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:
Aug. 11, 2015

Filed:

Oct. 10, 2012
Applicant:

Dna Twopointo, Inc., Menlo Park, CA (US);

Inventors:

Jon E. Ness, Redwood City, CA (US);

Jeremy S. Minshull, Los Altos, CA (US);

Assignee:

DNA Twopointo, Inc., Menlo Park, CA (US);

Attorney:
Primary Examiner:
Int. Cl.
CPC ...
C12P 19/34 (2006.01); C12Q 1/68 (2006.01); C12N 15/66 (2006.01); C12N 9/90 (2006.01); C12N 15/10 (2006.01); C12N 15/64 (2006.01);
U.S. Cl.
CPC ...
C12N 15/66 (2013.01); C12N 9/90 (2013.01); C12N 15/10 (2013.01); C12N 15/64 (2013.01);
Abstract

Provided are methods, compositions, and kits for cloning of DNA using DNA topoisomerase. The methods comprise (I) combining into a mixture (A) a first polynucleotide comprising an origin of replication, a selectable marker, two topoisomerase recognition sequences, and two nicking agent recognition sequences, each of the topoisomerase recognition sequences being within 50 nucleotides of at least one of the nicking agent recognition sequences and each of the two nicking agent recognition sequences being nicked, with (B) a sequence-specific topoisomerase and (C) a second polynucleotide having 5' hydroxyl on each end; and (II) transforming the mixture into a host organism, thereby cloning the second polynucleotide. Formation or purification of a DNA-protein adduct prior to the addition of the second polynucleotide is not required. Also provided are vector sequences to facilitate performance of the methods and methods for modifying a vector of interest to render it useful in the disclosed methods.


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