Abstract: The present disclosure is generally related to novel promoter sequences and methods thereof for enhanced protein production in Bacillus sp. (host) cells. As set forth herein, the novel promoter sequences of the disclosure, when operably linked to a gene or open reading frame encoding a protein of interest, are particularly well suited for use in large scale production of industrially relevant proteins.

Abstract: Methods and compositions are provided for modifying the genome of Bacillus sp. cells without the use of a selectable marker and without the use of a guided Cas endonuclease. The disclosure includes methods for integrating donor DNA sequences into the genome of a Bacillus sp. cell without the use of a selectable marker and without the use of Cas endonucleases into said genome, as well as methods for deleting genes of interest and/or providing point mutations into the genome of Bacillus sp. cells.

Abstract: Methods and compositions are provided for integrating donor DNA sequences into the genome of a Bacillus sp. cell without the integration of a selectable marker into said genome. The methods employ a linear recombinant DNA construct comprising a donor DNA flanked by long homology arms (each of at least 1000 nucleotides in length) in combination with a recombinant DNA construct encoding a Cas9 endonuclease and a guide RNA, for the introduction of a guide RNA/Cas endonuclease into a Bacillus sp. cell, and as such providing a highly effective system for integrating donor DNA sequences into the genome of said Bacillus sp. cell, without the need to integrate a selectable marker in the genome of said Bacillus sp. cell.

Abstract: Methods and compositions are provided for integrating genes of interest into the genome of a Bacillus sp. cell without the integration of a selectable marker into said genome. The methods employ a dual circular recombinant DNA system for introduction of a guide RNA/Cas endonuclease system (also referred to as an RNA guided endonuclease, RGEN) as well as a donor DNA into a Bacillus sp. cell, and providing a highly effective system for inserting genes of interest into the genome of said Bacillus sp. cell.

Abstract: The present disclosure is generally related to novel promoter sequences and methods thereof for enhanced protein production in Bacillus sp. (host) cells. As set forth herein, the novel promoter sequences of the disclosure, when operably linked to a gene or open reading frame encoding a protein of interest, are particularly well suited for use in large scale production of industrially relevant proteins.