Supplemental Table 1

Primers used for cloning, expression, and gene targeting plasmids

Primer / Sequence / Use
sk gene cloning / Cloning of the full-length sk gene from all GAS strains used
skF / 5'-GGGGATTGAAACTTA-ACTTTTAGG / Forward Primer
skR / 5'-CTGTTGATAGCATGTTG-CTTGC / Reverse primer
sk expression / PCR amplification and cloning of the sk gene into pET42a.
skF1 / 5'-ATTGCTGGGTATGAATGG / Forward primer for SKNS210, SKNS53, SKNZ131, SKAP53, SKNS455, and SKNS88.2
skF2 / 5'-ATTGCTGGACCTGAATGG / Forward primer for SKNS931 and SKNS223
skR1 / 5'-GCGCATGATCGGATCCTT-ATTTGTCTTTAGGGTT / Reverse primer for SKNS210, SKNS53, SKNZ131, SKNS223, and SKNS88.2. The BamH1 site is underlined
skR2 / 5'-GCGCATGATCGGATCCTT-ATTTGTCGTTGGGGTTA / Reverse primer for SKNS931. The BamH1 site is underlined.
skR3 / 5'-GCGCATGATCGGATCCTT-ATTTGTCGTTAGGGTTATC / Reverse primer for SKAP53 and SKNS455. The BamH1 site is underlined
PAMAP53 expression / PCR amplification and cloning of the AP53 pam gene into pET28a
pamF / 5'-GCGCATGATCCCATGGCAAATA-GAGCAGACGACGC / Forward primer. The NcoI site is underlined
pamR / 5’-GCGCATGATCGAATTCTCAG-TGGTGGTGGTGGTGGTGCTGT-CTCTTAGTTTCCTTC / Reverse primer. The EcoR1 site is underlined. The hexahistidine-coding nucleotides are in bold
M1SF370 expression / PCR amplification and cloning of the SF370 emm1 gene into pET28a
emm-1F / 5’-GCGCATGATCCCATGGCAA-ACGGTGATGGTAATCC / Forward primer. The NcoI site is underlined
emm-1R / 5’-GCGCATGATCGAATTCTCA-GTGGTGGTGGTGGTGGTGCT-GTCTCTTAGTTTCCTTC / Reverse primer. The EcoR1 site is underlined. The hexahistidine-coding nucleotides are in bold
AP53/Dpam cells / Inactivation of the pam gene in AP53 cells
pam5F-NotI / 5'-CGCGGCCGCAGTGATCGCA-TCTGCAGGCGTA / Forward primer for the 281 bp upstream sequence of the pam gene containing a Not1 restriction site (underlined)
pam5R / 5'-CCAGTGATTTTTTTCTCCAT-TATTTTATTTGCTCCTTATTTTTTC / Reverse primer for the 281 bp upstream pam-gene. The 5’-ORF cat nucleotides are in bold
pam3F / 5'-AGTGGCAGGGCGGGGCGTA-AGCTATCACTTTGTAATACTG / Forward primer for 282 bp downstream sequence of the pam-gene, 3’-ORF cat nucleotides are in bold
Pam3R-SalI / 5'-CGTCGACAACGGCTACTGAA-GCCGTACCGG-3' / Reverse primer for the 282 bp downstream sequence of the pam gene containing a SalI restriction site (underlined)