TABLE 2.
Primers and amplicons
| Primer or amplicon | Strand | Positiona | Sequence or modificationb |
|---|---|---|---|
| Primers | |||
| A | + | 400-418 | 5′-GCCGATGTGCCGCCTGTGG-3′ |
| B | − | 1743-1760 | 5′-CCGGGATTACTTGGCAGC-3′ |
| C | + | 2279-2291 | 5′-CATAACCATAGTTGATGCGAG-3′ |
| D | − | 2840-2857 | 5′-CATAAATTCAGGCATAAGG-3′ |
| E | + | 3013-3034 | 5′-GGTCAGATTAATCGTCGTGGC-3′ |
| F | − | 3630-3648 | 5′-GGTGGTGAAACAGGACTGC-3′ |
| G | − | 1069-1089 | 5′-AAAAAAGAATTCTAACACAGCCGAGATAAAAAAG-3′b |
| H | + | 1092-1110 | 5′-TTTTTTGAATTCTAACCCATTTGCTGTCCACCAGTCATGCTAGCCATTTTACAAGTGAAATTCTCG-3′b |
| Amplicons | |||
| AB | 400-1760 | ||
| CD | 2279-2857 | ||
| ABCD | 400-2857 | 518 bp deleted between positions 1760 and 2279 (residues 956 to 1475 in hcaG) | |
| EF | 3013-3630 | ||
| CDEF | 2279-3630 | 115 bp deleted between positions 2857 and 3013 (residues 230 to 386 in hcaF) | |
| AG | 400-1089 | Contains poly(A) leader and EcoRI site | |
| HB | 1092-2297 | Contains poly(T) leader and EcoRI site | |
| AGHB | 400-2297 | Encodes T7 · TAG fused to C terminus of hcaG |
a
Position relative to XhoI site in pobA.
b
The chromosomal sequence is indicated by boldface type, the EcoRI sequence is italicized, and the T7·TAG sequence is underlined.