	US 12,404,524 B2
	Expression vector
Robert Joseph Stankey, Madison, WI (US); and David Mead, Middleton, WI (US)
Assigned to TERRA BIOWORKS, INC., Middleton, WI (US)
Filed by TERRA BIOWORKS, INC., Middleton, WI (US)
Filed on Jul. 18, 2022, as Appl. No. 17/813,088.
Application 17/813,088 is a continuation of application No. 16/815,399, filed on Mar. 11, 2020, granted, now 11,390,882.
Claims priority of provisional application 62/817,345, filed on Mar. 12, 2019.
Prior Publication US 2022/0372517 A1, Nov. 24, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/52 (2006.01); C12N 9/00 (2006.01); C12N 15/09 (2006.01); C12N 15/70 (2006.01); C12N 15/86 (2006.01); C12Q 1/68 (2018.01); C40B 40/08 (2006.01)

CPC C12N 15/86 (2013.01) [C12N 15/52 (2013.01); C12N 15/70 (2013.01)]

16 Claims

1. A method of expressing a polypeptide from an insert nucleic acid comprising a biosynthetic gene cluster, the method comprising:

providing a nucleic acid comprising a biosynthetic gene cluster;

providing an expression vector comprising a first promoter and a second promoter flanking a cloning site, wherein the first promoter and the second promoter direct transcription toward each other and in opposite directions;

cloning an insert nucleic acid comprising the biosynthetic gene cluster at said cloning site to provide a recombinant expression vector comprising said insert nucleic acid;

transforming a host cell with the recombinant expression vector;

expressing a polypeptide from the biosynthetic gene cluster; and

detecting the expression of the polypeptide using a functional selection or screen that is specific for the polypeptide or for a product that is indicative of the polypeptide.