Characterization of 582 natural and synthetic terminators and quantification of their design constraints.

Chen YJ, Liu P, Nielsen AA, Brophy JA, Clancy K, Peterson T, Voigt CA
Nat Methods. 2013 Jun 2. doi: 10.1038/nmeth.2515. PubMed Article

By studying a collection of 582 natural and synthetic terminators, Christopher Voigt’s lab has characterized the strength of various terminators and identified design constraints required to build strong, reliable synthetic terminators. This expands the library of terminators available for use in synthetic biology circuits.

This publication describes the measurements made on 227 E. coli terminators, 90 reverse terminators, and 265 synthetic terminators. The terminator strength was measured by developing assay reporter plasmids (shown below), where the terminator of interest was inserted between the GFP and RFP, and comparing the induced GFP and induced RFP output. Using these measurements, Chen et al. modeled the contribution of the various features (U-tract, A-tract, hairpin, etc.) on terminator strength and designed new synthetic terminators.

The Voigt lab has deposited 10 of the described plasmids to Addgene. These include the empty backbone, pGR, and 9 variations of pGR-** with various terminators inserted between the fluorescent proteins (where ** is the specific terminator). The sequences and corresponding data for all 582 terminators can be found below.

Resources

Downloadable data and sequences for all 582 terminators:

• Excel Sheet - Natural Terminators 126.8 KB

• Excel Sheet - Synthetic Terminators 94.7 KB

• Excel Sheet - Removed Terminators 18.1 KB

Plasmids from Article

ID	Plasmid	Purpose
46002	pGR	Reporter plasmid for measuring terminator strength by inserting terminator of interest between the GFP and RFP, expression driven by a single PBAD promoter inducible by arabinose
46003	pGR-BBa_B0010
46004	pGR-L3S2P21
46005	pGR-L3S2P00
46006	pGR-ECK120015170
46007	pGR-L3S2P36
46008	pGR-ECK125108723
46009	pGR-ECK120010783
46010	pGR-ECK120010800
46011	pGR-ECK120033127