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Summary: A dual selection module for directed evolution of genetic
circuits
YOHEI YOKOBAYASHIà
and FRANCES H. ARNOLD*
Division of Chemistry and Chemical Engineering, California Institute of Technology,
mail code 210-41, Pasadena, California 91125, USA; (*Author for correspondence,
e-mail: frances@cheme.caltech.edu); à
Present address: Department of Biomedical
Engineering, University of California, Davis, One Shields Avenue, Davis, CA 95616,
USA
Abstract. We have developed a genetic selection module which enables us to select for
positive and negative expression of a gene by controlling the expression of two antibiotic
resistance genes. This module can be used to tune the performance of genetic circuits
that yield simple ON/OFF gene expression outputs depending on the environment
(input). We have demonstrated the utility of the module by selecting a genetic inverter
from a 200 fold excess of nonfunctional inverters in two rounds.
Key words: cellular computation, directed evolution, genetic circuits, selection, syn-
thetic biology, synthetic gene circuits
Abbreviations: BLIP b - lactamase inhibitory protein; Cb carbenicillin; IPTG
isopropyl-b-D-thiogalactopyranoside; LB Luria-Bertani; Tc tetracycline
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