Table 1.

Primers for site-directed mutagenesis of the wild-type expression plasmid (pcDNA5/FRT-WT-F13A)

MutationSequence 5′→ 3′
Arg100Leu Forward primer: 5′- AGA​AGG​GAT​CTC​TTCCTGGTG​GAA​TAC​GTC​AT −3′ 
Reverse primer: 5′- ATG​ACG​TAT​TCC​ACCAGGAA​GAG​ATC​CCT​TCT −3′ 
Arg100Gln Forward primer: 5′- AGA​AGG​GAT​CTC​TTCCAGGTG​GAA​TAC​GTC​AT −3′ 
Reverse primer: 5′- ATG​ACG​TAT​TCC​ACCTGGAA​GAG​ATC​CCT​TCT −3′ 
Glu111Leu Forward primer: 5′-GGT​CGC​TAC​CCA​CAGCTGAAC​AAG​GGA​ACC​TA-3′ 
Reverse primer: 5′- TAG​GTT​CCC​TTG​TTCAGCTG​TGG​GTA​GCG​ACC-3′ 
Asn112Val Forward primer: 5′- CGC​TAC​CCA​CAG​GAGGTCAAG​GGA​ACC​TAC​AT −3′ 
Reverse primer: 5′- ATG​TAG​GTT​CCC​TTGACCTC​CTG​TGG​GTA​GCG −3′ 
Lys113Leu Forward primer: 5′- TAC​CCA​CAG​GAG​AACCTGGGA​ACC​TAC​ATC​CC −3′ 
Reverse primer: 5′- GGG​ATG​TAG​GTT​CCCAGGTT​CTC​CTG​TGG​GTA −3′ 
Lys257Leu Forward primer: 5′- AGA​GGG​AAT​CCC​ATCCTAGTC​AGC​CGT​GTG​GG −3′ 
Reverse primer: 5′- CCC​ACA​CGG​CTG​ACTAGGAT​GGG​ATT​CCC​TCT −3′ 
Lys257Glu Forward primer: 5′- AGA​GGG​AAT​CCC​ATCGAAGTC​AGC​CGT​GTG​G −3′ 
Reverse primer: 5′- CCA​CAC​GGC​TGA​CTTCGAT​GGG​ATT​CCC​TCT −3′ 
Asp343Val Forward primer: 5′- ATT​TCT​CTG​CCC​ATGTTAAT​GAT​GCC​AAT​TT −3′ 
Reverse primer: 5′- AAA​TTG​GCA​TCA​TTAACATG​GGC​AGA​GAA​AT −3′ 
Glu401Leu Forward primer: 5′- GAC​AGC​ACC​CCC​CAGCTAAAT​AGC​GAT​GGC​AT −3′ 
Reverse primer: 5′- ATG​CCA​TCG​CTA​TTTAGCTG​GGG​GGT​GCT​GTC −3′ 
Asp404Val Forward primer: 5′- CCC​AGG​AAA​ATA​GCGTTGGC​ATG​TAT​CGG​TG −3′ 
Reverse primer: 5′- CAC​CGA​TAC​ATG​CCAACGCT​ATT​TTC​CTG​GG −3′ 
MutationSequence 5′→ 3′
Arg100Leu Forward primer: 5′- AGA​AGG​GAT​CTC​TTCCTGGTG​GAA​TAC​GTC​AT −3′ 
Reverse primer: 5′- ATG​ACG​TAT​TCC​ACCAGGAA​GAG​ATC​CCT​TCT −3′ 
Arg100Gln Forward primer: 5′- AGA​AGG​GAT​CTC​TTCCAGGTG​GAA​TAC​GTC​AT −3′ 
Reverse primer: 5′- ATG​ACG​TAT​TCC​ACCTGGAA​GAG​ATC​CCT​TCT −3′ 
Glu111Leu Forward primer: 5′-GGT​CGC​TAC​CCA​CAGCTGAAC​AAG​GGA​ACC​TA-3′ 
Reverse primer: 5′- TAG​GTT​CCC​TTG​TTCAGCTG​TGG​GTA​GCG​ACC-3′ 
Asn112Val Forward primer: 5′- CGC​TAC​CCA​CAG​GAGGTCAAG​GGA​ACC​TAC​AT −3′ 
Reverse primer: 5′- ATG​TAG​GTT​CCC​TTGACCTC​CTG​TGG​GTA​GCG −3′ 
Lys113Leu Forward primer: 5′- TAC​CCA​CAG​GAG​AACCTGGGA​ACC​TAC​ATC​CC −3′ 
Reverse primer: 5′- GGG​ATG​TAG​GTT​CCCAGGTT​CTC​CTG​TGG​GTA −3′ 
Lys257Leu Forward primer: 5′- AGA​GGG​AAT​CCC​ATCCTAGTC​AGC​CGT​GTG​GG −3′ 
Reverse primer: 5′- CCC​ACA​CGG​CTG​ACTAGGAT​GGG​ATT​CCC​TCT −3′ 
Lys257Glu Forward primer: 5′- AGA​GGG​AAT​CCC​ATCGAAGTC​AGC​CGT​GTG​G −3′ 
Reverse primer: 5′- CCA​CAC​GGC​TGA​CTTCGAT​GGG​ATT​CCC​TCT −3′ 
Asp343Val Forward primer: 5′- ATT​TCT​CTG​CCC​ATGTTAAT​GAT​GCC​AAT​TT −3′ 
Reverse primer: 5′- AAA​TTG​GCA​TCA​TTAACATG​GGC​AGA​GAA​AT −3′ 
Glu401Leu Forward primer: 5′- GAC​AGC​ACC​CCC​CAGCTAAAT​AGC​GAT​GGC​AT −3′ 
Reverse primer: 5′- ATG​CCA​TCG​CTA​TTTAGCTG​GGG​GGT​GCT​GTC −3′ 
Asp404Val Forward primer: 5′- CCC​AGG​AAA​ATA​GCGTTGGC​ATG​TAT​CGG​TG −3′ 
Reverse primer: 5′- CAC​CGA​TAC​ATG​CCAACGCT​ATT​TTC​CTG​GG −3′ 

The mutated codons are underlined.

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