Table 2.

Primer sequences

Primer Sequence (5′ to 3′) 
pLVX-Rap2a forward (used to make GFP WT, G12V, S17N, Eos WT) AAG​GCG​AAT​TCC​GCG​AGT​ACA​AAG​TGG​TGG​TGC​TG 
pLVX-Rap2a reverse (used to make GFP WT, G12V, S17N, Eos WT) AAG​GCT​CTA​GAC​TAT​TGT​ATG​TTA​CAT​GCA​GAA​CA 
pLVX-mEos3.2 forward AAG​GCC​TCG​AGA​TGA​GTG​CGA​TTA​AGC​CAG​AC 
pLVX-mEos3.2 reverse AAG​GCG​AAT​TCT​CGT​CTG​GCA​TTG​TCA​GGC​AA 
Rap2a-S17N forward (mutagenesis) GGT​AGG​CAA​AAA​CGC​CCT​GAC​CGT​GCA​GTT​C 
Rap2a-S17N reverse (mutagenesis) GAA​CTG​CAC​GGT​CAG​GGC​GTT​TTT​GCC​TAC​C 
pLVX-mCherry-Rap2a forward GGA​TCC​CGC​GAG​TAC​AAA​GTG​GTG​GTG​CTG 
pLVX-mCherry-Rap2a reverse TCT​AGA​CTA​TTG​TAT​GTT​ACA​TGC​AGA​ACA 
pLVX-Eos-Rap2b forward AAG​GCG​AAT​TCA​GAG​AGT​ACA​AAG​TGG​TGG​TAC​TG 
pLVX-Eos-Rap2b reverse TCT​AGA​TCA​CAG​GAT​CAC​GCA​GGC​CGA 
pLVX-Eos-Rap2c forward AAG​GCG​AAT​TCA​GGG​AAT​ACA​AGG​TAG​TGG​TGT​TA 
pLVX-Eos-Rap2c reverse TCT​AGA​TTA​CTG​GAC​AAC​ACA​AGT​TGT​ACA 
pGEX-KG-Rap2a forward CCG​GAA​TTC​TAC​GCG​AGT​ACA​AAG​TGG​TGG​TGC​TGG​GC 
pGEX-KG-Rap2a reverse CCG​CTC​GAG​CTA​TTG​TAT​GTT​ACA​TGC​AGA​ACA​GCA​TGG​GTC 
pLVX-GFP-Rab40b forward AAG​GCG​AAT​TCA​TGG​TGA​GCA​AGG​GCG​AGG​AGC 
pLVX-GFP-Rab40b reverse AAG​GCG​GAT​CCT​TAA​GAA​ATT​TTG​CAG​CTG​TTT​CTG​GTG​C 
pGPS-FLAG-Rab40b forward (4A and ΔSOCS) GTT​CCA​GAT​TAC​GCG​GTC​GAC​ATG​AGC​GCC​CTG​GGC​AGC 
pGPS-FLAG-Rab40b reverse (4A and ΔSOCS) GAT​GCA​TGC​TCG​AGC​GGC​CGC​TTA​AGA​AAT​TTT​GCA​GCT​G 
pLVX-Eos-Rap2aK5R forward AAG​GCG​AAT​TCC​GCG​AGT​ACC​GCG​TGG​TGG​TGC​TG 
pLVX-Eos-Rap2aK5R reverse AAG​GCT​CTA​GAC​TAT​TGT​ATG​TTA​CAT​GCA​GAA​CA 
pGEX-KG-Rap2aK5R forward AAG​GCG​AAT​TCT​ACG​CGA​GTA​CCG​CGT​GGT​GGT​GCT​GGG​C 
pGEX-KG-Rap2aK5R reverse CCG​CTC​GAG​CTA​TTG​TAT​GTT​ACA​TGC​AGA​ACA​GCA​TGG​GTC 
pLVX-Eos-Rap2aK3R forward AAG​GCG​AAT​TCC​GCG​AGT​ACA​AAG​TGG​TGG​TGC​TG 
pLVX-Eos-Rap2aK3R reverse AAG​GCT​CTA​GAC​TAT​TGT​ATG​TTA​CAT​GCA​GAA​CA 
pGEX-KG-Rap2aK3R forward CCG​GAA​TTC​TAC​GCG​AGT​ACA​AAG​TGG​TGG​TGC​TGG​GC 
pGEX-KG-Rap2aK3R reverse CCG​CTC​GAG​CTA​TTG​TAT​GTT​ACA​TGC​AGA​ACA​GCA​TGG​GTC 
pGEX-KG-Rap2aK2R forward AAG​GCG​AAT​TCT​ACG​CGA​GTA​CCG​CGT​GGT​GGT​GCT​GGG​C 
pGEX-KG-Rap2aK2R reverse CCG​CTC​GAG​CTA​TTG​TAT​GTT​ACA​TGC​AGA​ACA​GCA​TGG​GTC 
pGEX-KG-Rap2aK117R,K148R,K150R,K148R/K150R forward (individual mutants) CCG​GAA​TTC​TAC​GCG​AGT​ACA​AAG​TGG​TGG​TGC​TGG​GC 
pGEX-KG-Rap2aK117R,K148R,K150R,K148R/K150R reverse (individual mutants) CCG​CTC​GAG​CTA​TTG​TAT​GTT​ACA​TGC​AGA​ACA​GCA​TGG​GTC 
pGEX-KG-RalGDS-788 forward CCG​GAA​TTC​TAG​CGC​TGC​CGC​TCT​ACA​ACC​AGC​AG 
pGEX-KG-RalGDS-885 reverse CCG​CTC​GAG​CTA​GAA​CGT​CCG​CTT​CTT​GAG​GAC​AAA 
pEGFP-C2-Rap2a forward AAG​GCC​TCG​AGC​CGC​GAG​TAC​AAA​GTG​GTG​GTG​CTG 
pEGFP-C2-Rap2a reverse AAG​GCG​AAT​TCC​TAT​TGT​ATG​TTA​CAT​GCA​GAA​CA 
crRNA-Rab40b Exon3 TCC​AGG​GAT​ACT​TCA​GGC​CA 
crRNA-Rab40b Exon5 TCT​GGC​GGC​CGA​GCA​AGG​GT 
Rab40b-Exon3 genotyping forward CTG​ACT​GGC​GGG​TGT​GTG​GTG​CAC​TTG​CTG 
Rab40b-Exon3 genotyping reverse CCT​CCA​ATC​ATC​CAT​TAA​AGG​ATG​GTA​AGT​TAA​GC 
Rab40b-Exon5 genotyping forward GCT​AGC​ATG​CCC​CCG​GAG​TCC​CCA​AGA​T 
Rab40b-Exon5 genotyping reverse AGG​CAG​ACA​CTT​ATC​CAG​GGG​GCC​GCT​G 
crRNA-Rap2a-Exon1 GAT​GCG​CGA​GTA​CAA​AGT​GG 
crRNA-Rap2b-Exon1 CAT​GAG​AGA​GTA​CAA​AGT​GG 
crRNA-Rap2c-Exon2 GGT​GAA​GGT​GAG​ACT​CAT​GA 
Rap2a-Exon1 genotyping forward GCG​GTG​CTT​GTT​TAT​CTC​CG 
Rap2a-Exon1 genotyping reverse ACG​AGG​CTG​TAG​ACG​AGG​AT 
Rap2b-Exon1 genotyping forward CCT​CCG​TTC​GGT​GGT​TTC​CG 
Rap2b-Exon1 genotyping reverse GAC​GGC​GAC​GAG​TCC​ACC​TC 
Rap2c-Exon2 genotyping forward GTT​GGG​CCC​TCT​GCT​CCT​CT 
Rap2c-Exon2 genotyping reverse GTC​CAG​AAT​TTC​CAG​CAC​GG 
Primer Sequence (5′ to 3′) 
pLVX-Rap2a forward (used to make GFP WT, G12V, S17N, Eos WT) AAG​GCG​AAT​TCC​GCG​AGT​ACA​AAG​TGG​TGG​TGC​TG 
pLVX-Rap2a reverse (used to make GFP WT, G12V, S17N, Eos WT) AAG​GCT​CTA​GAC​TAT​TGT​ATG​TTA​CAT​GCA​GAA​CA 
pLVX-mEos3.2 forward AAG​GCC​TCG​AGA​TGA​GTG​CGA​TTA​AGC​CAG​AC 
pLVX-mEos3.2 reverse AAG​GCG​AAT​TCT​CGT​CTG​GCA​TTG​TCA​GGC​AA 
Rap2a-S17N forward (mutagenesis) GGT​AGG​CAA​AAA​CGC​CCT​GAC​CGT​GCA​GTT​C 
Rap2a-S17N reverse (mutagenesis) GAA​CTG​CAC​GGT​CAG​GGC​GTT​TTT​GCC​TAC​C 
pLVX-mCherry-Rap2a forward GGA​TCC​CGC​GAG​TAC​AAA​GTG​GTG​GTG​CTG 
pLVX-mCherry-Rap2a reverse TCT​AGA​CTA​TTG​TAT​GTT​ACA​TGC​AGA​ACA 
pLVX-Eos-Rap2b forward AAG​GCG​AAT​TCA​GAG​AGT​ACA​AAG​TGG​TGG​TAC​TG 
pLVX-Eos-Rap2b reverse TCT​AGA​TCA​CAG​GAT​CAC​GCA​GGC​CGA 
pLVX-Eos-Rap2c forward AAG​GCG​AAT​TCA​GGG​AAT​ACA​AGG​TAG​TGG​TGT​TA 
pLVX-Eos-Rap2c reverse TCT​AGA​TTA​CTG​GAC​AAC​ACA​AGT​TGT​ACA 
pGEX-KG-Rap2a forward CCG​GAA​TTC​TAC​GCG​AGT​ACA​AAG​TGG​TGG​TGC​TGG​GC 
pGEX-KG-Rap2a reverse CCG​CTC​GAG​CTA​TTG​TAT​GTT​ACA​TGC​AGA​ACA​GCA​TGG​GTC 
pLVX-GFP-Rab40b forward AAG​GCG​AAT​TCA​TGG​TGA​GCA​AGG​GCG​AGG​AGC 
pLVX-GFP-Rab40b reverse AAG​GCG​GAT​CCT​TAA​GAA​ATT​TTG​CAG​CTG​TTT​CTG​GTG​C 
pGPS-FLAG-Rab40b forward (4A and ΔSOCS) GTT​CCA​GAT​TAC​GCG​GTC​GAC​ATG​AGC​GCC​CTG​GGC​AGC 
pGPS-FLAG-Rab40b reverse (4A and ΔSOCS) GAT​GCA​TGC​TCG​AGC​GGC​CGC​TTA​AGA​AAT​TTT​GCA​GCT​G 
pLVX-Eos-Rap2aK5R forward AAG​GCG​AAT​TCC​GCG​AGT​ACC​GCG​TGG​TGG​TGC​TG 
pLVX-Eos-Rap2aK5R reverse AAG​GCT​CTA​GAC​TAT​TGT​ATG​TTA​CAT​GCA​GAA​CA 
pGEX-KG-Rap2aK5R forward AAG​GCG​AAT​TCT​ACG​CGA​GTA​CCG​CGT​GGT​GGT​GCT​GGG​C 
pGEX-KG-Rap2aK5R reverse CCG​CTC​GAG​CTA​TTG​TAT​GTT​ACA​TGC​AGA​ACA​GCA​TGG​GTC 
pLVX-Eos-Rap2aK3R forward AAG​GCG​AAT​TCC​GCG​AGT​ACA​AAG​TGG​TGG​TGC​TG 
pLVX-Eos-Rap2aK3R reverse AAG​GCT​CTA​GAC​TAT​TGT​ATG​TTA​CAT​GCA​GAA​CA 
pGEX-KG-Rap2aK3R forward CCG​GAA​TTC​TAC​GCG​AGT​ACA​AAG​TGG​TGG​TGC​TGG​GC 
pGEX-KG-Rap2aK3R reverse CCG​CTC​GAG​CTA​TTG​TAT​GTT​ACA​TGC​AGA​ACA​GCA​TGG​GTC 
pGEX-KG-Rap2aK2R forward AAG​GCG​AAT​TCT​ACG​CGA​GTA​CCG​CGT​GGT​GGT​GCT​GGG​C 
pGEX-KG-Rap2aK2R reverse CCG​CTC​GAG​CTA​TTG​TAT​GTT​ACA​TGC​AGA​ACA​GCA​TGG​GTC 
pGEX-KG-Rap2aK117R,K148R,K150R,K148R/K150R forward (individual mutants) CCG​GAA​TTC​TAC​GCG​AGT​ACA​AAG​TGG​TGG​TGC​TGG​GC 
pGEX-KG-Rap2aK117R,K148R,K150R,K148R/K150R reverse (individual mutants) CCG​CTC​GAG​CTA​TTG​TAT​GTT​ACA​TGC​AGA​ACA​GCA​TGG​GTC 
pGEX-KG-RalGDS-788 forward CCG​GAA​TTC​TAG​CGC​TGC​CGC​TCT​ACA​ACC​AGC​AG 
pGEX-KG-RalGDS-885 reverse CCG​CTC​GAG​CTA​GAA​CGT​CCG​CTT​CTT​GAG​GAC​AAA 
pEGFP-C2-Rap2a forward AAG​GCC​TCG​AGC​CGC​GAG​TAC​AAA​GTG​GTG​GTG​CTG 
pEGFP-C2-Rap2a reverse AAG​GCG​AAT​TCC​TAT​TGT​ATG​TTA​CAT​GCA​GAA​CA 
crRNA-Rab40b Exon3 TCC​AGG​GAT​ACT​TCA​GGC​CA 
crRNA-Rab40b Exon5 TCT​GGC​GGC​CGA​GCA​AGG​GT 
Rab40b-Exon3 genotyping forward CTG​ACT​GGC​GGG​TGT​GTG​GTG​CAC​TTG​CTG 
Rab40b-Exon3 genotyping reverse CCT​CCA​ATC​ATC​CAT​TAA​AGG​ATG​GTA​AGT​TAA​GC 
Rab40b-Exon5 genotyping forward GCT​AGC​ATG​CCC​CCG​GAG​TCC​CCA​AGA​T 
Rab40b-Exon5 genotyping reverse AGG​CAG​ACA​CTT​ATC​CAG​GGG​GCC​GCT​G 
crRNA-Rap2a-Exon1 GAT​GCG​CGA​GTA​CAA​AGT​GG 
crRNA-Rap2b-Exon1 CAT​GAG​AGA​GTA​CAA​AGT​GG 
crRNA-Rap2c-Exon2 GGT​GAA​GGT​GAG​ACT​CAT​GA 
Rap2a-Exon1 genotyping forward GCG​GTG​CTT​GTT​TAT​CTC​CG 
Rap2a-Exon1 genotyping reverse ACG​AGG​CTG​TAG​ACG​AGG​AT 
Rap2b-Exon1 genotyping forward CCT​CCG​TTC​GGT​GGT​TTC​CG 
Rap2b-Exon1 genotyping reverse GAC​GGC​GAC​GAG​TCC​ACC​TC 
Rap2c-Exon2 genotyping forward GTT​GGG​CCC​TCT​GCT​CCT​CT 
Rap2c-Exon2 genotyping reverse GTC​CAG​AAT​TTC​CAG​CAC​GG 
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