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Table 3 PCR primers used to create T4ΩRD mutants and the corresponding genes deleted

From: Regions of Diversity 8, 9 and 13 contribute to Streptococcus pneumoniae virulence

  Genes deleted: TIGR annotation Primers used to amplify flanking DNA fragments for PCR construct deletion mutagenesis†
Strain   Upstream fragment Downstream fragment
T4 ΩRD2 SP0163-0168 SP0162 F: NNNNNaagcttGGGATTGGGCTCCTATG SP0168 F: NNNNNgcggccgcGTACCAACAAATACCTC
   SP0163 R: NNNNNactagtCTGCTAAAGATAGG SP0170 R: NNNNNtctagaTCTAACTCTTCATAG
T4 ΩRD5 SP0694-0700 SP0694 F: NNNNNaagcttCCTATCTTGTTGTCATTA SP0698 F: NNNNNgcggccgcATGGGAACTACTCC
   SP0694 R: NNNNNactagtCGTAGAACATATTTCGTCC SP0800 R:NNNNNtctagaACCCAAAACTTCTGC
T4 ΩRD7 SP1130-1145 SP1130 F: NNNNNaagcttCAATTTCTGAAGGTCCG SP1145 F: NNNNNgcggccgcCATACTGATGATGAGG
   SP1130 R: NNNNNactagtGTTTAAGCCATGGCG SP1146 R: NNNNNtctagaCCATAACTAACTCTCC
T4 ΩRD8 SP1315-1352 SP1315 F: NNNNNgcggccgcACTTCGGAAAGAAGTGG SP1352 F: NNNNNaagcttGTTTCAATGCCCAGCTTCGTCC
   SP1315 R: NNNNNtctagaCTTCACTTTCATAATACG SP1352 R: NNNNNactagtCTTTAATGCATCATTAACGACGC
T4 ΩRD8a SP1315-1331 SP1315 F SP1331 F: NNNNNaagcttTGTTACTGCAAAAAGAAC
   SP1315 R SP1331 R: NNNNNactagtGCGCCTTTATCTGCAGC
T4 ΩRD8b SP1332-1352 SP1332 F: NNNNNgcggccgcGAATTTCTTTGTATCGG SP1352 F
   SP1332 R: NNNNNtctagaGCAGCAAACTATAAAC SP1352 R
T4 ΩRD8b1 SP1332-1337 SP1332 F SP1337 F: NNNNNaagcttCAGTTTACCAAATCATC
   SP1332 R SP1337 R: NNNNNactagtCTGCCTCTTCAGAACAATAACG
T4 ΩRD8b2 SP1338-1344 SP1339 F: NNNNNgcggccgcTGCGGTAACTGGAGTG SP1344 F: NNNNNaagcttTAAAATATACTGGCACGG
   SP1338 R: NNNNNtctagaTTCTTCTCTACAAGCTC SP1344 R: CGATGTAGTTactagtGCC
T4 ΩRD8b3 SP146-1352 SP1346 F: NNNNNgcggccgcCCAACAAGTAAATGTCC SP1352 F
   SP1346 R:NNNNNtctagaTATAGCGCACCATACC SP1352 R
T4 ΩRD9 SP1615-1621 SP1615 F: NNNNNaagcttGGGACTTAACTTGAGC SP1621 F: NNNNNgcggccgcATAGGGAATGTTTACCC
   SP1615 R: NNNNNactagtCCATTCAGGAGTTGCC SP1621 R: NNNNNtctagaAAACAAGAAC
T4 ΩRD12 SP1950-1957 SP1950 F: NNNNNaagcttACGAGTGGATTGAC SP1957 F: NNNNNgcggccgcGATTGAACTTAAACAGG
   SP1950 R: NNNNNactagtCTTCATAGCTGTGATCCG SP1957 R: NNNNNtctagaGCCATCTCCCAAATTGCC
T4 ΩRD13 SP2158-2166 SP2158 F: NNNNNaagcttGGAATTCTCAACATAG SP2166 F: NNNNNgcggccgcCATCTCCTCATCAGG
   SP2158 R: NNNNNactagtCCACAAATCTTTGC SP2166 R: NNNNNtctagaCAGATTTGACGAAAGG