Production of vaccinia capping enzyme
AssigneeGinkgo Bioworks, Inc.
InventorsJosef Bober, Jeffrey Ian Boucher, Justin Michael Gardin, Jason King, Scott Marr, Matthew McMahon, Krishnaben S. Patel, Abraham Waldman
Life of this patent
Estimating the term…
Abstract
Aspects of the disclosure relate to production of vaccinia capping enzyme (VCE) in host cells. For example, host cells may comprise: a promoter; a ribosome binding site (RBS); a nucleic acid encoding a vaccinia capping enzyme (VCE) or VCE subunit; and a terminator.
Claims44
A non-naturally occurring nucleic acid comprising:
- a) a promoter, wherein the promoter comprises a sequence that is at least 90% identical to SEQ ID NO:
- 8 or 9;
- and
- b) a nucleic acid encoding an amino acid sequence that is at least 90% identical to SEQ ID NO:
- 6 or 29, and/or a nucleic acid encoding an amino acid sequence that is at least 90% identical to SEQ ID NO:
- 7 or 31,
- wherein (a) and (b) are operably linked, and wherein the non-naturally occurring nucleic acid further comprises a ribosome binding site (RBS).
A non-naturally occurring nucleic acid comprising:
- a) a first promoter, wherein the first promoter comprises a sequence that is at least 90% identical to SEQ ID NO:
- 8 or 9;
- b) a first nucleic acid, wherein the first nucleic acid encodes an amino acid sequence that is at least 90% identical to SEQ ID NO:
- 6 or 29;
- c) a second promoter, wherein the second promoter comprises a sequence that is at least 90% identical to SEQ ID NO:
- 8 or 9;
- and
- d) a second nucleic acid, wherein the second nucleic acid encodes an amino acid sequence that is at least 90% identical to SEQ ID NO:
- 7 or 31,
- wherein (a) and (b) are operably linked, and wherein (c) and (d) are operably linked, and wherein the non-naturally occurring nucleic acid further comprises at least one ribosome binding site (RBS).
A non-naturally occurring nucleic acid comprising a sequence that is at least 90% identical to any one of SEQ ID NOs:
- 21-28, or 49-54.
A host cell comprising one or more non-naturally occurring nucleic acids comprising:
- a promoter, wherein the promoter comprises a sequence that is at least 90% identical to SEQ ID NO:
- 8 or 9, and
- a nucleic acid encoding an amino acid sequence that is at least 90% identical to SEQ ID NO:
- 6 or 29 and/or a nucleic acid encoding an amino acid sequence that is at least 90% identical to SEQ ID NO:
- 7 or 31,
- wherein one or more of the non-naturally occurring nucleic acids further comprise a ribosome binding site (RBS).
A host cell comprising one or more non-naturally occurring nucleic acids comprising:
- a) a first promoter, wherein the first promoter comprises a sequence that is at least 90% identical to SEQ ID NO:
- 8 or 9;
- b) a first nucleic acid, wherein the first nucleic acid encodes an amino acid sequence that is at least 90% identical to SEQ ID NO:
- 6 or 29;
- c) a second promoter, wherein the second promoter comprises a sequence that is at least 90% identical to SEQ ID NO:
- 8 or 9;
- and
- d) a second nucleic acid, wherein the second nucleic acid encodes an amino acid sequence that is at least 90% identical to SEQ ID NO:
- 7 or 31,
- wherein (a) and (b) are operably linked, and wherein (c) and (d) are operably linked, and wherein one or more of the non-naturally occurring nucleic acids further comprises at least one ribosome binding site (RBS).
A non-naturally occurring nucleic acid comprising:
- (a) a promoter, wherein the promoter is a Ptac promoter or a functional fragment thereof, or a P(T5) 2xlacO promoter or a functional fragment thereof;
- and
- (b) a nucleic acid encoding a D1 subunit of VCE and/or a D12 subunit of vaccinia capping enzyme,
- wherein (a) and (b) are operably linked, and wherein the non-naturally occurring nucleic acid further comprises a ribosome binding site (RBS).
Dependencies are inferred from the claim text.