Table 1.

Cloning Oligonucleotides and Plasmids Used to Produce S5H Proteins

Cloning oligonucleotides	Sequence (restriction sites bolded and underlined)
7nt-NdeI-nagAa-FOR	AGAGTCACATATGATGGAACTGGTAGTAGAACCCCTCAATTTGC
7nt-nagAa-SpeI-REV	AGAGTCAACTAGTTCAGACGCCGCTGGGATAGAACG
7nt-NdeI-nagAb-FOR	AGAGTCACATATGATGACTCAGAACTGGATTGATGCAGC
7nt-nagAb-SpeI-REV	AGAGTCAACTAGTTTACTCCATTTTGAGCATCACGCG
NdeI nagG for2	TGCATATGATGAGTGAACCCCAACGATTAAAACCCGTGTTTCCCC
HindIII nagH rev2	TCAAGCTTTCAGATTGGATAGATCACAGAGTTGGGGATCATTTCGC
Plasmids	Description
pWWF6	pUC18 containing Ralstonia sp. strain U2 nag genes58, 64
pACYC-isc	pACYC-Duet containing part of the E. coli isc operon /P15A origin / chloramphenicol resistant65
p11	pET15b-based plasmid with N-terminal His6-tag and TEV protease site/ ColE1 origin / ampicillin resistanta
pET28b(+)	pET plasmid with N-terminal His6-tag and thrombin cleavage site/ pBR322 origin / kanamycin resistantb
pS5HR	p11 plasmid containing Ralstonia sp. strain U2 nag genes nahAa
pS5HF	p11 plasmid containing Ralstonia sp. strain U2 nag genes nahAb
pS5HH	pET28 plasmid containing Ralstonia sp. strain U2 nag genes nagGH

^aDNASU Plasmid Repository

^bNovagen, Madison, USA