TABLE 1.
Design of the MHV-A59 junction sequencesa
| MHV-A59 subclone | Restriction site junction | Location | Junction |
|---|---|---|---|
| A | 5′←TCGTTAA↓CAAATGAGACGNNNN-3′ | 3′-end A fragment, Esp3I, nt 4882 | A/B |
| 3′←AGCAATTGTTT↑ACTCTGCNNNN-5′ | |||
| B | 5′-NNNNCGTCTCA↓CAAATTTGAT→3′ | 5′-end B fragment, Esp3I, nt 4883 | A/B |
| 3′-NNNNGCAGAGTGTTT↑AAACTA→5′ | |||
| 5′←CTTTGCCT↓TAACGGCGAGTTC-3′ | 3′-end B fragment, BglI, nt 9555 | B/C | |
| 3′←GAAACGGAATT↑GCCGCTCAAG-5′ | |||
| C | 5′-CTTTGCCT↓TAACGGCGAGTTC→3′ | 5′-end C fragment, BglI, nt 9556 | B/C |
| 3′-GAAACGGAATT↑GCCGCTCAAG→5′ | |||
| 5′←TAGCATA↓AACCAGAGACGNNNN-3′ | 3′-end C fragment, Esp3I, nt 11510 | C/D | |
| 3′←ATCGTATTTGG↑TCTCTGCNNNN-5′ | |||
| D | 5′-NNNNCGTCTCA↓AACCACGACGTC→3′ | 5′-end D fragment, Esp3I, nt 11511 | C/D |
| 3′-NNNNGCAGAGTTTGG↑TGCTGCAG→5′ | |||
| 5′←TATGCTAT↓CCTAAGAGACGNNNN-3′ | 3′-end D fragment, Esp3I, nt 12961 | D/E | |
| 3′←ATACGATAGGAT↑TCTCTGCNNNN-5′ | |||
| E | 5′-NNNNCGTCTCT↓CCTAAGTGACTG→3′ | 5′-end E fragment, Esp3I, nt 12962 | D/E |
| 3′-NNNNGCAGAGAGGAT↑TCACTGAC→5′ | |||
| 5′←ACTCTAAT↓GTCTAGAGACGNNNN-3′ | 3′-end E fragment, Esp3I, nt 15754 | E/F | |
| 3′←TGAGATTACAGA↑TCTCTGCNNNN-5′ | |||
| F | 5′-NNNNCGTCTCT↓GTCTATCGTGCG→3′ | 5′-end F fragment, Esp3I, nt 15755 | E/F |
| 3′-NNNNGCAGAGACAGA↑TAGCACGC→5′ | |||
| 5′←ACAGTCGG↓TCCGAGAGACGNNNN-3′ | 3′-end F fragment, Esp3I, nt 22739 | F/G | |
| 3′←GTCAGCCAGGC↑TCTCTGCNNNN-5′ | |||
| G | 5′-NNNNCGTCTCG↓TCCGACTGTACC→3′ | 5′-end G fragment, Esp3I, nt 22740 | F/G |
| 3′-NNNNGCAGAGCAGGC↑TGACATGG→5′ |
a
Horizontal arrows indicate the direction of the MHV coding sequences in an individual clone, vertical arrows indicate the site of restriction, and an underline represents the resulting 3- or 4-nucleotide overhang generated at each junction. Various restriction sites are shown in boldface. N, any nucleotide can be inserted at this position. The 3′ end of MHV G is as described for pMH54 (27). nt, nucleotide.