The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.

The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.

Date of Patent:
Jan. 03, 2017

Filed:

Mar. 30, 2010
Applicants:

Lei Young, Gaithersburg, MD (US);

Hamilton O. Smith, San Diego, CA (US);

Daniel Glenn Gibson, Crofton, MD (US);

Inventors:

Lei Young, Gaithersburg, MD (US);

Hamilton O. Smith, San Diego, CA (US);

Daniel Glenn Gibson, Crofton, MD (US);

Assignee:

Synthetic Genomics, Inc., La Jolla, CA (US);

Attorney:
Primary Examiner:
Int. Cl.
CPC ...
C12N 9/00 (2006.01); C12Q 1/68 (2006.01); C12N 15/66 (2006.01); C12N 15/10 (2006.01); C12N 15/64 (2006.01); C12P 19/34 (2006.01);
U.S. Cl.
CPC ...
C12Q 1/6862 (2013.01); C12N 15/10 (2013.01); C12N 15/64 (2013.01); C12N 15/66 (2013.01); C12P 19/34 (2013.01);
Abstract

The present invention relates, e.g., to in vitro method, using isolated protein reagents, for joining two double stranded (ds) DNA molecules of interest, wherein the distal region of the first DNA molecule and the proximal region of the second DNA molecule share a region of sequence identity, comprising contacting the two DNA molecules in a reaction mixture with (a) a non-processive 5' exonuclease; (b) a single stranded DNA binding protein (SSB) which accelerates nucleic acid annealing; (c) a non strand-displacing DNA polymerase; and (d) a ligase, under conditions effective to join the two DNA molecules to form an intact double stranded DNA molecule, in which a single copy of the region of sequence identity is retained. The method allows the joining of a number of DNA fragments, in a predetermined order and orientation, without the use of restriction enzymes.


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