TABLE 2.
Primers used for cloning of expression plasmids and mutagenesis of the 3′UTR plasmids
Primer | Sequence (5′–3′)a |
---|---|
ORF31-33-Mutant1-F | CTCGAATTCAGAGACGTCAACCCTTTTGTTTGGCTC |
ORF31-33-Mutant1-R | GAGCCAAACAAAAGGGTTGACGTCTCTGAATTCGAG |
ORF31-33-Mutant2-F | CCACGCACGTCGAAGACGTCCTAACAGGAGTGCTC |
ORF31-33-Mutant2-R | GAGCACTCCTGTTAGGACGTCTTCGACGTGCGTGG |
ORF71-73-Mutant1-F | TATCTGATTTAATAAACTGATACAAGTTTTGTAAGAA |
ORF71-73-Mutant1-R | TTCTTACAAAACTTGTATCAGTTTATTAAATCAGATA |
ORF71-73-Mutant2-F | CCATCACACTCCCAACACTATCGCCATACACCATAGA |
ORF71-73-Mutant2-R | TCTATGGTGTATGGCGATAGTGTTGGGAGTGTGATGG |
ORF71-73-Mutant3-F | GCTGGGGGGCTCCCAAGTGGTGGACTTTTGGCACCAC |
ORF71-73-Mutant3-R | GTGGTGCCAAAAGTCCACCACTTGGGAGCCCCCCAGC |
ORF71-73-Mutant4-F | TGGCACCACGAGGTCAAGTGGCTGATTACAAAAGCC |
ORF71-73-Mutant4-R | GGCTTTTGTAATCAGCCACTTGACCTCGTGGTGCCA |
3Flag-ORF30-F | AAAGGGCCCTGGGTGAGCCAGTGGATCCTGGACA |
3Flag-ORF31-F | AAAGGGCCCTGTCACAAAACAGAAAGACTCTGCCT |
3Flag-ORF32-F | AATGGGCCCTGGATGCGCATGCTATCAACGAAAGA |
3Flag-ORF33-F | ATAGGGCCCTGGCTAGCCGGAGGCGCAAACTTC |
3Flag-ORF30-33-Universal-R | GCCAAGCTTTTAAGCAGTACTTCGGTTTATTTGA |
3Flag-ORF73a-F | TATGCTAGCGGCGCCCCCGGGAATGCGCCTGAGGT |
3Flag-ORF73a-R | AGTACGCTAGCCGTTGTCCGTGTGTTCA |
3Flag-ORF73b-F | TGAACACACGGACAACGGCTAGCGTACT |
3Flag-ORF73b-F | CCCCTCGAGTTCTTACAAAACTTGTTAGTGTTTATTAAATCAGA |
3Flag-Luc-F | GCGGCTAGCGGAAGACGCCAAAAACATAAAGAAAG |
3Flag-Luc-F | GCGGAATTCCGACTCTAGAATTACACGGCGATCT |
ORF30-ApaI-F | AAAGGGCCCTGGGTGAGCCAGTGGATCCTGGACA |
ORF33-HindIII-R | GCCAAGCTTTTAAGCAGTACTTCGGTTTATTTGA |
ORF31-ApaI-F | AAAGGGCCCTGTCACAAAACAGAAAGACTCTGCCT |
ORF32-ApaI-F | AATGGGCCCTGGATGCGCATGCTATCAACGAAAGA |
ORF33-ApaI-F | ATAGGGCCCTGGCTAGCCGGAGGCGCAAACTTC |
Mutated nucleotides and restriction sites are indicated by underlining.