Table 1. Expression plasmid construction.

	EcTet (31–252)	HiMon (33–253)
Source organism	E. coli MG1655	H. influenzae Rd
DNA source	pCB40 plasmid DNA (Paradis-Bleau et al., 2010 ▸)	H. influenzae Rd genomic DNA
Cloning method	Isothermal (NEBuilder HiFi, NEB)	Restriction enzymes
Forward primer	agaacctgtacttccAATCCACTCCCGATCAGTCCACTG†	CATGCCATGGCGAATTTCACGCAAACCTTACAA‡
Reverse primer	cgttatccacttccaatattttaTTTAAACGCTTTTACGTTAACCAAC†	GCCGACGTCGACTTGTTGGAAATTAAGCAATGTAAG§
Forward primer to amplify plasmid	taaAATATTGGAAGTGGATAACGGATC
Reverse primer to amplify plasmid	GGATTGGAAGTACAGGTTCTCG
Cloning vector	pMCSG7 (Stols et al., 2002 ▸)	pETBlue-2 (Novagen)
Expression vector	pMCSG7	pETBlue-2
Expression host	E. coli Origami 2(DE3) (Novagen)	E. coli Tuner(DE3)/pLacI (Novagen)
Complete amino-acid sequence of the construct produced	mhhhhhhssgvdlgtenlyfqSTPDQSTAYMQGTAQADSAFYLQQMQQSSDDTRINWQLLAIRALVKEGKTGQAVELFNQLPQELNDAQRREKTLLAVEIKLAQKDFAGAQNLLAKITPADLEQNQQARYWQAKIDASQGRPSIDLLRALIAQEPLLGAKEKQQNIDATWQALSSMTQEQANTLVINADENILQGWLDLQRVWFDNRNDPDMMKAGIADWQKRYPNNPGAKMLPTQLVNVKAFK¶	MANFTQTLQKDANASSEFYINKLGQTQELEDQQTYKLLAARVLIRENKVEQSAALLRELGELNDAQKLDRALIEARISAAKNANEVAQNQLRALDLNKLSPSQKSRYYETLAIVAENRKDMIEAVKARIEMDKNLTDVQRHQDNIDKTWALLRSANTGVINNASDEGNAALGGWLTLIKAYNDYIRQPVQLSQALQSWKNAYPNHAAATLFPKELLTLLNFQQVEHHHHHH ††

^†The primer 5′ extensions shown in lower case base-pair with sequences on the linearized plasmid for the isothermal cloning procedure.

^‡The NcoI site for cloning is underlined.

^§The SalI site for cloning is underlined.

^¶Lower case indicates the sequence in EcTet (31–252) that was removed after TEV proteolysis.

^††The underlined N-­terminal MA and C-terminal His₆-tag residues were added to the native sequence during cloning.