Table 1.
Primer sequences for amn<>cat recombination.
| primers | sequence |
|---|---|
| 5′-amn-cata | 5′-TGTGACATACTATCGGATGTGCGGTAATTGTATGGAACAGGAGACACACTGTGACGGAAGATCACTTCGC |
| 3′-amn-cata | 5′-TACTTACTCCTTCACATCTATTTCTGATAAGAAGGTTCAGAACTTATTAACCAGCAATAGACATAAGCG |
| 5′-catb | 5′-TGTGACGGAAGATCACTTCG |
| 3′-catb | 5′-ACCAGCAATAGACATAAGCG |
| 5′-amnc | 5′-ACAATTCCCTGTTTTGCCTGGC |
| 3′-amnc | 5′-CCCACTGGAATGATCTCCTCG |
a
Retargeting primers contained homologous sequence to E. coli K12 genes upstream and downstream of amn. Bold sequences are homologous to cat.
b
Primers were created within the chloramphenicol gene to verify cat recombination.
c
Verification primers were created upstream and downstream of retargeting primers to ensure proper placement of the cat recombination event within the DY330 genome.