Escherichia coli strain:NEB10beta Raw sequence reads

We built a synthetic system in Escherichia coli to study how antisense transcription can control the expression level of a gene and tune the response characteristics of a regulatory circuit. We develop a genetic part that consists of a unidirectional terminator followed by a constitutive antisense promoter and demonstrate that repression of gene expression is proportional to the antisense promoter strength. Chip-based oligo synthesis is applied to build a large library of 5668 terminator-promoter combinations that is used to control the expression of three repressors (PhlF, SrpR, and TarA) in a simple genetic circuit (NOT gate). The library sorted using FACS and deep sequenced to demonstrate that antisense promoters can be used to tune the threshold of the response function without impacting other properties.