US 12,338,470 B2
Enzymes and systems for synthesizing DNA
Paul F. Predki, Carlsbad, CA (US); and Stefen Boehme, Encinitas, CA (US)
Assigned to IRIDIA, INC., Carlsbad, CA (US)
Filed by IRIDIA, INC., Carlsbad, CA (US)
Filed on Apr. 11, 2023, as Appl. No. 18/298,995.
Application 18/298,995 is a division of application No. 16/866,439, filed on May 4, 2020, granted, now 11,655,465.
Claims priority of provisional application 62/842,333, filed on May 2, 2019.
Prior Publication US 2024/0084284 A1, Mar. 14, 2024
Int. Cl. C12N 9/90 (2006.01); B01L 3/00 (2006.01); C12N 15/74 (2006.01); C12P 19/34 (2006.01)
CPC C12N 9/90 (2013.01) [B01L 3/508 (2013.01); C12N 15/74 (2013.01); C12P 19/34 (2013.01); C12Y 599/01002 (2013.01); B01L 2300/047 (2013.01)] 5 Claims
 
1. A topoisomerase comprising the following sequence:
SEQ ID NO: 3
RALFYKDGKLFTDNNFLNPVSDDNPAYEVLQHVKIPTHLTDVVVYEQTWE
 
EALTRLIFVGSDSKGX1RX2X3FYGKMHVQNX4NAKRDRIFVRVYNVMKR
 
INCFINKNIKKSSTDSNYQLAVFMLMETMFFIRFGKMKYLKENETVGLLT
 
LKNKHIEISPDEIVIKFVGKDKVSHEFVVHKSNRLYKPLLKLTDDSSPEE
 
FLFNKLSERKVYECIKQFGIRIKDLRTYGVNYTFLYNFWTNVKSISPLPS
 
PKKLIALTIKQTAEVVGHTPSISKRAYMATTILEMVKDKNFLDVVSKTTF
 
DEFLSIVVDHVKSSTDG
wherein at least two of X1, X2, X3, and X4, are mutated from the residues in the wild type sequence [wherein in the wild-type sequence, X1 is arginine (R), X2 is glutamine (Q), X3 is tyrosine (Y), and X4 is arginine (R)] to glycine (G), alanine (A) or valine (V);
provided that where only two residues are mutated, X1 and X2 are not both mutated to A.