Part:BBa_K2550000:Design

T7 promoter Toehold Ribosome Switch with LacZ expression Lambert iGEM obtained a LacZ toehold construct assembled with a T7 promoter from the Styczynski Lab at Georgia Institute of Technology. The Toehold sequence used in the LacZ construct was obtained from the 144 first generation orthogonal toehold switches collection from the 2017 Collins paper titled Toehold Switches: De-Novo-Designed Regulators of Gene Expression. Following this unique toehold sequence is the LacZ operon. We introduced a base pair wobble in the LacZ gene that substituted an Adenine for a Guanine. The wobble mutation sequence was obtained from the Styczynski Lab at the Georgia Institute of Technology and was used to eliminate the illegal EcoRI site in the LacZ operon.