Parts

Parts for the Cell-Free System

Registry Number	Code used in Lab	Brief Description	Key Details	Biobricks used	Other sequence sources
BBa_K2450101	C100	mCherry - TEV protease (non-cleaving)	Doesn’t self-cleave	BBa_J06504; BBa_K1319004	Linker is Nd2 from https://www.hindawi.com/journals/bmri/2009/591923/tab2/
BBa_K2450201	C200	Modified TetR with TEV cleavage site - CFP	TEV cleavage site in linker between DNA binding and dimerisation domains	BBa_K106669	TEV cleavage site from Merck; CFP from AddGene
BBa_K2450251	C250	TetR - CFP	Normal TetR Class B	BBa_K106669	CFP from AddGene
BBa_K2450301	C300	Tet promoter - RBS - YFP	DNA part, contains promoter and RBS to give signal of YFP	BBa_R0040; BBa_B0030	eYFP from AddGene

Registry Number	Code used in Lab	Brief Description	Key Details	Biobricks used	Other sequence sources
BBa_K2450401	V100	N-SpyCatcher - OmpA - sfGFP	SpyCatcher is between leader sequence and OmpA. Used sfGFP to help it fold out of cytoplasm	n/a	Alves et al, 2016 AddGene
BBa_K2450451	v150	OmpA - C-SpyCatcher - sfGFP	SpyCatcher is after OmpA - will be compared to BBa_K2450401. Used sfGFP to help it fold out of cytoplasm	n/a	Alves et al, 2016
BBa_K2450501	v200	SpyTag - sfGFP - Quencher	TEV cleavage site between sfGFP and the quenching peptide	BBa_K1319014	Alves et al, 2016
BBa_K2450502	V200noQ	SpyTag - sfGFP	Control to check localisation to OMVs	n/a	Alves et al, 2016
BBa_K2450701	V500	sfGFP - N-terminal split TEV Protease - Inhibited Coiled Coil	Inhibited coiled coil contains a TEV cleavage site between the two helicies, leaving the acid coiled coil attached to the N-terminal split protease	n/a	Alves et al, 2016 Split TEV paper
BBa_K2450702	V600	mCherry - Inhibited Coiled Coil - C-terminal Split TEV Protease	Inhibited coiled coil contains a TEV protease cleavage site. This will result in the cleavage of the mCherry - Acid Coil being cleaved from the Base Coil - C Terminal Split TEV Protease.	n/a	Alves et al, 2016 Split TEV paper