TABLE 2.
Sequences of the primers used in this study
| Primer | Nucleotide sequencea |
|---|---|
| Cloning of tyrA | |
| tyrA3 | 5′ GGCTTAAGAGGTTTACCATGGTTGCTGAATTG 3′ |
| tyrA5 | 5′ CCCCAAGCTTGATGAAAAGGTGCCGGATGATGTG 3′ |
| Cloning of tyrC | |
| tyrC5b | 5′ GCAGGCGCTCTCCATGGCCGTCTTTAAG 3′ |
| tyrC3 | 5′ CCCCAAGCTTGTTCATGCTGCGATCAATCG 3′ |
| Cloning of pheACM | |
| CM5 | 5′ CCCCAAGCTTCACACAGGAAACAGACCATGG 3′ |
| CM3c | 5′ CCCCAAGCTTTTATCAGAGAAAAGCGATGCGTGC 3′ |
| Sequencing | |
| M13 reversed | 5′ CAGGAAACAGCTATGAC 3′ |
| M13 forwardd | 5′ GTAAAACGACGGCCAG 3′ |
| tyrA1500 | 5′ CTGTTTTATCAGACCGCTTCTGC 3′ |
| SectyrA5 | 5′ GTGTGCCAATCCACGTTACTG 3′ |
| pTrc2 | 5′ CAGCTTATCATCGACTGCAC 3′ |
a
In the nucleotide sequences key restriction enzyme sites are underlined.
b
To clone the tyrC gene in pTrc99A, the GTG start codon in the wild-type gene was changed to ATG in order to create an NcoI site.
c
Stop codons are indicated by bold type.
d
Obtained from Invitrogen (Carlsbad, CA).