TABLE 2.

Primers for BAC mutagenesis

BAC	Primers
UL31(dN)	5′-ATC TCG CTC CTG TCC CTG GAG CAC ACC CTG TGT ACC TAT GCC GCC TCA CGC CCG CAA ACA TAG GGA TAA CAG GGT-3′, 5′-GCT CGT GTA AAC ACA GCT CCT GTT TGC GGG CGT GAG GCG GCA TAG GTA CAC AGG GTG TGC GCC AGT GTT ACA ACC-3′
UL31(SA3)	5′-TCG ATC TCG CTC CTG TCC CTG GAG CAC ACC CTG TGT ACC TAT GTA TGA CAC CGA CCC CAT CGC CGC GGC GCC CGG CCC GGG CCC TAT CAC GGC AAG GAG CGC CGG CGG GCG CGC TCC TCT GCG GCC GGC GTA GGG ATA ACA GGG T-3′, 5′-GGC CCC GAT AGC GCT GGC GCT CGT GTA AAC ACA GCT CCT GTT TGC GGG CGT GAG GCG GCA GGC TCT TCC GGG CGG CCC GAC GCA CCA CGC CCA GAG TCC CGC CGG CCG CAG AGG AGC GCG CCC GCC GGC GCT CCT TGC CAG TGT TAC AAC C-3′
UL31(SA6)	5′-GCC CGG GCC CTA TCA CGG CAA GGA GCG CCG GCG G GCG CGC GCT GCT GCG GCC GGC GGG ACT CTG GGC GTG GTG CGT CGG GCC GCC CGG AAG TAG GGA TAA CAG GGT-3′, 5′-CGT GTA AAC ACA GCT CCT GTT TGC GGG CGT GAG GCG GCA GTG CCT TCC GGG CGG CCC GAC GCA CCA CGC CCA GAG TCC CGC CGC CAG TGT TAC AAC C-3′
UL31(SAR)	5′-GGG CCT CCC GGA AGA GCC TGC CGC CTC ACG CCC GCA AAC AGG AGC TGT GTT TAC ACG AGC TAG GGA TAA CAG GGT-3′, 5′-GCT CGT GTA AAC ACA GCT CCT GTT TGC GGG CGT GAG GCG GCA TAG GTA CAC AGG GTG TGC GCC AGT GTT ACA ACC-3′b
UL31(SAE)	5′-CCC GGG CCC TAT CAC GGC AAG GAG CGC CGG CGG GCG CGC GAA GAA GCG GCC GGC GGG ACT CTG GGC GTG GTG CGT CGG GCC GCC CTA GGG ATA ACA GGG T-3′, 5′-GTG TAA ACA CAG CTC CTG TTT GCG GGC GTG AGG CGG CAG TTC CTT CCG GGC GGC CCG ACG CAC CAC GCC CAG AGT CCC GCC GGC CGC CAG TGT TAC AAC C-3′
UL31(SE6)	5′-TCG ATC TCG CTC CTG TCC CTG GAG CAC ACC CTG TGT ACC TAT GTA TGA CAC CGA CCC CAT CGC CGC GGC GAA CGG CCC GGG CCC TAT CAC GGC AAG GAG CGC CGG CGG GAA CGC GAAGAA GCG GCC GGC GTA GGG ATA ACA GGG T-3′, 5′-GGC CCC GAT AGC GCT GGC GCT CGT GTA AAC ACA GCT CCT GTT TGC GGG CGT GAG GCG GCA GTT CCT TCC GTT CGG CCC GAC GCA CCA CGC CCA GAG TCC CGC CGG CCG CTT CTT CGC GTT CCC GCC GGC GCT CCT TGC CAG TGT TAC AAC C-3′
UL31(SE6)/US3(K220A)	5′-TGA CAG CAG CCA TCC AGA TTA CCC CCA ACG GGT AAT CGT GGC GGC GGG GTG GTA CAC GAG CAT AGG GAT AAC AGG GTA ATC GAT TT-3′, 5′-CAG TCG CGC CTC GTG GCT CGT GCT CGT GTA CCA CCC CGC CGC CAC GAT TAC CCG TTG GGG GTG CCA GTG TTA CAA CCA ATT AAC C-3′

aIntroduced mutations are underlined.

^bAmplicons generated using these primers were PCR fused with a second amplicon generated from wild-type HSV-1(F) genomic DNA with primer pair 5′-TGCATAGCGTTGCGTGCC-3′ and 5′-TTTGCGGGCGTGAGGC-3′ to obtain the 5′ extension required for homologous recombination.