​AAT​TTA​TTT​ATT​TAA​CTA​GTT​TCC​AAT​TGA​CGT​AGA​GTG​TTG​G-3′ 
GFP-Leep2A1345-1640 (T6) pDM317 F: 5′-GAA​GAT​CTA​TGT​CAA​CAA​ATA​AAT​TAA​GTC​GTG​CTC-3′ 
R: 5′-GCT​CTA​GAA​ATT​GGT​GAA​GGT​ATA​CCA​ATT​GTT​G-3′ 
GFP-Leep2AN1474K pDM317 F: 5′-TCA​TGT​GGG​TAA​AGA​TAT​TGT​AAA​CAT​CAT​TTG-3′ 
R: 5′-ACA​ATA​TCT​TTA​CCC​ACA​TGA​CGT​TTC​TTA​TGA​ATT​TG-3′ 
GFP-Leep2B pDM317 F: 5′-CTA​TCT​AGA​ATG​ATT​ACA​AAC​TCA​TTT​ACT​AAT​AAT​TTT​C-3′ 
F: 5′-CTA​GTC​GAC​TTT​ATG​AGA​TTG​ATC​TTC​AGA​TTG-3′ 
GFP-Leep2B pDM317-HygR F: 5′-CTA​TCT​AGA​ATG​ATT​ACA​AAC​TCA​TTT​ACT​AAT​AAT​TTT​C-3′ 
F: 5′-CTA​GTC​GAC​TTT​ATG​AGA​TTG​ATC​TTC​AGA​TTG-3′ 
RFP-Leep2B pDM449 F: 5′-CTA​TCT​AGA​ATG​ATT​ACA​AAC​TCA​TTT​ACT​AAT​AAT​TTT​C-3′ 
F: 5′-CTA​GTC​GAC​TTT​ATG​AGA​TTG​ATC​TTC​AGA​TTG-3′ 
RFP-Leep2BΔGAP pDM449 F: 5′-CCG​GAT​CCA​TGA​TTA​CAA​ACT​CAT​TTA​CTA​ATA-3′ 
R: 5′-CTA​TCT​AGA​ATC​GTC​TTC​GGT​TGA​GTC​ACC​CTT-3′ 
RFP-Leep2B1-1264 (T1) pDM449 F: 5′-CTA​TCT​AGA​ATG​ATT​ACA​AAC​TCA​TTT​ACT​AAT​AAT​TTT​C-3′ 
R:5′-GTCGACTTACTCCAATGGTGTCAATTTATTTCTATTC-3′ 
RFP-Leep2B1-1010 (T2) pDM449 F: 5′-CTA​TCT​AGA​ATG​ATT​ACA​AAC​TCA​TTT​ACT​AAT​AAT​TTT​C-3′ 
R:5′-GTCGACTTAGGTATTGGTTGGTGTAGCATTATATGG-3′ 
RFP-Leep2B1-400 (T3) pDM449 F: 5′-CTA​TCT​AGA​ATG​ATT​ACA​AAC​TCA​TTT​ACT​AAT​AAT​TTT​C-3′ 
R: 5′-CTA​GTC​GAC​ACT​ATT​GGA​ATG​AAT​ACC​AAG​TAT​AC-3′ 
RFP-Leep2B401-1010 (T4) pDM449 F: 5′-TCT​AGA​AAA​GGT​ACC​AAA​AGA​TCT​ATG​GAA​ATG​TTG​GCA​CGT​AAT​TTA​TG-3′ 
R: 5′-TAA​ATA​ATT​TAT​TTA​TTT​AAC​TAG​TTT​AGG​TAT​TGG​TTG​GTG​TAG​CAT​TAT​ATG-3′ 
RFP-Leep2B1011-1264 (T5) pDM449 F: 5′-TCT​AGA​AAA​GGT​ACC​AAA​AGA​TCT​ATG​TCA​TCA​AAA​GTA​ACC​GAA​TCT​G-3′ 
R: 5′-TTA​AAT​AAT​TTA​TTT​ATT​TAA​CTA​GTT​TAC​TCC​AAT​GGT​GTC​AAT​TTA​TTT​CTA​TTC-3′ 
RFP-Leep2B1265-1619 (T6) pDM449 F: 5′-TGC​TCT​AGA​ATG​AAT​AGT​GTT​CAA​TTT​TTC​CAA​TCT​TTA​AAT​ATG-3′ 
R: 5′-CTA​GTC​GAC​TTT​ATG​AGA​TTG​ATC​TTC​AGA​TTG-3′ 
Leep2B pDM344 F: 5′-CCG​GAT​CCT​ACC​CAT​ACG​ATG​TTC​CAG​ATT​ACG​CTA​TGA​TTA​CAA​ACT​CAT​TTA​CTA​ATA​ATT​TTC-3′ 
R: 5′-CTA​TCT​AGA​TTT​ATG​AGA​TTG​ATC​TTC​AGA​TTG-3′ 
Leep2BΔGAP pDM344 F: 5′-CCG​GAT​CCT​ACC​CAT​ACG​ATG​TTC​CAG​ATT​ACG​CTA​TGA​TTA​CAA​ACT​CAT​TTA​CTA​ATA​ATT​TTC-3′ 
R: 5′-CTA​TCT​AGA​ATC​GTC​TTC​GGT​TGA​GTC​ACC​CTT-3′ 
GFP-RasB pDM317 F: 5′-GGA​AGA​TCT​ATG​TCA​GTT​TCA​AAT​GAA​TAT​AAA​TTA​G-3′ 
R: 5′-GGA​CTA​GTC​TAA​AGG​ATT​AAA​CAA​TCA​CCA​CC-3′ 
GFP-RasD pDM317 F: 5′-GGG​GAT​CCA​TGA​CAG​AAT​ATA​AAT​TAG​TTA​TTG​TAG​GTG​G-3′ 
R: 5′-GGA​ACT​AGT​TTA​TAA​AAT​TAA​ACA​TTG​TTT​TTT​CTT​TTT​TTG​AGC-3′ 
GFP-RasG pDM317 F: 5′-GGG​GAT​CCA​TGA​CAG​AAT​ACA​AAT​TAG​TTA​TTG​TTG​G-3′ 
R: 5′-GGA​ACT​AGT​TTA​TAA​AAG​AGT​ACA​AGC​TTT​TAA​TGG​TC-3′ 
GFP-RasBG15V pDM371 F: 5′-GGA​AGA​TCT​ATG​TCA​GTT​TCA​AAT​GAA​TAT​AAA​TTA​G-3′ 
R: 5′-GGA​CTA​GTC​TAA​AGG​ATT​AAA​CAA​TCA​CCA​CC-3′ 
F: 5′-TTG​TTA​TGG​GTG​GTG​TTG​GTG​TTG​GTA​AGA​GTG-3′ 
R: 5′-TCT​TAC​CAA​CAC​CAA​CAC​CAC​CCA​TAA​CAA​CTA​ATT​TAT​ATT​C-3′ 
GFP-RasDQ61L pDM371 F: 5′-GGG​GAT​CCA​TGA​CAG​AAT​ATA​AAT​TAG​TTA​TTG​TAG​GTG​G-3′ 
R: 5′-GGA​ACT​AGT​TTA​TAA​AAT​TAA​ACA​TTG​TTT​TTT​CTT​TTT​TTG​AGC-3′ 
F: 5′-ACT​GCA​GGT​TTA​GAG​GAA​TAT​AGT​GCA​ATG-3′ 
R: 5′-TAT​TCC​TCT​AAA​CCT​GCA​GTA​TCT​AAA​ATA​TCT​AAT​AAA​C-3′ 
GFP-RasGG12V pDM371 F: 5′-GGG​GAT​CCA​TGA​CAG​AAT​ACA​AAT​TAG​TTA​TTG​TTG​G-3′ 
R: 5′-GGA​ACT​AGT​TTA​TAA​AAG​AGT​ACA​AGC​TTT​TAA​TGG​TC-3′ 
F: 5′-TGT​TGG​TGG​TGT​TGG​TGT​CGG​TAA​AAG​TGC​CTT​AAC-3′ 
R: 5′-TTT​ACC​GAC​ACC​AAC​ACC​ACC​AAC​AAT​AAC​TAA​TTT​GTA​TTC-3′ 
Gene distruption 
leep2A KO pBluescript-BSR ARM1-F: 5′-CGG​GGT​ACC​CAA​TCC​TCA​CAA​TCA​CCA​GTT​GCG-3′ 
ARM1-R: 5′-CCC​AAG​CTT​GTA​CAT​TTT​CAA​TAC​CTA​ACA​TTT​TCT-3′ 
ARM2-F: 5′-CGG​GGA​TCC​GGT​TGA​GGC​CAT​CTT​TCA​CGA​TAT​C-3′ 
ARM2-R: 5′-CCG​GCG​GCC​GCA​ATT​GTT​GAT​GGT​GGA​GTT​GTT​GAA​G-3′ 
P1: 5′-GGA​AGA​TCT​ATG​AGC​TCA​ACA​GCG​AAT​AAT​TCA​C-3′ 
P2: 5′-GTA​GCG​ACA​GAG​AAG​ATT​ACA​ATG​C-3′ 
P3: 5′-TCG​GGT​ATA​TTT​GAG​TGG​AAT​GAG-3′ 
P4: 5′-GCT​CTA​GAA​ATT​GGT​GAA​GGT​ATA​CCA​ATT​GTT​G-3′ 
leep2B KO pBluescript-BSR ARM1-F: 5′-CCG​GTA​CCA​TGA​TTA​CAA​ACT​CAT​TTA​CTA​A-3′ 
ARM1-R: 5′-CAA​GCT​TTG​GTG​AAT​TAT​TAT​TAC​TTG​ATT​C-3′ 
ARM2-F: 5′-CCG​GAT​CCA​TTA​GTA​AAC​ATA​TAC​TTG​CC-3′ 
ARM2-R: 5′-ATA​AGG​CGG​CCG​CTT​TAT​GAG​ATT​GAT​CTT​CAG​ATT​G-3′ 
P1: 5′-ATG​TTT​CTT​GAC​TGG​GCC​TCA​TCC-3′ 
P2: 5′-GTA​GCG​ACA​GAG​AAG​ATT​ACA​ATG​C-3′ 
P3: 5′-TCG​GGT​ATA​TTT​GAG​TGG​AAT​GAG-3′ 
P4: 5′-CCC​AGT​CAT​TGC​ACC​AGC​CAA​GCC-3′ 
rasB KO pBluescript-BSR ARM1-F: 5′-CGG​GGT​ACC​CAA​TTC​CAA​TAG​TAA​AAA​GTC-3′ 
ARM1-R: 5′-CTG​GTC​GAC​GGA​TAG​TAA​GTG​CAC​TCT​TAC​TG-3′ 
ARM2-F: 5′-CGG​GAT​CCG​TCA​AGA​TGA​TTA​CAG​TGC​TA-3′ 
ARM2-R: 5′-CCG​GCG​GCC​GCC​TTT​GAT​CTG​CCT​GGC​TCT​TTG-3′ 
P1: 5′-AAA​AAG​TAT​GAG​ATT​TTT​AAA​GTT​GG-3′ 
P2: 5′-GTA​GCG​ACA​GAG​AAG​ATT​ACA​ATG​C-3′ 
P4: 5′-ATC​ATC​TTG​ACC​TGC​AGT​ATC-3′ 
rasD KO pDM1801 ARM1-F: 5′-ATT​GCC​GGC​TTG​AAT​AGA​TAT​AAG​AGG​TCA​TAT​TC-3′ 
ARM1-R: 5′-CCC​AAG​CTT​ACC​ACC​TAC​AAT​AAC​TAA​TTT​ATA​TTC​TGT​C-3′ 
ARM2-F: 5′-GGA​AGA​TCT​ATC​AAT​CAA​GTG​GCA​AAG​CTC-3′ 
ARM2-R: 5′-GGA​ACT​AGT​ACT​ACA​AGC​ACT​TTT​CCA​GAT​ACC-3′ 
P1: 5′-ATC​TAT​TTG​TCA​GTG​AAT​TTA​TAC​C-3′ 
P2: 5′-TTT​ATC​GGC​ACT​TTG​CAT​CGG-3′ 
P3: 5′-AAA​CTT​CTC​GAC​AGA​CGT​CG-3′ 
P4: 5′-TAA​ATC​CTG​CCA​ATG​TAA​TTT​CAG​TAC-3′ 
P5: 5′-TAT​GAT​CCA​ACA​ATT​GAA​GAT​AGT​TAT​CG-3′ 
P6: 5′-AAT​TCT​ACT​TTT​AGC​AGA​TGA​CTC​C-3′ 
rasG KO pLPBLP ARM1-F: 5′-GAA​TTC​ATC​TAC​ACA​TGC​GAT​TAA​CTG​CCA​C-3′ 
ARM1-R: 5′-CAT​TGA​TGA​ATA​CGA​TCC​AAC​TAT​CG-3′ 
ARM2-F: 5′-GCG​GCC​GCG​TGT​CTA​CTC​TAT​CAC​TTC​AAG-3′ 
ARM2-R: 5′-GTA​AAT​TAG​AAA​TAA​TAA​TTC​AAC​CAC​CGC​GG-3′ 
P1: 5′-AAA​TCT​ACA​CAT​GCG​ATT​AAC-3′ 
P2: 5′-GTA​GCG​ACA​GAG​AAG​ATT​ACA​ATG​C-3′ 
P3: 5′-TCG​GGT​ATA​TTT​GAG​TGG​AAT​GAG-3′ 
P4: 5′-TTT​TAA​TTG​ATC​GGA​TTG​GA-3′ 
Expression in bacteria 
GST-RapA pGEX-6p-1 F: 5′-GCG​GAT​CCA​TGC​CTC​TTA​GAG​AAT​TCA​AAA​TCG-3′ 
R: 5′-ACG​CGT​CGA​CTT​ACA​ATA​AAG​CAC​ATT​TTG​ATT​TAG​C-3′ 
GST-RapB pGEX-6p-1 F: 5′-GCG​GAT​CCA​TGG​GTA​AGG​GAA​ATG​GTA​AAT​C-3′ 
R: 5′-ACG​CGT​CGA​CTT​ACA​TAA​TGA​TAC​ATT​TTT​CTT​TT-3′ 
GST-RapC pGEX-6p-1 F: 5′-GCG​GAT​CCA​TGC​AAA​CCT​ATA​AAG​TAG​TTG​TTT​TG-3′ 
R: 5′-ACG​CGT​CGA​CTT​ACA​TGA​TTA​AAC​ATT​TTC​CTT​TTG-3′ 
GST-RanA pGEX-6p-1 F: 5′-GCG​GAT​CCA​TGG​CAG​AAA​AAG​AAC​AAA​TTA​AAT​TAG-3′ 
R: 5′-ACG​CGT​CGA​CTT​ACA​AGT​CAT​CAT​TGT​CTT​CTG​G-3′ 
GST-RasB pGEX-6p-1 F: 5′-GGG​GAT​CCA​TGT​CAG​TTT​CAA​ATG​AAT​ATA​AAT​TAG-3′ 
R: 5′-CGG​AAT​TCC​TAA​AGG​ATT​AAA​CAA​TCA​CCA​C-3′ 
GST-RasC pGEX-6p-1 F: 5′-GGG​GAT​CCA​TGT​CAA​AAT​TAT​TAA​AAT​TAG​TTA​TCG​TTG​G-3′ 
R: 5′-CGG​AAT​TCT​TAC​AAT​ATA​ATA​CAT​CCC​CTT​TTC​TTT​GG-3′ 
GST-RasD pGEX-6p-1 F: 5′-GGG​GAT​CCA​TGA​CAG​AAT​ATA​AAT​TAG​TTA​TTG​TAG​GTG​G-3′ 
R: 5′-CGG​AAT​TCT​TAT​AAA​ATT​AAA​CAT​TGT​TTT​TTC​TTT​TTT​TGA​GC-3′ 
GST-RasG pGEX-6p-1 F: 5′-GGG​GAT​CCA​TGA​CAG​AAT​ACA​AAT​TAG​TTA​TTG​TTG​G-3′ 
R: 5′-CGG​AAT​TCT​TAT​AAA​AGA​GTA​CAA​GCT​TTT​AAT​GGT​C-3′ 
GST-RasS pGEX-6p-1 F: 5′-GGG​GAT​CCA​TGT​TTA​ATT​TTA​AAT​TAG​TAT​TAG​TTG​G-3′ 
R: 5′-CGG​AAT​TCT​TAT​AAT​AAA​TTA​CAA​GAT​TTC​TTT​TTT​TTA​ATT​GGT​TG-3′ 
GST-RasU pGEX-6p-1 F: 5′-CGG​GAT​CCA​TGT​CAG​CCT​TTA​TAT​ATA​ATA​ACA​G-3′ 
R: 5′-GAG​TCG​ACT​TAT​ATC​ATT​TTA​CAA​ATA​GAA​TGA​AAT​G-3′ 
GST-RasV pGEX-6p-1 F: 5′-CGG​GAT​CCA​TGT​CAA​TTA​AAA​TTA​AAA​ATT​TTA​AAA​AAA​ATA-3′ 
R: 5′-GAG​TCG​ACT​TAC​ATA​ACT​TTA​CAA​ATT​TCA​TTA​ATT​TTT​TTT​TTT​AAT​G-3′ 
GST-RasW pGEX-6p-1 F: 5′-CGG​GAT​CCA​TGA​CTT​CCT​ATA​AAA​ATA​ATA​ATG-3′ 
R: 5′-GAG​TCG​ACT​TAC​ATC​ATT​TTA​CAA​ATT​GAA​GTT​TTA​G-3′ 
GST-RasX pGEX-6p-1 F: 5′-CGG​GAT​CCA​TGT​CAG​GAT​ATA​AAA​ATA​ATA​ATT​TAG-3′ 
R: 5′-GAG​TCG​ACT​TAC​ATC​ATT​TTA​CAA​ATT​GAA​TTT​TTA​G-3′ 
GST-RasY pGEX-6p-1 F: 5′-CGG​GAT​CCA​TGA​CAA​CAA​ATA​AAA​GTA​ATG​GTA​ATT​TAG-3′ 
R: 5′-GAG​TCG​ACT​TAC​ATC​ATT​TTA​CAA​ATT​GAT​ATT​TTG​GC-3′ 
GST-RasZ pGEX-6p-1 F: 5′-CGG​GAT​CCA​TGG​CAT​CAT​ATA​AAA​ATA​ATA​ATT​TAG-3′ 
R: 5′-GAG​TCG​ACT​TAC​ATC​ATT​TTA​CAA​ATT​GAG​GTT​TTA​G-3′ 
Yeast two-hybrid 
Leep2A1359-1560 pGADT7 F: 5′-CGC​CAT​ATG​ACT​CCT​GGT​CGT​GAA​ATC​TTA​AAG-3′ 
R: 5′-CGG​GAT​CCT​TAT​GGA​GTA​TTA​TGA​CGA​ACA​TAA​CG-3′ 
RasBQ64L ΔCAAX pGBKT7 F: 5′-CGG​AAT​TCA​TGT​CAG​TTT​CAA​ATG​AAT​ATA​AAT​TAG-3′ 
R: 5′-CGG​GAT​CCT​TAA​TCA​CCA​CCT​TTA​AAC​TTT​TTA​AGG-3′ 
RasCQ62L ΔCAAX pGBKT7 F: 5′-CGG​AAT​TCA​TGT​CAA​AAT​TAT​TAA​AAT​TAG​TTA​TC-3′ 
R: 5′-CGG​GAT​CCT​TAT​CCC​CTT​TTC​TTT​GGT​GGG-3′ 
RasDQ61L ΔCAAX pGBKT7 F: 5′-CGG​AAT​TCA​TGA​CAG​AAT​ATA​AAT​TAG​TTA​TTG-3′ 
R: 5′-CGG​GAT​CCT​TAT​TGT​TTT​TTC​TTT​TTT​TGA​GC-3′ 
RasGQ61L ΔCAAX pGBKT7 F: 5′-CGG​AAT​TCA​TGA​CAG​AAT​ACA​AAT​TAG​TTA​TTG-3′ 
R: 5′-CGG​GAT​CCT​TAA​GCT​TTT​AAT​GGT​CTC​TTC-3′ 
RasSQ61L ΔCAAX pGBKT7 F: 5′-CGG​AAT​TCA​TGT​TTA​ATT​TTA​AAT​TAG​TAT​TAG​TTG-3′ 
R: 5′-CGG​GAT​CCT​TAA​GAT​TTC​TTT​TTT​TTA​ATT​GG-3′ 
RasUQ72L ΔCAAX pGBKT7 F: 5′-GGC​ATA​TGT​CAG​CCT​TTA​TAT​ATA​ATA​ACA​G-3′ 
R: 5′-CGG​GAT​CCT​TAA​ATA​GAA​TGA​AAT​GTA​TTA​TTT​AAT​TTT​TG-3′ 
RasVQ91L ΔCAAX pGBKT7 F: 5′-CGG​AAT​TCA​TGT​CAA​TTA​AAA​TTA​AAA​ATT​TTA-3′ 
R: 5′-CGG​GAT​CCT​TAA​ATT​TCA​TTA​ATT​TTT​TTT​TTT​AAT​GG-3′ 
RasWQ67L ΔCAAX pGBKT7 F: 5′-CGG​AAT​TCA​TGA​CTT​CCT​ATA​AAA​ATA​ATA​ATG-3′ 
R: 5′-CGG​GAT​CCT​TAA​ATT​GAA​GTT​TTA​GTT​TG-3′ 
RasXQ67L ΔCAAX pGBKT7 F: 5′-CGG​AAT​TCA​TGT​CAG​GAT​ATA​AAA​ATA​ATA​AT-3′ 
R: 5′-CGG​GAT​CCT​TAA​ATT​GAA​TTT​TTA​GTA​TTA​TTT​AC-3′ 
RasYQ68L ΔCAAX pGBKT7 F: 5′-CGG​AAT​TCA​TGA​CAA​CAA​ATA​AAA​GTA​ATG​G-3′ 
R: 5′-CGG​GAT​CCT​TAA​ATT​GAT​ATT​TTG​GCA​CCA​TTT​ACT​TTT​TG-3′ 
RasZQ67L ΔCAAX pGBKT7 F: 5′-CGG​AAT​TCA​TGG​CAT​CAT​ATA​AAA​ATA​ATA​AT-3′ 
R: 5′-CGG​GAT​CCT​TAA​ATT​GAG​GTT​TTA​GTA​TTA​TTT​AC-3′ 
RanAQ65L pGBKT7 F: 5′-GGC​ATA​TGG​CAG​AAA​AAG​AAC​AAA​TTA​AAT​TAG-3′ 
R: 5′-GCG​GAT​CCT​TAC​AAG​TCA​TCA​TTG​TCT​TCT​GG-3′ 
RapAQ65E ΔCAAX pGBKT7 F: 5′-GGC​ATA​TGC​CTC​TTA​GAG​AAT​TCA​AAA​TCG-3′ 
R: 5′-CGG​GAT​CCT​TAT​TTT​GAT​TTA​GCT​TTG​CTT​GG-3′ 
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