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 |
a
Mutated nucleotides and restriction sites are indicated by underlining.