Table 1.

Primers, vectors, and kanamycin resistance cartridges used to inactivate genes in Synechocystis PCC 6803

ORF	Primer	Enzyme	Vector	KmR cassette	Insertion site
slr0114	5′; 5′-CTAAGCGGTACCGAATGTCC-3′	KpnI	pBlueIIKS	KIND	NheI
	3′; 5′-GCTAACATGAGCTCCACGTC-3′	SacI
slr1983	5′; 5′-CATCTAGCCCGGGCATATCA-3′	XmaI	pBlueIIKS	KAPA	EcoRI
	3′; 5′-CCAAGAAAATGGTACCAC-3′	KpnI
sll0602	5′; 5′-GGCCATGGTACCGCTTTTATCG-3′	KpnI	pBlueIIKS	KAPA	ApaI
	3′; 3′-GCTAGCGGAGCTCTCATCTAAAC-3′	SacI
sll1033	5′; 5′-GGGGATGGTGTTCTAGAATTAGCG-3′	XbaI	pUC19	KISS	KpnI
	3′; 5′-CCTACCGAGCTCGTGGAAACAG-3′	SacI
sll1365	5′; 5′-GCTGGTACCATTGATTTTGCCC-3′	KpnI	pBlueIIKS	KIXX	SmaI
	3′; 5′-CCTAGAGCTCTACGGCAGTG-3′	SacI
sll1771	5′; 5′-CGATCAAGCTTTTCCGTTTCC-3′	HindIII	pUC19	KISS	KpnI
	3′; 5′-TTACCACTGGCGGAGCTCTCAT-3′	SacI
sll1387	5′; 5′-GCCTACTGGCACCACCAAAA-3′	HincII	pBlueIIKS	KIXX	BamHI
	3′; 5′-GGTCGAAACTTCCGCAAGCT-3′	HindIII
sll1329	5′; 5′-GCCATTCCCAAGGGAGAAAAAG-3′	XmaI	pUC19	KIXX	(SfiI) blunt/SmaI
	3′; 5′-GCACGTAGAGCAAACTGAATTC-3′	SalI
sll1770	5′; 5′-CCGGTACCGTGTTCACAGCGGGCATTATG-3′	KpnI	pBlueIIKS	KIXX	SmaI
	3′; 5′-GCTCTAGACGTCCCTCTGGATCAAGGTA-3′	XbaI

The primers used to PCR amplify the corresponding open reading frames contained unique restriction sites, indicated in bold-face letters, for cloning the fragments into pBluescript II KS (Stratagene) or pUC 19 (Amersham Pharmacia) as indicated. The open reading frames of the resulting constructs were disrupted by inserting into unique restriction sites a kanamycine resistance (Km^R) cartridge derived from either of the pUC4-KIND, pUC4-KAPA, pUC4-KISS or pUC4-KIXX plasmids (Amersham Pharmacia) as indicated.