Patent ID: 9115361
Filing Date: 2015-08-25
Classification: C07H,C12N

Abstract:
1. A method for simultaneously synthesizing an array of transgenes, comprising: a. providing a primary cloning vector plasmid comprising a first and a second docking point, b. introducing at least one Promoter nucleotide sequence into a corresponding Promoter shuttle vector; c. introducing at least one Expression nucleotide sequence into a corresponding Expression shuttle vector; d. introducing at least one Regulatory nucleotide sequence into a corresponding Regulatory shuttle vector; e. simultaneously ligating the Promoter, Expression, and Regulatory nucleotide sequences from the Promoter, Expression, and Regulatory shuttle vectors to the cloning vector plasmid, between the first and second docking points, thereby forming a transgene construct comprising the Promoter, Expression, and Regulatory nucleotide sequences; f. digesting the cloning vector plasmid with homing endonucleases that recognize the unique homing endonuclease sites in (a), thereby releasing the transgene construct; g. providing a second cloning vector plasmid comprising the same unique homing endonuclease site in a forward orientation and the same unique homing endonuclease site in a reverse orientation as the first cloning vector plasmid; h. digesting the second cloning vector plasmid with the homing endonuclease used in (f); and i. ligating the transgene construct of (f) into the second cloning vector plasmid.