TABLE 2.

Oligonucleotides used to modify or delete regions of the ERAV 5′-NTR

Oligonucleotide	Mutation	Sequence (5′→3′)c
TH-GFPR	3′ end of GFP	TTG AGC TCG AGA TCA GAG TCC
TH1-omA1Fa	Alter ATG1 to ATA	TAC TGA CAT AAT GGC GGC G
TH1-omA1Rb	Alter ATG1 to ATA	GAC GCC GCC ATT ATG TCA GTA
TH1-omA2Fa	Alter ATG2 to ATA	GTA CTG ACA TGA TAG CGG CGT CTA AG
TH1-omA2Rb	Alter ATG2 to ATA	GAC GCC GCT ATC ATG TCA GTA
TH1-omA1/2Fa	Alter ATG1 and ATG2 to ATA	GTA CTG ACA TAA TAG CGG CGT C
TH1-omA1/2Rb	Alter ATG1 and ATG2 to ATA	GAC GCC GCT ATT ATG TCA GTA
TH1-omA4Fa	Alter ATG4 to ATA	GGT GCC GTT CGC ATG ATA GAC AAA TTC TTG
TH1-omA4Rb	Alter ATG4 to ATA	TCT ATC ATG CGA ACG GCA CCT GC
TH1-omA3/4Fa	Alter ATG3 and ATG4 to ATA	GGT GCC GTT CGC CAT AAT AGA CAA ATT CTT G
TH1-omA3/4Rb	Alter ATG3 and ATG4 to ATA	TCT ATT ATG CGA ACG GCA CCT GC
TH1-omTAFa	ATG2 Kozak modification	CTG ACT TGA TGA CGG CGT CTA AGG
TH1-omTARb	ATG2 Kozak modification	CGC CGT CAT CAA GTC AGT ACA AAA G
TH1-GFPMFa	5′ end of GFP	ATG GTG AGC AAG GGC GAG GAG CTG TTC
TH1-odA1Rb	Delete sequence in pE1(245–961) between ATG1 and GFP ATG	CTC GCC CTT GCT CAC CAT GTC AGT ACA AAA GGA AAA GAA ACG C
TH1-odA2Rb	Delete sequence in pE1(245–961) between ATG2 and GFP ATG	CGC CCT TGC TCA CCA TCA TGT CAG TAC AAA AGG AAA AGA AAC GC
TH1-odA3Rb	Delete sequence in pE1(245–961) between ATG2 and GFP ATG	CGC CCT TGC TCA CCA TGC GAA CGG CAC CTG CCA GAG TCT GCT C
TH1-odA4Rb	Delete sequence in pE1(245–961) between ATG4 and GFP ATG	CGC CCT TGC TCA CAT CAT GCG AAC GGC ACC TGC CAG AGT CTG CTC

^aOligonucleotides used with TH-GFPR for first-round PCR. 

^bOligonucleotides used with TH1-F245 for first-round PCR. Overlapping first-round PCR products were combined and subjected to second-round PCR with TH1-F245 (Table1) and TH-GFPR. 

^cUnderlined nucleotides, mutated bases; boldfaced nucleotides, overlaps with TH1-GFPMF.