Table 2.

Primers used for cloning by Gibson assembly

ConstructInsert forward primer (5′ → 3′)Insert reverse primer (5′ → 3′)Vector forward primer (5′ → 3′)Vector reverse primer (5′ → 3′)
pCMV3_FLAG-IMPDH1(546)-WT GAC​TAC​AAA​GAC​GAT​GAC​GAC​AAG​AGC​ATG​GCG​GAC​TAC​CTG​ATC GAG​GCT​GAT​CAG​CGG​GTT​TAA​TTA​TGC​AGC​GAC​GCA​GTC GAC​TGC​GTC​GCT​GCA​TAA​TTA​AAC​CCG​CTG​ATC​AGC​C CTG​ATC​AGG​TAG​TCC​GCC​ATG​CTC​TTG​TCG​TCA​TCG 
pCMV3_FLAG-IMPDH1(546)-S477D GAC​TAC​AAA​GAC​GAT​GAC​GAC​AAG​AGC​ATG​GCG​GAC​TAC​CTG​ATC GAG​GCT​GAT​CAG​CGG​GTT​TAA​TTA​TGC​AGC​GAC​GCA​GTC GAC​TGC​GTC​GCT​GCA​TAA​TTA​AAC​CCG​CTG​ATC​AGC​C CTG​ATC​AGG​TAG​TCC​GCC​ATG​CTC​TTG​TCG​TCA​TCG 
pCMV3_FLAG-IMPDH1(595)-WT GAT​GAC​GAC​AAG​AGC​ATG​GAG​GGG​CCA​CTC​ACT​C GAG​GCT​GAT​CAG​CGG​GTT​TAA​TTA​TGC​AGC​GAC​GCA​GTC GAC​TGC​GTC​GCT​GCA​TAA​TTA​AAC​CCG​CTG​ATC​AGC​C GAG​TGA​GTG​GCC​CCT​CCA​TGC​TCT​TGT​CGT​CAT​C 
pCMV3_FLAG-IMPDH1(595)-S477D GAT​GAC​GAC​AAG​AGC​ATG​GAG​GGG​CCA​CTC​ACT​C GAG​GCT​GAT​CAG​CGG​GTT​TAA​TTA​TGC​AGC​GAC​GCA​GTC GAC​TGC​GTC​GCT​GCA​TAA​TTA​AAC​CCG​CTG​ATC​AGC​C GAG​TGA​GTG​GCC​CCT​CCA​TGC​TCT​TGT​CGT​CAT​C 
pCMV3_FLAG-IMPDH1(546)-S477A CGC​AGC​CTG​GCT​GTC​CTT​CGG​TCC​ATG​ATG TGG​ACC​GAA​GGA​CAG​CCA​GGC​TGC​G   
pCMV3_FLAG-IMPDH1(595)-S477A CGC​AGC​CTG​GCT​GTC​CTT​CGG​TCC​ATG​ATG TGG​ACC​GAA​GGA​CAG​CCA​GGC​TGC​G   
ConstructInsert forward primer (5′ → 3′)Insert reverse primer (5′ → 3′)Vector forward primer (5′ → 3′)Vector reverse primer (5′ → 3′)
pCMV3_FLAG-IMPDH1(546)-WT GAC​TAC​AAA​GAC​GAT​GAC​GAC​AAG​AGC​ATG​GCG​GAC​TAC​CTG​ATC GAG​GCT​GAT​CAG​CGG​GTT​TAA​TTA​TGC​AGC​GAC​GCA​GTC GAC​TGC​GTC​GCT​GCA​TAA​TTA​AAC​CCG​CTG​ATC​AGC​C CTG​ATC​AGG​TAG​TCC​GCC​ATG​CTC​TTG​TCG​TCA​TCG 
pCMV3_FLAG-IMPDH1(546)-S477D GAC​TAC​AAA​GAC​GAT​GAC​GAC​AAG​AGC​ATG​GCG​GAC​TAC​CTG​ATC GAG​GCT​GAT​CAG​CGG​GTT​TAA​TTA​TGC​AGC​GAC​GCA​GTC GAC​TGC​GTC​GCT​GCA​TAA​TTA​AAC​CCG​CTG​ATC​AGC​C CTG​ATC​AGG​TAG​TCC​GCC​ATG​CTC​TTG​TCG​TCA​TCG 
pCMV3_FLAG-IMPDH1(595)-WT GAT​GAC​GAC​AAG​AGC​ATG​GAG​GGG​CCA​CTC​ACT​C GAG​GCT​GAT​CAG​CGG​GTT​TAA​TTA​TGC​AGC​GAC​GCA​GTC GAC​TGC​GTC​GCT​GCA​TAA​TTA​AAC​CCG​CTG​ATC​AGC​C GAG​TGA​GTG​GCC​CCT​CCA​TGC​TCT​TGT​CGT​CAT​C 
pCMV3_FLAG-IMPDH1(595)-S477D GAT​GAC​GAC​AAG​AGC​ATG​GAG​GGG​CCA​CTC​ACT​C GAG​GCT​GAT​CAG​CGG​GTT​TAA​TTA​TGC​AGC​GAC​GCA​GTC GAC​TGC​GTC​GCT​GCA​TAA​TTA​AAC​CCG​CTG​ATC​AGC​C GAG​TGA​GTG​GCC​CCT​CCA​TGC​TCT​TGT​CGT​CAT​C 
pCMV3_FLAG-IMPDH1(546)-S477A CGC​AGC​CTG​GCT​GTC​CTT​CGG​TCC​ATG​ATG TGG​ACC​GAA​GGA​CAG​CCA​GGC​TGC​G   
pCMV3_FLAG-IMPDH1(595)-S477A CGC​AGC​CTG​GCT​GTC​CTT​CGG​TCC​ATG​ATG TGG​ACC​GAA​GGA​CAG​CCA​GGC​TGC​G   
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