TABLE 1.
Primers used in this study
| Primer | Sequence (5′-3′)a | Restriction enzyme site |
|---|---|---|
| P1 | CCGAATTCGTTAAATCCAAAAGCATACATATATCAATGATGC | EcoRI |
| P2 | CCCGGGGCGGCCCCAAGGTCGAGAACTGAGTTG | SmaI |
| P3 | CCCGGGAGGAGTGACCGACTACTGCGTGAAGAAG | SmaI |
| P4 | GGTCTAGAGTATGATGCAGAAAATATTAAGG | XbaI |
| P5 | CCTCTAGACCAACCATGTTTCCCATGCAATTCACC | XbaI |
| P6 | CCGCGGCCGCATTGAAAATTTTAAAAACCAATAGATGACTCA | NotI |
| 5′RIBO | GGATCCTGGATTTATCCTGATGAGTCCGTGAGGACGAAACTATAGGAAAGGAATTCCTA TAGTCGATAAATCCAAAAGC | BamHI |
| 3′RIBO | GCCGGCGGAAGGGTTAGCTGTGAGATTTTGCATCATTGATATATGTATGCTTTTGGATT TATCGACTATAGGAATTCCTT | NaeI |
| 5′SanDI | CCTCGTCAGCGGGACCCATAATGCC | SanDI |
| 3′ΔXbaIBlpI | CCGCTGAGCGGTTGGTTGAGAGTATGATGC | BlpI |
| 5′GFP | CCAACCGCTGAGCATGGTGAGCAAGGGCGAGG | BlpI |
| 3′GFP | GTGGCTAACGGCAGGTGATTCACGCTTAAGCTCGAGATCTGAGTCCG | |
| 5′nsp4 | GCGTGAATCACCTGCCGTTAGCCACAATGGCGATGGCCACGCTCG | |
| 3′Jun | CCATGCTGAGCGGTTGGTTGAGAGTATGATGC | BlpI |
| nsP4-F | GGCGGCTTCCTGTTACTCGACACGG | |
| GFP-R | TTAAGCTCGAGATCTGAGTCCGG | |
| 5′ProGFP | ATCGATGAACGATATCGGCCGCCGCTACACGCTATGGCG | EcoV |
| 3′ProGFP | CCGGAATGCTAGCTTAAGCTCGAGATCTGAGTCCG | NheI |
| 3′UTR | CGAGCTTAAGCTAGCATTCCGGTATACAAATCGC | NheI |
| T7t | GGCTAGGTCGGCGGCCGCAAAAAACCCCTCAAGACCCG | NotI |
| GSP1 | CCGCCGAGTCGCTCCAGTTGGCG | |
| GSP2 | CGGGTTCTCCAGGACGTCCTTCAAG | |
| 5′RACEseq | GGCGGCGGCATGGTCGTTGGACGACCGG | |
| Cap-R | CCTTCAGCATAGTCATGGCCTTCTTTGG |
a
Restriction enzyme sites are underlined. The italic sequence is part of the nsP4 sequence, and the bold sequence is part of the GFP sequence.