. 2017 Nov 1;2(6):e00306-17. doi: 10.1128/mSphere.00306-17

TABLE 5 .

Primers used in the study

Primer purpose	Gene (GenBank accession no.)	Primer name	Sequence (5′ to 3′)
Detection of kanamycin resistance genes	aphA1 (AF093572)	aphA1-F	CGTCAGCCAGTTTAGTCTG
Detection of kanamycin resistance genes		aphA1-R	CAATCAGGTGCGACAATC
	aphA2 (U66885)	aphA2-F	GGCTATTCGGCTATGACTG
		aphA2-R	GGACAGGTCGGTCTTGAC
	aadB (JN119852)	aadB-F	TACTATGCCGATGAAGTACC
		aadB-R	CAAGACCTCAACCTTTTCC
Amplification of zeo in pCR-Blunt	zeo	oBBI 465	CGATAGCTGAATGAGTGACGTGCTGAAGTTCCTATACTTTCTAGAGAATAGGAACTTCGCTTACAATTTCCTGATGC^a
Amplification of zeo in pCR-Blunt		oBBI 466	GCATAGAGCAGTGACGTAGTCGCGAAGTTCCTATTCTCTAGAAAGTATAGGAACTTCACGTGTCAGTCCTGCTCC^a
Synthesis of an aphA1 knockout fragment by amplification of oBBI 465/466-zeo	aphA1	oBBI 467	CAAGATAAAAATATATCATCATGAACAATAAAACTGTCTGCTTACGATAGCTGAATGAGTGACGTGC^b
		oBBI 468	GGATACTGCCAGATATCGGCGTCCAGGGAGATGTCCTGAAGCATAGAGCAGTGACGTAGTCGC^b
Synthesis of a hilA knockout fragment by amplification of oBBI 92/93-neo	hilA	oBBI 490	GAATACACTATTATCATGCCACATTTTAATCCTGTTCCTGTATCGATAGCTGAATGAGTGACGTGC^c
		oBBI 491	CAAATATTGTCTTCGTTTTTAAATTTATTCCACATTTTCTCGGCATAGAGCAGTGACGTAGTCGC^c

^{^a}

The 5′ end contains a universal binding region and FRT sites. The underlined sequences bind pCR-Blunt to amplify zeo.

^{^b}

Boldface indicates homology to aphA1 for gene knockout. Underlined sequences bind oBBI 465/466-zeo.

^{^c}

Boldface indicates homology to hilA for gene knockout. Underlined sequences bind oBBI 92/93-neo.