BioFab Terminator

his [min]

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Description

Part of the Biofab collection of terminators. These are natural and synthetic hairpin terminators that were characterized and modelled to insulate upstream regions from downstream polymerase (RNAP) flow. Termination is performed by the detachment of RNAP from DNA due to the highly stable hairpin structure and the following U-rich tail.

This collection includes many other terminator sequences, but this stood out for having > 95% termination strength.

Characterization

Parts were characterized on E. coli BW25113 in rich medium, in 96-well flow citometer assays. Termination efficiency was calculated by measuring the fluorescence from reporters (RFP and GFP) coded both upstream and downstream of the terminator, and calculating the ratio of read-through insulated (1 - upstream/downstream fluorescence). This fluorescence was measured using flow citometry with automated gating.
RNAse sites flanking the terminator were used to detach the coding sequences from each other and the terminator.

Sequences

Terminator

GCCCCCGGAAGATCACCTTCCGGGGGCTTTTTTATT

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FASTA

Reference

Cambray, G. et al. Measurement and modeling of intrinsic transcription terminators. Nucleic Acids Res. 41, 5139–5148 (2013).