Primer

Sequence

Digestion site

P1(otrA-F) P2(otrA-R)

5'  CGCCATATGATGAACAAGCTGAATCTGGG  3' 5'  GGAAGCTTTCTAGATCACACGCGCTTGAGC  3'

NdeI XbaI

P3(otrB-F)

5'  CGCCATATGGTGTCATCCGCAAATCCG  3'

NdeI

P4(otrB-R)

5'  CCAAGCTTGCTCTAGATCAGGCGTCCGACGC  3'

XbaI

P5(attB-F)

5’  GTTCACCAACAGCTGGAGGC  3’

P6(attB-R)

5’  CGTCATGCCCGCAGTGACC  3’