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. 07, 2025

Filed:

Dec. 17, 2021
Applicant:

New England Biolabs, Inc., Ipswich, MA (US);

Inventors:

Gregory Lohman, Cambridge, MA (US);

Vladimir Potapov, Auburndale, MA (US);

John M. Pryor, Gerogetown, MA (US);

Rebecca Kucera, Hamilton, MA (US);

Katharina Bilotti, Burlington, MA (US);

Richard D. Morgan, Middleton, MA (US);

Assignee:

New England Biolabs, Inc., Ipswich, MA (US);

Attorney:
Primary Examiner:
Int. Cl.
CPC ...
C12N 15/10 (2006.01); C12P 19/34 (2006.01); C12Q 1/6855 (2018.01);
U.S. Cl.
CPC ...
C12N 15/1068 (2013.01); C12P 19/34 (2013.01); C12Q 1/6855 (2013.01);
Abstract

Ordered assembly of large numbers of fragments into a single large DNA have been improved in both frequency and fidelity of the assembled product. This has been achieved by novel compositions and methods that are utilized in a computer system that integrates comprehensive ligation data from multiple sources to provide optimized synthetic overhangs or overhangs from restriction endonuclease cleavage on DNA fragments for assembly by ligation. Intragenic cut sites are avoided by the use of a novel restriction endonuclease which recognizes 7 nucleotides (bases) and cuts DNA to create 4-base overhangs with the help of a synthetic activator oligonucleotide. Variations in ligation preferences by different ligases provide extra precision in assembly reactions. The use of the improved methods are exemplified by the successful assembly from 52 fragments of a viral genome and also a 52 fragment ordered assembly of a bacteria operon.


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