SEQUENCE FOR PROTEIN DECAY

§102 §103 §112

Notice of Pre-AIA or AIA Status The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA . DETAILED ACTION This application is 371 of PCT/EP2022/081589. The amendment filed on August 19, 2026 has been entered. Status of Claims Claims 1-26 are pending. Claims 1-26 are under examination. Response to Amendments/Arguments Nucleotide and/or Amino Acid Sequence Disclosures Applicant’s arguments, see page 8 of the Remarks, filed August 19, 2026, with respect to the specification have been fully considered and are persuasive. The specification has been amended to insert sequence identifier for amino acid sequence at page 2. Therefore, the objection to the specification has been withdrawn. Claim Rejections - 35 USC § 112(b) The following is a quotation of 35 U.S.C. 112(b): (b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention. The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph: The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention. Withdrawn Rejections Applicant’s arguments, see pages 9-10 of the Remarks, filed August 19, 2026, with respect to claim 3 have been fully considered and are persuasive. Claim 3 has been amended to clarify the nucleotide sequence(s) of d). Therefore, the rejection of claim 3 under 35 U.S.C. 112(b) has been withdrawn. Applicant’s arguments, see page 11 of the Remarks, filed August 19, 2026, with respect to claims 7, 9-10 and 13 have been fully considered and are persuasive. Claims 7, 9-10 and 13 have been amended to delete the indefinite phrase “preferably”. Therefore, the rejection of claims 7, 9-10 and 13 under 35 U.S.C. 112(b) has been withdrawn. Maintained Rejection Claim 5 is rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention. The term “gradually” in claim 5 is a relative term which renders the claim indefinite. The term “minimal” is not defined by the claim, the specification does not provide a standard for ascertaining the requisite degree, and one of ordinary skill in the art would not be reasonably apprised of the scope of the invention. It is unclear as to what degree of reduction in the level of the fusion protein is considered as “gradual”. A perusal of the specification did not provide a clear definition for the above limitation. Without a clear definition in terms of numerical value, those skilled in the art would be unable to ascertain what degree of reduction in the level of the fusion protein is gradual. Applicant's arguments filed August 19, 2026 have been fully considered but they are not persuasive. Applicant argues that since the term “gradually” is explicitly defined by the application at paragraph [0095], term is definite. This is not found persuasive. Although the claims are interpreted in light of the specification, limitations from the specification are not read into the claims. See In re Van Geuns, 988 F.2d 1181, 26 USPQ2d 1057 (Fed. Cir. 1993). “Though understanding the claim language may be aided by explanations contained in the written description, it is important not to import into a claim limitations that are not part of the claim. For example, a particular embodiment appearing in the written description may not be read into a claim when the claim language is broader than the embodiment….(“We have cautioned against reading limitations into a claim from the preferred embodiment described in the specification, even if it is the only embodiment described, absent clear disclaimer in the specification.”).”, see MPEP 2111.I. and MPEP 2111.II. Therefore, the rejection has been maintained. New Rejection Claim 1 and claims 2 and 5-26 depending therefrom is rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor, or for pre-AIA the applicant regards as the invention. Claim 1 recites the phrase “the fragment is at least 18 contiguous amino acids long”. The metes and bounds of the phrase in the context of the claim are not clear. The recitation of “contiguous” is confusing and redundant in defining the fragment because a fragment is comprised of contiguous amino acids. Appropriate correction is required. The following is a quotation of the first paragraph of 35 U.S.C. 112(a): (a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention. The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112: The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention. Claims 1-3 and 5-26 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for pre-AIA the inventor(s), at the time the application was filed, had possession of the claimed invention. MPEP 2111.01 states that ''[d]uring examination, the claims must be interpreted as broadly as their terms reasonably allow.'' The limitation “an amino acid sequence” recited in claim 1 has been broadly interpreted as any amino acid sequence comprising as little as two contiguous amino acids of SEQ ID NO:1. The limitation “fragment thereof…wherein the fragment is at least 18 contiguous amino acids long” has been broadly interpreted as any amino acid sequence comprising any two contiguous amino acids of SEQ ID NO:1 and has a length of at least 18 contiguous amino acids. Therefore, the claims are directed to an expression cassette, vector, or host cell comprising a polynucleotide encoding a fusion protein comprising (a) any polypeptide comprising as little as two contiguous amino acids of SEQ ID NO:1, any polypeptide comprising as little as two contiguous amino acids of SEQ ID NO:1 and having a length of 18 contiguous amino acids, or any polypeptide having at least 70% sequence identity to SEQ ID NO:1, wherein the polypeptide has the function directing protein decay and (b) a protein of interest. Thus, the claims are directed to an expression cassette, vector or host cell comprising a fusion protein comprising (a) a genus of polypeptides having unknown structure, wherein the polypeptide has the function directing protein decay, and (b) a protein of interest. MPEP 2163 I. states that to “satisfy the written description requirement, a patent specification must describe the claimed invention in sufficient detail that one skilled in the art can reasonably conclude that the inventor had possession of the claimed invention. MPEP 2163. II.A.3.(a) sates that “Possession may be shown in many ways. For example, possession may be shown by describing an actual reduction to practice of the claimed invention. Possession may also be shown by a clear depiction of the invention in detailed drawings or in structural chemical formulas which permit a person skilled in the art to clearly recognize that inventor had possession of the claimed invention. An adequate written description of the invention may be shown by any description of sufficient, relevant, identifying characteristics so long as a person skilled in the art would recognize that the inventor had possession of the claimed invention. According to MPEP 2163.II.A.3.(a).ii), “Satisfactory disclosure of a ‘representative number’ depends on whether one of skill in the art would recognize that the applicant was in possession of the necessary common attributes or features possessed by the members of the genus in view of the species disclosed. For inventions in an unpredictable art, adequate written description of a genus which embraces widely variant species cannot be achieved by disclosing only one species within the genus…Instead, the disclosure must adequately reflect the structural diversity of the claimed genus, either through the disclosure of sufficient species that are ‘representative of the full variety or scope of the genus,’ or by the establishment of ‘a reasonable structure-function correlation.’" The recitation of “directs protein decay” fails to provide a sufficient description of the genus of the polypeptides as it merely describes the functional features of the genus without providing any definition of the structural features of the species within the genus. The specification does not specifically define any of the species that fall within the genus. The specification does not define any structural features commonly possessed by members of the genus that distinguish them from others. One skilled in the art therefore cannot, as one can do with a fully described genus, visualize or recognize the identity of the members of the genus. The polypeptide comprising the amino acid sequence of SEQ ID NO:1 was not known in the prior art. A search of SEQ ID NO:1 did not result in polypeptides having any significant similarity, as shown below. PNG media_image1.png 744 1194 media_image1.png Greyscale Fransceus (J Ind Microbiol Biotechnol. 2017 May;44(4-5):687-695. – cited previously on form PTO-892) reviews protein engineering techniques, such as random mutagenesis and recombination, directed evolution and iterative or combinatory saturation “hotspots”. Fransceus states that “a recurring problem, however, is choosing which amino acid positions should be targeted. Answering this question is not an easy feat and requires substantial insight in the relationship between an enzyme’s sequence or structure and its properties.” Sanavia (Computational and Structural Biotechnology Journal, Volume 18, 2020, Pages 1968-1979. – cited previously on form PTO-892) discloses challenges in the prediction of protein stability in the occurrence of multiple mutations. “Multiple-point mutations are common variations of the protein sequence that may be needed in protein engineering when a single-point mutation is not enough to yield the desired stability change. Dealing with multiple-site variations adds another level of complexity beyond the prediction of the effect of a single variant on protein stability, since it requires the learning of many types of combinatorial effects”. The specification is limited to description of an expression cassette, vector, or host cell comprising a polynucleotide encoding a fusion protein comprising (a) the polypeptide comprising the amino acid sequence of SEQ ID NO:1 and (b) a protein of interest, wherein the polypeptide of (a) directs decay of the protein of interest of (b). While MPEP 2163 acknowledges that in certain situations “one species adequately supports a genus,” it also acknowledges that “[f]or inventions in an unpredictable art, adequate written description of a genus which embraces widely variant species cannot be achieved by disclosing only one species within the genus.” In view of the widely variant species encompassed by the genus, the example described above is not enough and does not constitute a representative number of species to describe the whole genus. Therefore, the specification fails to describe a representative species of the claimed genus. Further, one of skill in the art could identify fragments and polypeptides having at least 70% sequence identity to SEQ ID NO:1. However, there is no teaching regarding which amino acid or which 30% of the amino acids can vary from SEQ ID NO:1 and result in polypeptide having the function of directing protein decay. An important consideration is that structure is not necessarily a reliable indicator of function. In the instant case, there is no disclosure relating similarity of structure to conservation of function. Conservation of structure is not necessarily a surrogate for conservation of function. Given this lack of description of the representative species encompassed by the genus of the claims, the specification fails to sufficiently describe the claimed invention in such full, clear, concise, and exact terms that a skilled artisan would recognize that applicants were in possession of the inventions of claims 1-3 and 5-26. Applicant's arguments filed August 19, 2026 have been fully considered but they are not persuasive. Applicant argues that (A) the present application provides a clear and structurally define reference point in the form of the specific 31 amino acid sequence disclosed as SEQ ID NO:1, (B) paragraph [083] explicitly defines “decay sequence” as this sequence, (C) the application consistently defines the decay sequence as encompassing sequence having at least 60-95% sequence identity to SEQ ID NO:1, and (D) a sequence identity threshold of at least 70% in the context of a 31 amino acid sequence corresponds to a maximum of only 9 amino acid substitutions. This is not found persuasive. The claims are not limited to the decay sequence having the amino acid sequence of SEQ ID NO:1. (B)-(D) One of skill in the art could identify fragments and polypeptides having at least 70% sequence identity to SEQ ID NO:1. However, there is no teaching regarding which amino acid or which 30% of the amino acids can vary from SEQ ID NO:1 and result in polypeptide having the function of directing protein decay. An important consideration is that structure is not necessarily a reliable indicator of function. In the instant case, there is no disclosure relating similarity of structure to conservation of function. Conservation of structure is not necessarily a surrogate for conservation of function. Applicant argues that that skilled person would understand that the decay function is associated with the specific structural context of the disclosed sequence, namely a host peptide tag derived from a defined frameshift event and the small size of the sequence and its role as a degrading tag, minor sequences, would reasonably be expected to retain the functional property. This is not found persuasive. The polypeptide comprising the amino acid sequence of SEQ ID NO:1 was not known in the prior art. A search of SEQ ID NO:1 did not result in polypeptides having any significant similarity, as shown above. Fransceus (J Ind Microbiol Biotechnol. 2017 May;44(4-5):687-695. – cited previously on form PTO-892) reviews protein engineering techniques, such as random mutagenesis and recombination, directed evolution and iterative or combinatory saturation “hotspots”. Fransceus states that “a recurring problem, however, is choosing which amino acid positions should be targeted. Answering this question is not an easy feat and requires substantial insight in the relationship between an enzyme’s sequence or structure and its properties.” Sanavia (Computational and Structural Biotechnology Journal, Volume 18, 2020, Pages 1968-1979. – cited previously on form PTO-892) discloses challenges in the prediction of protein stability in the occurrence of multiple mutations. “Multiple-point mutations are common variations of the protein sequence that may be needed in protein engineering when a single-point mutation is not enough to yield the desired stability change. Dealing with multiple-site variations adds another level of complexity beyond the prediction of the effect of a single variant on protein stability, since it requires the learning of many types of combinatorial effects”. Applicant argues that (A) with respect to “fragments” of SEQ ID NO:1, such fragments are properly constructed in light of the description and limited to fragments at least 18 contiguous amino acids and (B) the interpretation of “fragment” as asserted by the Office Action is not based on the definition of such term in paragraph [0077] of the instant application and when properly constructed, the claimed subject matter is clearly delimited and does not give rise to the “essentially infinite” sequence alleged in the Office Action. This is not found persuasive. Paragraph [0077] recites “In a preferred embodiment..”. Although the claims are interpreted in light of the specification, limitations from the specification are not read into the claims. See In re Van Geuns, 988 F.2d 1181, 26 USPQ2d 1057 (Fed. Cir. 1993). “Though understanding the claim language may be aided by explanations contained in the written description, it is important not to import into a claim limitations that are not part of the claim. For example, a particular embodiment appearing in the written description may not be read into a claim when the claim language is broader than the embodiment….(“We have cautioned against reading limitations into a claim from the preferred embodiment described in the specification, even if it is the only embodiment described, absent clear disclaimer in the specification.”).”, see MPEP 2111.I. and MPEP 2111.II. In the instant case, claim 1 recites that the nucleotide sequence encodes (1) an amino acid sequence shown in SEQ ID NO:1 or (2) a fragment thereof… wherein the fragment is at least 18 contiguous amino acids long. The limitation “an amino acid sequence” recited in claim 1 has been broadly interpreted as any amino acid sequence comprising as little as two contiguous amino acids of SEQ ID NO:1. The limitation “fragment thereof…wherein the fragment is at least 18 contiguous amino acids long” has been broadly interpreted as any amino acid sequence comprising any two contiguous amino acids of SEQ ID NO:1 and has a length of at least 18 contiguous amino acids. Therefore, the claims are directed to an expression cassette, vector or host cell comprising a fusion protein comprising (a) a genus of polypeptides having unknown structure, wherein the polypeptide has the function directing protein decay, and (b) a protein of interest. Applicant argues that the claimed fragments are fully described because the application provides a clear and complete disclosure of the full-length decay sequence (SEQ ID NO:1), the application consistently teaches this sequence function as a decay sequence, and the claimed fragments are contiguous and identical portions of this explicitly disclosed sequence. This is not found persuasive. As discussed above, the claims are not limited to the decay sequence having the amino acid sequence of SEQ ID NO:1. One of skill in the art could identify fragments and polypeptides having at least 70% sequence identity to SEQ ID NO:1. However, there is no teaching regarding which amino acid or which 30% of the amino acids can vary from SEQ ID NO:1 and result in polypeptide having the function of directing protein decay. An important consideration is that structure is not necessarily a reliable indicator of function. In the instant case, there is no disclosure relating similarity of structure to conservation of function. Conservation of structure is not necessarily a surrogate for conservation of function. Applicant argues that Applicant has found a novel amino acid sequence and “the 31 amino acid sequence (SEQ ID NO:1) resulting from the frameshift in the 3’-region of ERG7 close to the wildtype stop codon seem to direct protein decay” ([0034]-[0035]) and the present application therefore provides a clear and sufficient description of the genus of strains utilized and defines both the functional and structural reference point in the for of the novel specific 31 amino acid sequence disclosed as SEQ ID NO:1. This is not found persuasive. As discussed above, one of skill in the art could identify fragments and polypeptides having at least 70% sequence identity to SEQ ID NO:1. However, there is no teaching regarding which amino acid or which 30% of the amino acids can vary from SEQ ID NO:1 and result in polypeptide having the function of directing protein decay. An important consideration is that structure is not necessarily a reliable indicator of function. In the instant case, there is no disclosure relating similarity of structure to conservation of function. Conservation of structure is not necessarily a surrogate for conservation of function. Applicant argues that since the inventors rebuild and found a novel amino acid sequence (SEQ ID NO:1), a search of SEQ ID NO:1 as performed on NCBI Blast search would not yield the SEQ ID NO:1 as described in the present invention. Applicant argues that since the application as filed further describes the “decay sequence”, which refer to the amino acid sequence shown in SEQ DI NO:1 or fragments thereof, the claims are fully described. This is not found persuasive. The polypeptide comprising the amino acid sequence of SEQ ID NO:1 was not known in the prior art and therefore, a search of SEQ ID NO:1 in a NCBI Blast search does not yield an amino acid sequence having 100% sequence identity to SEQ ID NO:1. However, a search of SEQ ID NO:1 did not result in polypeptides having any significant similarity, as shown above. There is no teaching regarding which amino acid or which 30% of the amino acids can vary from SEQ ID NO:1 and result in polypeptide having the function of directing protein decay. An important consideration is that structure is not necessarily a reliable indicator of function. In the instant case, there is no disclosure relating similarity of structure to conservation of function. Conservation of structure is not necessarily a surrogate for conservation of function. Fransceus (J Ind Microbiol Biotechnol. 2017 May;44(4-5):687-695. – cited previously on form PTO-892) reviews protein engineering techniques, such as random mutagenesis and recombination, directed evolution and iterative or combinatory saturation “hotspots”. Fransceus states that “a recurring problem, however, is choosing which amino acid positions should be targeted. Answering this question is not an easy feat and requires substantial insight in the relationship between an enzyme’s sequence or structure and its properties.” Sanavia (Computational and Structural Biotechnology Journal, Volume 18, 2020, Pages 1968-1979. – cited previously on form PTO-892) discloses challenges in the prediction of protein stability in the occurrence of multiple mutations. “Multiple-point mutations are common variations of the protein sequence that may be needed in protein engineering when a single-point mutation is not enough to yield the desired stability change. Dealing with multiple-site variations adds another level of complexity beyond the prediction of the effect of a single variant on protein stability, since it requires the learning of many types of combinatorial effects”. Therefore, the rejection has been maintained. Claims 1-3 and 5-26 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, because the specification, while being enabling for an expression cassette, vector, or host cell comprising a polynucleotide encoding a fusion protein comprising (a) the polypeptide comprising the amino acid sequence of SEQ ID NO:1 and (b) a protein of interest, wherein the polypeptide of (a) directs decay of the protein of interest of (b), does not reasonably provide enablement an expression cassette, vector, or host cell comprising a polynucleotide encoding a fusion protein comprising (a) any polypeptide having unknown structure and having the function of directing protein decay and (b) a protein of interest. The specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the invention commensurate in scope with these claims. Factors to be considered in determining whether undue experimentation is required are summarized in In re Wands 858 F.2d 731, 8 USPQ2nd 1400 (Fed. Cir, 1988). They include (1) the quantity of experimentation necessary, (2) the amount of direction or guidance presented, (3) the presence or absence of working examples, (4) the nature of the invention, (5) the state of the prior art, (6) the relative skill of those in the art, (7) the predictability or unpredictability of the art, and (8) the breadth of the claims. The breadth of the claims. MPEP 2111.01 states that ''[d]uring examination, the claims must be interpreted as broadly as their terms reasonably allow.'' The limitation “an amino acid sequence” recited in claim 1 has been broadly interpreted as any amino acid sequence comprising as little as two contiguous amino acids of SEQ ID NO:1. The limitation “fragment thereof…wherein the fragment is at least 18 contiguous amino acids long” has been broadly interpreted as any amino acid sequence comprising any two contiguous amino acids of SEQ ID NO:1 and has a length of at least 18 contiguous amino acids. Therefore, the claims are directed to an expression cassette, vector, or host cell comprising a polynucleotide encoding a fusion protein comprising (a) any polypeptide comprising as little as two contiguous amino acids of SEQ ID NO:1, any polypeptide comprising as little as two contiguous amino acids of SEQ ID NO:1 and having a length of 18 contiguous amino acids, or any polypeptide having at least 70% sequence identity to SEQ ID NO:1, wherein the polypeptide has the function directing protein decay and (b) a protein of interest. Thus, the claims are directed to an expression cassette, vector or host cell comprising a fusion protein comprising (a) a polypeptide having unknown structure, wherein the polypeptide has the function of directing protein decay and (b) a protein of interest. The claims are not commensurate with the enablement provided by the disclosure with regard to the extremely large number of polypeptides having the function of directing protein decay. In the instant case, the specification is limited to an expression cassette, vector, or host cell comprising a polynucleotide encoding a fusion protein comprising (a) the polypeptide comprising the amino acid sequence of SEQ ID NO:1 and (b) a protein of interest, wherein the polypeptide of (a) directs decay of the protein of interest of (b). The quantity of experimentation required to practice the claimed invention based on the teachings of the specification. While protein isolation techniques, recombinant and mutagenesis techniques were known in the art at the time of the invention, e.g. mutagenesis, and it is routine in the art to screen for variants comprising multiple substitutions or multiple modifications as encompassed by the instant claims, the specific amino acid positions within the protein's sequence where amino acid modifications can be made with a reasonable expectation of success in obtaining the desired activity/utility are limited in any protein and the result of such modifications is unpredictable. In addition, one skilled in the art would expect any tolerance to modification for a given protein to diminish with each further and additional modification, e.g. multiple substitutions. In the absence of: (a) rational and predictable scheme for making any polypeptide comprising as little as two contiguous amino acids of SEQ ID NO:1, any polypeptide comprising as little as two contiguous amino acids of SEQ ID NO:1 and having a length of 18 contiguous amino acids, or having at least 70% sequence identity to SEQ ID NO:1, wherein the polypeptide has the function of directing protein decay and (b) a correlation between structure and the function of directing protein decay, the specification provides insufficient guidance as to which of the essentially infinite possible choices is likely to be successful. One of skill in the art would have to test these infinite possible polypeptides to determine which polypeptides have the function of directing protein decay. While enablement is not precluded by the necessity for routine screening, if a large amount of screening is required, as is the case herein, the specification must provide a reasonable amount of guidance which respect to the direction in which the experimentation should proceed so that a reasonable number of species can be selected for testing. In view of the fact that such guidance has not been provided in the instant specification, it would require undue experimentation to enable the full scope of the claims. The state of prior art, the relative skill of those in the art, and predictability or unpredictability of the art. Since the amino acid sequence of the protein determines its structural and functional properties, predictability of which changes can be tolerated in a protein's amino acid sequence and obtain the desired activity requires a knowledge of and guidance with regard to which amino acids in the protein's sequence, if any, are tolerant of modification and which are conserved (i.e. expectedly intolerant to modification), and detailed knowledge of the ways in which the proteins' structure relates to its function. In the instant case, neither the specification or the art provide a correlation between structure and activity such that one of skill in the art can envision the structure of any polypeptides having the function of directing protein decay or predict said function of a polypeptide from its primary structure. In addition, the art does not provide any teaching or guidance as to (1) which amino acids within the polynucleotide of SEQ ID NO:1 or any fragment thereof that can be modified and which ones are conserved such that one of skill in the art can make the recited polypeptides having the function of directing protein decay, (2) which segments of the polypeptide of SEQ ID NO:1 that are essential for polypeptides having the function of directing protein decay, and (3) the general tolerance of the polypeptide of SEQ ID NO:1 to structural modifications and the extent of such tolerance. The art clearly teaches that changes in a protein's amino acid sequence to obtain the desired activity without any guidance/knowledge as to which amino acids in a protein are required for that activity is highly unpredictable. At the time of the invention there was a high level of unpredictability associated with altering a polypeptide sequence with an expectation that the polypeptide will maintain the desired activity. For example, Studer (Residue mutations and their impact on protein structure and function: detecting beneficial and pathogenic changes. Biochem. J. (2013) 449, 581–594. – cited previously on form PTO-892) teach that (1) protein engineers are frequently surprised by the range of effects caused by single mutations that they hoped would change only one specific and simple property in enzymes, (2) the often surprising results obtained by experiments where single mutations are made reveal how little is known about the rules of protein stability, and (3) the difficulties in designing de novo stable proteins with specific functions. The polypeptide comprising the amino acid sequence of SEQ ID NO:1 was not known in the prior art. A search of SEQ ID NO:1 did not result in polypeptides having any significant similarity, as shown below. PNG media_image1.png 744 1194 media_image1.png Greyscale Fransceus (J Ind Microbiol Biotechnol. 2017 May;44(4-5):687-695. – form PTO-892) reviews protein engineering techniques, such as random mutagenesis and recombination, directed evolution and iterative or combinatory saturation “hotspots”. Fransceus states that “a recurring problem, however, is choosing which amino acid positions should be targeted. Answering this question is not an easy feat and requires substantial insight in the relationship between an enzyme’s sequence or structure and its properties.” Sanavia (Computational and Structural Biotechnology Journal, Volume 18, 2020, Pages 1968-1979. – form PTO-892) discloses challenges in the prediction of protein stability in the occurrence of multiple mutations. “Multiple-point mutations are common variations of the protein sequence that may be needed in protein engineering when a single-point mutation is not enough to yield the desired stability change. Dealing with multiple-site variations adds another level of complexity beyond the prediction of the effect of a single variant on protein stability, since it requires the learning of many types of combinatorial effects”. The amount of direction or guidance presented and the existence of working examples. The specification is limited to an expression cassette, vector, or host cell comprising a polynucleotide encoding a fusion protein comprising (a) the polypeptide comprising the amino acid sequence of SEQ ID NO:1 and (b) a protein of interest, wherein the polypeptide of (a) directs decay of the protein of interest of (b). However, the speciation fails to provide any information as to (1) specific substrates associated with polypeptides having at least 70% sequence identity to SEQ ID NO: 1 or fragments thereof or (2) structural elements required in a polypeptide having the function of directing protein decay. No correlation between structure and function of having the function of directing protein decay has been presented. There is no information or guidance as to which amino acid residues in the polypeptide of SEQ ID NO: 1 that can be modified and which ones are to be conserved to create a polypeptide having the function of directing protein decay. Thus, in view of the overly broad scope of the claims, the lack of guidance and working examples provided in the specification, the high level of unpredictability of the prior art in regard to structural changes and their effect on function and the lack of knowledge about a correlation between structure and function, an undue experimentation would be necessary one having ordinary skill in the art to make and use the claimed invention in a manner reasonably correlated with the scope of the claims. The scope of the claims must bear a reasonable correlation with the scope of enablement (In re Fisher, 166 USPQ 19 24 (CCPA 1970)). Without sufficient guidance, determination of polypeptides having the desired biological characteristics recited in the claims are unpredictable and the experimentation left to those skilled in the art is unnecessarily, and improperly, extensive and undue. See In re Wands 858 F.2d 731, 8 USPQ2nd 1400 (Fed. Cir, 1988). Applicant's arguments filed August 19, 2026 have been fully considered but they are not persuasive. Applicant has not submitted any rebuttal for the rejection. Applicant’s rebuttal at pages 12-16 is directed to the written description rejection. The claims are directed to an expression cassette, vector or host cell comprising a fusion protein comprising (a) a polypeptide having unknown structure, wherein the polypeptide has the function of directing protein decay and (b) a protein of interest. The claims are not commensurate with the enablement provided by the disclosure with regard to the extremely large number of polypeptides having the function of directing protein decay. The claims are not commensurate with the enablement provided by the disclosure with regard to the extremely large number of polypeptides having the function of directing protein decay. In the absence of: (a) rational and predictable scheme for making any polypeptide comprising as little as two contiguous amino acids of SEQ ID NO:1, any polypeptide comprising as little as two contiguous amino acids of SEQ ID NO:1 and having a length of 18 contiguous amino acids, or having at least 70% sequence identity to SEQ ID NO:1, wherein the polypeptide has the function of directing protein decay and (b) a correlation between structure and the function of directing protein decay, the specification provides insufficient guidance as to which of the essentially infinite possible choices is likely to be successful. One of skill in the art would have to test these infinite possible polypeptides to determine which polypeptides have the function of directing protein decay. While enablement is not precluded by the necessity for routine screening, if a large amount of screening is required, as is the case herein, the specification must provide a reasonable amount of guidance which respect to the direction in which the experimentation should proceed so that a reasonable number of species can be selected for testing. In view of the fact that such guidance has not been provided in the instant specification, it would require undue experimentation to enable the full scope of the claims. Given this lack of description of the representative species encompassed by the genus of the claims, the specification fails to sufficiently describe the claimed invention in such full, clear, concise, and exact terms that a skilled artisan would recognize that applicants were in possession of the inventions of claims 1-3 and 5-26. Therefore, the rejection has been maintained. Claim Rejections - 35 USC § 102 In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status. The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action: A person shall be entitled to a patent unless – (a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale or otherwise available to the public before the effective filing date of the claimed invention. Claim(s) 1-3, 5-8, 11-12, 15-16, 19, 21-22, and 26 is/are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Lin (US 10,550,379 - form PTO-1449 or WO 2017/004022 – form PTO-892. US 10,550,379 is used for specific passages of Lin). MPEP 2111.01 states that ''[d]uring examination, the claims must be interpreted as broadly as their terms reasonably allow.'' The limitation “an amino acid sequence” recited in claim 1 has been broadly interpreted as any amino acid sequence comprising as little as two contiguous amino acids of SEQ ID NO:1. The limitation “fragment thereof…wherein the fragment is at least 18 contiguous amino acids long” has been broadly interpreted as any amino acid sequence comprising any two contiguous amino acids of SEQ ID NO:1 and has a length of at least 18 contiguous amino acids. Regarding claim 1, Lin discloses an expression cassette (Column 18, line 64 through Column 19, line 12) encoding a fusion protein, comprising a) a nucleotide sequence encoding (i) a degron comprising the amino acid sequence shown in SEQ ID NO: 1, directs protein decay, and b) a nucleotide sequence encoding a protein of interest, wherein nucleotide sequence a) and b) are fused together in frame (Column 1, lines 49-64 and claims 1 and 21). The degron of SEQ ID NO:1 of Lin comprises at least two contiguous amino acids of SEQ ID NO:1 of the instant application (see the sequence alignment below). The degron of SEQ ID NO:1 of Lin is 42 amino acids long and therefore reads on a fragment of at least 18 contiguous amino acid long comprising as little as two contiguous amino acids of SEQ ID NO:1 of the instant application (see the sequence alignment below). Regarding claim 2, the degron of Lin is located at the N-terminus or at the C-terminus (Column 2, lines 28-63 and claim 2). Regarding claims 3 and 21, the expression cassette further comprises a nucleotide sequence encoding a cleavable linker comprising a fused to the 5'- and/ or 3'-end of the nucleotide sequence encoding the fusion protein (Column 2, lines 28-63 and claim 1). The nucleotide sequence comprises components, such as a protease, tags, targeting sequence, that are fused in frame with the degron, protein of interest, and the cleavable linker (Column 1 line 49 through Column 3, line 18). Regarding claim 5, the level of the fusion protein reduces when expressed in a cell in comparison to a cell which expresses the protein of interest (abstract). Regarding claim 6, the expression cassette of Lin comprises at least 3 nucleotides and encodes a heterologous polypeptide, wherein said heterologous polypeptide is a linker cleavable by a protease (abstract and claim 1). Regarding claim 7, the expression cassette of Lin has uses various promoters such as a the constitutive CMV promoter (Column 30, lines 55-65). Regarding claim 8, the protein of interest expression cassette of Lin encodes a polypeptide selected from a group consisting of enzymes, receptors, hormones, membrane proteins, and membrane associated proteins (claim 18). Regarding claim 11, Lin discloses a vector comprising the expression cassette (Column 3, lines 29-39). Regarding claim 12, Lin discloses a host cell comprising the expression cassette (claim 28). Regarding claim 15, Lin discloses a mammalian host cell (Column 3, lines 53-58 and claim 29) Regarding claims 16 and 26, the host cell of Lin not express one or more sterol acyltransferases of SEQ ID NO:15 and/or 16 of the instant application. Regarding claim 19, the host cell of Lin comprises a vector comprising the expression cassette (Column 3, line 33 through Column 4, line 4). Regarding claim 22, the expression cassette of Lin has regulatory sequences that turned on or off gene expression in response to a chemical stimulus, which reads on an inducible expression control sequence inducible by small molecules (Column 28, lines 41-49). Therefore, the reference of Lin anticipates claims 1-3, 5-8, 11-12, 15-16, 19, 21-22, and 26. Applicant's arguments filed August 19, 2026 have been fully considered but they are not persuasive. (A) Applicant argues that the degron sequence of Lin does not anticipate the polypeptide fragment as claimed in the present application since the specification explicitly and unambitiously defines inter alia the term “fragment” according the paragraph [0077] of the application. (B) Applicant argues that even though the degron of Lin has a total length of 42 amino acids, the total length is irrelevant for the determination of a “fragment” of the claimed sequence since the degron of Lin comprise merely 2 amino acids of SEQ ID NO:1 of the instant application. Applicant argues that there is no evidence that a mere 2-amino acid overlap can exert the specific biological activity required by paragraph [0077]. This is not found persuasive. Paragraph [0077] recites “In a preferred embodiment..”. Although the claims are interpreted in light of the specification, limitations from the specification are not read into the claims. See In re Van Geuns, 988 F.2d 1181, 26 USPQ2d 1057 (Fed. Cir. 1993). “Though understanding the claim language may be aided by explanations contained in the written description, it is important not to import into a claim limitations that are not part of the claim. For example, a particular embodiment appearing in the written description may not be read into a claim when the claim language is broader than the embodiment….(“We have cautioned against reading limitations into a claim from the preferred embodiment described in the specification, even if it is the only embodiment described, absent clear disclaimer in the specification.”).”, see MPEP 2111.I. and MPEP 2111.II. In the instant case, claim 1 recites that the nucleotide sequence encodes (1) an amino acid sequence shown in SEQ ID NO:1 or (2) a fragment thereof… wherein the fragment is at least 18 contiguous amino acids long. The limitation “an amino acid sequence” recited in claim 1 has been broadly interpreted as any amino acid sequence comprising as little as two contiguous amino acids of SEQ ID NO:1. The limitation “fragment thereof…wherein the fragment is at least 18 contiguous amino acids long” has been broadly interpreted as any amino acid sequence comprising any two contiguous amino acids of SEQ ID NO:1 and has a length of at least 18 contiguous amino acids. (A) The degron of Lin comprises at least two contiguous amino acids of SEQ ID NO:1 of the instant application (see the sequence alignment below), satisfying the structural requirement of the encoded amino acid sequence of claim 1 a). (2) The degron of Lin comprises at least two contiguous amino acids of SEQ ID NO:1 of the instant application and has a length of at least 18 contiguous amino acids, satisfying the structural requirement of the encoded amino acid sequence of claim 1 a). Claim 1 does not recite that the fragment comprises of 18 contiguous amino acids of SEQ ID NO:1. Therefore, the degron does not need to be a fragment comprising 18 contiguous amino acids of SEQ ID NO:1. (B) The degron of Lin directs protein decay, satisfying the functional requirement of the encoded amino acid sequence of claim 1 a). Claim 1 does not recite that the encoded amino acid of claim 1 a) has identical function as the polypeptide having the amino acid sequence of SEQ ID NO:1 of the instant application. Therefore, the claims are anticipated by Lin. Hence the rejection has been maintained. Claim Rejections - 35 USC § 103 The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action: A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made. The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows: 1. Determining the scope and contents of the prior art. 2. Ascertaining the differences between the prior art and the claims at issue. 3. Resolving the level of ordinary skill in the pertinent art. 4. Considering objective evidence present in the application indicating obviousness or nonobviousness. This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention. Claim(s) 1-3, 5-9, 11-23, 25, and 26 is/are rejected under 35 U.S.C. 103 as being unpatentable over Lin (US 10,550,379 - form PTO-1449 or WO 2017/004022 – form PTO-892. US 10,550,379 is used for specific passages of Lin) and Schulze (WO 2019/197327 – form PTO-892). MPEP 2111.01 states that ''[d]uring examination, the claims must be interpreted as broadly as their terms reasonably allow.'' The limitation “an amino acid sequence” recited in claim 1 has been broadly interpreted as any amino acid sequence comprising as little as two contiguous amino acids of SEQ ID NO:1. The limitation “fragment thereof…wherein the fragment is at least 18 contiguous amino acids long” has been broadly interpreted as any amino acid sequence comprising any two contiguous amino acids of SEQ ID NO:1 and has a length of at least 18 contiguous amino acids. Regarding claim 1, Lin discloses an expression cassette (Column 18, line 64 through Column 19, line 12) encoding a fusion protein, comprising a) a nucleotide sequence encoding (i) a degron comprising the amino acid sequence shown in SEQ ID NO: 1, directs protein decay, and b) a nucleotide sequence encoding a protein of interest, wherein nucleotide sequence a) and b) are fused together in frame (Column 1, lines 49-64 and claims 1 and 21). The degron of SEQ ID NO:1 of Lin comprises at least two contiguous amino acids of SEQ ID NO:1 of the instant application (see the sequence alignment below). The degron of SEQ ID NO:1 of Lin is 42 amino acids long and therefore reads on a fragment of at least 18 amino acid long comprising as little as two contiguous amino acids of SEQ ID NO:1 of the instant application (see the sequence alignment below). Regarding claim 2, the degron of Lin is located at the N-terminus or at the C-terminus (Column 2, lines 28-63 and claim 2). Regarding claims 3 and 21, the expression cassette further comprises a nucleotide sequence encoding a cleavable linker fused to the 5'- and/ or 3'-end of the nucleotide sequence encoding the fusion protein (Column 2, lines 28-63 and claim 1). The nucleotide sequence comprises components, such as a protease, tags, targeting sequence, that are fused in frame with the degron, protein of interest, and the cleavable linker (Column 1 line 49 through Column 3, line 18). Regarding claim 5, the level of the fusion protein reduces when expressed in a cell in comparison to a cell which expresses the protein of interest (abstract). Regarding claim 6, the expression cassette of Lin comprises at least 3 nucleotides and encodes a heterologous polypeptide, wherein said heterologous polypeptide is a linker cleavable by a protease (abstract and claim 1). Regarding claim 7, the expression cassette of Lin has uses various promoters such as a the constitutive CMV promoter (Column 30, lines 55-65). Regarding claim 8, the protein of interest expression cassette of Lin encodes a polypeptide selected from a group consisting of enzymes, receptors, hormones, membrane proteins, and membrane associated proteins (claim 18). Regarding claim 11, Lin discloses a vector comprising the expression cassette (Column 3, lines 29-39). Regarding claim 12, Lin discloses a host cell comprising the expression cassette (claim 28). Regarding claim 15, Lin discloses a mammalian host cell (Column 3, lines 53-58 and claim 29) Regarding claims 16 and 26, the host cell of Lin not express one or more sterol acyltransferases of SEQ ID NO:15 and/or 16 of the instant application. Regarding claim 19, the host cell of Lin comprises a vector comprising the expression cassette (Column 3, line 33 through Column 4, line 4). Regarding claim 22, the expression cassette of Lin has regulatory sequences that turned on or off gene expression in response to a chemical stimulus, which reads on an inducible expression control sequence inducible by small molecules (Column 28, lines 41-49). However, Lin does not disclose a fusion protein wherein the protein of interest is a lanosterol synthase (ERG7) having the amino acid sequence of SEQ ID NO:3 and encoded by SEQ ID NO:4 nor a method of producing a triterpenoid. Regarding claims 9, 13-14, 23, and 25, Schulze discloses an engineered host cell to repress the lanosterol synthase (ERG7) having the amino acid sequence of SEQ ID NO:9 and a polynucleotide encoding said lanosterol synthase ([0015] and claim 4). The lanosterol synthase of SEQ ID NO:9 of Schulze is identical to the lanosterol synthase of SEQ ID NO:3 of the instant application and the polynucleotide encoding the lanosterol synthase of SEQ ID NO:9 of Schulze is identical to the polynucleotide of SEQ ID NO:4 of the instant application (see the sequence alignments below). Regarding claims 15 and 20, Schulze discloses a yeast host engineered to repress the lanosterol synthase (claims 1 and 10). Regarding claims 16 and 26, the yeast host cell of Schulze does not express one or more sterol acyltransferases of SEQ ID NO:15 and/or 16 of the instant application. Regarding claim 17, Schulze discloses using a cytochrome P450 reductase and a cytochrome P450 monooxygenase to produce triterpenes in yeast ([006]). Regarding claim 18, Schulze discloses a method of increasing the yield of a triterpenoid in a host cell or yeast host cell, wherein the lanosterol synthase (ERG7) is repressed (claims 1 and 4). Therefore, in combining the teachings of Lin and Schulze, it would have been obvious to one having ordinary skill in the art before the time the claimed invention was effectively filed to make an expression cassette, vector, or yeast host cell comprising a polynucleotide encoding a fusion protein comprising (a) the degron of Lin and (b) lanosterol synthase (ERG7), and a cleavable linker, and a method of producing a triterpenoid using said yeast host cell. One of ordinary skill in the art at the time the invention was made would have been motivated to do so in order to control the enzymatic activity of lanosterol synthase to increase the yield of triterpenoid production. One of ordinary skill in the art would have had a reasonable expectation of success since Lin discloses a polynucleotide encoding a fusion protein comprising a degron and a protein of interest, wherein the degron decays the protein of interest and Schulze discloses repressing lanosterol synthase (ERG7) in order to increase the yield of triterpenoid production. Therefore, the above references render claims 1-3, 5-9, 11-23, 25, and 26 prima facie obvious. Applicant's arguments filed August 19, 2026 have been fully considered but they are not persuasive. Applicant argues that the claims are not obvious because the degron sequence of Lin is different than the amino acid sequence of SEQ ID NO:1 of the instant application or a fragment thereof. This is not found persuasive. The claims do not recite a limitation that the amino acid sequence that direct protein decay have identical structure to the amino acid sequence of SEQ ID NO:1 of the instant application. As discussed above, the degron of Lin comprises at least two contiguous amino acids of SEQ ID NO:1 of the instant application (see the sequence alignment below), satisfying the structural requirement of the encoded amino acid sequence of claim 1 a). The degron of Lin also comprises at least two contiguous amino acids of SEQ ID NO:1 of the instant application and has a length of at least 18 contiguous amino acids, satisfying the structural requirement of the encoded amino acid sequence of claim 1 a). Applicant argues that the claims are not obvious because the mechanism of Lin is suppression of the expression of the protein of interest in the presence of a protease inhibitor and Schulze relates to repress ERG7 by the insertion of the CTR3-promoter and/or the addition of copper sulfate CuSO4 and therefore combining the teachings of Yin and Schulze would change the fundamental principle operation of both references. This is not found persuasive. Lin discloses a fusion protein construct to control the production of a protein of interest such that degradation of a protein of interest is controllable. Schulze is relied on its teaching of repressing expression of ERG7, which is involved in a competitive isoprenoid pathway ([004]-[005]). One of ordinary skill in the art at the time the invention was made would have been motivated use ERG7 as the protein of interest in order to control the enzymatic activity of lanosterol synthase to increase the yield of triterpenoid production. The modified cassette of Lin comprising the fusion protein comprising ERG7 as the protein of interest allows for controlling the activity of ERG7. The modified cassette of Lin comprising the fusion protein comprising ERG7 not comprise or utilize CTR3-promoter and/or the addition of copper sulfate CuSO4. Therefore, the modified expression cassette of Lin does not have a change in fundamental principle of operation. Hence the rejection is maintained. Conclusion Claims 1-26 are pending. Claims 1-3 and 5-26 are rejected. Claim 4 is objected to as being dependent upon a rejected base claim, but would be allowable if rewritten in independent form including all of the limitations of the base claim and any intervening claims. Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a). A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action. Any inquiry concerning this communication or earlier communications from the examiner should be directed to YONG D PAK whose telephone number is (571)272-0935. The examiner can normally be reached M-Th: 5:30 am - 3:30 pm. Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice. If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Robert Mondesi can be reached on 408-918-7584. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300. Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000. /YONG D PAK/Primary Examiner, Art Unit 1652 Sequence alignment between the peptide of SEQ ID NO:1 of the instant application (“Qy”) and the degron of SEQ ID NO:1 of Lin (“Db”) Title: US-18-709-148A-1 Perfect score: 155 Sequence: 1 LRLEQVLVLVLEQFCLKVKNYSLVLSQFWLN 31 Scoring table: BLOSUM62 Gapop 10.0 , Gapext 0.5 Searched: 1 seqs, 42 residues Total number of hits satisfying chosen parameters: 1 Minimum DB seq length: 0 Maximum DB seq length: inf Post-processing: Minimum Match 0% Maximum Match 100% Listing first 1 summaries Database : AASEQ2_03172026_154334.pep:* SUMMARIES % Result Query No. Score Match Length DB ID Description ---------------------------------------------------------------------------- 1 19 12.3 42 1 AASEQ2_03172026_154334 ALIGNMENTS RESULT 1 AASEQ2_03172026_154334 Query Match 12.3%; Score 19; DB 1; Length 42; Best Local Similarity 30.0%; Matches 3; Conservative 3; Mismatches 4; Indels 0; Gaps 0; Qy 7 LVLVLEQFCL 16 :: | :|| Db 31 VLAALAAYCL 40 Sequence alignment between the lanosterol synthase (ERG7) of SEQ ID NO:3 of the instant application (“Qy”) and the lanosterol synthase (ERG7) of SEQ ID NO:9 of Schulze (“Db”) BGW52013 ID BGW52013 standard; DNA; 2196 BP. XX AC BGW52013; XX DT 12-DEC-2019 (first entry) XX DE Saccharomyces cerevisiae lanosterol synthase (ERG7) DNA, SEQ ID 20. XX KW ERG7; ds; expression; gene; genetic engineering; lanosterol synthase. XX OS Saccharomyces cerevisiae. XX FH Key Location/Qualifiers FT CDS 1..2196 FT /*tag= a FT /product= " lanosterol synthase (ERG7) protein" XX CC PN WO2019197327-A1. XX CC PD 17-OCT-2019. XX CC PF 08-APR-2019; 2019WO-EP058789. XX PR 09-APR-2018; 2018EP-00166374. XX CC PA (FRAU ) FRAUNHOFER GES FOERDERUNG ANGEWANDTEN EV. CC PA (UYMU ) UNIV MUENSTER WESTFAELISCHE WILHELMS. XX CC PI Schulze Gronover C, Boje Mueller LG, Van Deenen N, Broeker JN; XX DR WPI; 2019-86338C/83. DR P-PSDB; BGW52002. XX CC PT Increasing yield of one of oxidosqualene, triterpenes and/or CC PT triterpenoids in host cell involves engineering host cell to overexpress CC PT a 3-hydroxy-3-methylglutaryl-coenzyme A reductase comprising an amino CC PT acids. XX CC PS Example 2; SEQ ID NO 20; 96pp; English. XX CC The invention relates to a novel method of increasing the yield of at CC least one of oxidosqualene, triterpenes and triterpenoids in a host cell. CC The method involves engineering host cell to overexpress a 3-hydroxy-3- CC methylglutaryl-coenzyme A reductase comprising an amino acid shown in SEQ CC ID 1 (see BGW51994). The invention also provides a recombinant host cell CC for manufacturing one of oxidosqualene, triterpenes and triterpenoids. CC The method is useful for manufacturing one of oxidosqualene, triterpenes CC and triterpenoids. XX SQ Sequence 2196 BP; 652 A; 448 C; 476 G; 620 T; 0 U; 0 Other; Query Match 100.0%; Score 3939; Length 731; Best Local Similarity 100.0%; Matches 731; Conservative 0; Mismatches 0; Indels 0; Gaps 0; Qy 1 MTEFYSDTIGLPKTDPRLWRLRTDELGRESWEYLTPQQAANDPPSTFTQWLLQDPKFPQP 60 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 1 MTEFYSDTIGLPKTDPRLWRLRTDELGRESWEYLTPQQAANDPPSTFTQWLLQDPKFPQP 60 Qy 61 HPERNKHSPDFSAFDACHNGASFFKLLQEPDSGIFPCQYKGPMFMTIGYVAVNYIAGIEI 120 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 61 HPERNKHSPDFSAFDACHNGASFFKLLQEPDSGIFPCQYKGPMFMTIGYVAVNYIAGIEI 120 Qy 121 PEHERIELIRYIVNTAHPVDGGWGLHSVDKSTVFGTVLNYVILRLLGLPKDHPVCAKARS 180 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 121 PEHERIELIRYIVNTAHPVDGGWGLHSVDKSTVFGTVLNYVILRLLGLPKDHPVCAKARS 180 Qy 181 TLLRLGGAIGSPHWGKIWLSALNLYKWEGVNPAPPETWLLPYSLPMHPGRWWVHTRGVYI 240 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 181 TLLRLGGAIGSPHWGKIWLSALNLYKWEGVNPAPPETWLLPYSLPMHPGRWWVHTRGVYI 240 Qy 241 PVSYLSLVKFSCPMTPLLEELRNEIYTKPFDKINFSKNRNTVCGVDLYYPHSTTLNIANS 300 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 241 PVSYLSLVKFSCPMTPLLEELRNEIYTKPFDKINFSKNRNTVCGVDLYYPHSTTLNIANS 300 Qy 301 LVVFYEKYLRNRFIYSLSKKKVYDLIKTELQNTDSLCIAPVNQAFCALVTLIEEGVDSEA 360 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 301 LVVFYEKYLRNRFIYSLSKKKVYDLIKTELQNTDSLCIAPVNQAFCALVTLIEEGVDSEA 360 Qy 361 FQRLQYRFKDALFHGPQGMTIMGTNGVQTWDCAFAIQYFFVAGLAERPEFYNTIVSAYKF 420 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 361 FQRLQYRFKDALFHGPQGMTIMGTNGVQTWDCAFAIQYFFVAGLAERPEFYNTIVSAYKF 420 Qy 421 LCHAQFDTECVPGSYRDKRKGAWGFSTKTQGYTVADCTAEAIKAIIMVKNSPVFSEVHHM 480 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 421 LCHAQFDTECVPGSYRDKRKGAWGFSTKTQGYTVADCTAEAIKAIIMVKNSPVFSEVHHM 480 Qy 481 ISSERLFEGIDVLLNLQNIGSFEYGSFATYEKIKAPLAMETLNPAEVFGNIMVEYPYVEC 540 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 481 ISSERLFEGIDVLLNLQNIGSFEYGSFATYEKIKAPLAMETLNPAEVFGNIMVEYPYVEC 540 Qy 541 TDSSVLGLTYFHKYFDYRKEEIRTRIRIAIEFIKKSQLPDGSWYGSWGICFTYAGMFALE 600 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 541 TDSSVLGLTYFHKYFDYRKEEIRTRIRIAIEFIKKSQLPDGSWYGSWGICFTYAGMFALE 600 Qy 601 ALHTVGETYENSSTVRKGCDFLVSKQMKDGGWGESMKSSELHSYVDSEKSLVVQTAWALI 660 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 601 ALHTVGETYENSSTVRKGCDFLVSKQMKDGGWGESMKSSELHSYVDSEKSLVVQTAWALI 660 Qy 661 ALLFAEYPNKEVIDRGIDLLKNRQEESGEWKFESVEGVFNHSCAIEYPSYRFLFPIKALG 720 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 661 ALLFAEYPNKEVIDRGIDLLKNRQEESGEWKFESVEGVFNHSCAIEYPSYRFLFPIKALG 720 Qy 721 MYSRAYETHTL 731 ||||||||||| Db 721 MYSRAYETHTL 731 Sequence alignment between the polynucleotide SEQ ID NO:4 of the instant application (“Qy”) and the polynucleotide encoding the lanosterol synthase (ERG7) of SEQ ID NO:9 of Schulze (“Db”) BGW52013 ID BGW52013 standard; DNA; 2196 BP. XX AC BGW52013; XX DT 12-DEC-2019 (first entry) XX DE Saccharomyces cerevisiae lanosterol synthase (ERG7) DNA, SEQ ID 20. XX KW ERG7; ds; expression; gene; genetic engineering; lanosterol synthase. XX OS Saccharomyces cerevisiae. XX FH Key Location/Qualifiers FT CDS 1..2196 FT /*tag= a FT /product= " lanosterol synthase (ERG7) protein" XX CC PN WO2019197327-A1. XX CC PD 17-OCT-2019. XX CC PF 08-APR-2019; 2019WO-EP058789. XX PR 09-APR-2018; 2018EP-00166374. XX CC PA (FRAU ) FRAUNHOFER GES FOERDERUNG ANGEWANDTEN EV. CC PA (UYMU ) UNIV MUENSTER WESTFAELISCHE WILHELMS. XX CC PI Schulze Gronover C, Boje Mueller LG, Van Deenen N, Broeker JN; XX DR WPI; 2019-86338C/83. DR P-PSDB; BGW52002. XX CC PT Increasing yield of one of oxidosqualene, triterpenes and/or CC PT triterpenoids in host cell involves engineering host cell to overexpress CC PT a 3-hydroxy-3-methylglutaryl-coenzyme A reductase comprising an amino CC PT acids. XX CC PS Example 2; SEQ ID NO 20; 96pp; English. XX CC The invention relates to a novel method of increasing the yield of at CC least one of oxidosqualene, triterpenes and triterpenoids in a host cell. CC The method involves engineering host cell to overexpress a 3-hydroxy-3- CC methylglutaryl-coenzyme A reductase comprising an amino acid shown in SEQ CC ID 1 (see BGW51994). The invention also provides a recombinant host cell CC for manufacturing one of oxidosqualene, triterpenes and triterpenoids. CC The method is useful for manufacturing one of oxidosqualene, triterpenes CC and triterpenoids. XX SQ Sequence 2196 BP; 652 A; 448 C; 476 G; 620 T; 0 U; 0 Other; Query Match 100.0%; Score 2196; Length 2196; Best Local Similarity 100.0%; Matches 2196; Conservative 0; Mismatches 0; Indels 0; Gaps 0; Qy 1 ATGACAGAATTTTATTCTGACACAATCGGTCTACCAAAGACAGATCCACGTCTTTGGAGA 60 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 1 ATGACAGAATTTTATTCTGACACAATCGGTCTACCAAAGACAGATCCACGTCTTTGGAGA 60 Qy 61 CTGAGAACTGATGAGCTAGGCCGAGAAAGCTGGGAATATTTAACCCCTCAGCAAGCCGCA 120 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 61 CTGAGAACTGATGAGCTAGGCCGAGAAAGCTGGGAATATTTAACCCCTCAGCAAGCCGCA 120 Qy 121 AACGACCCACCATCCACTTTCACGCAGTGGCTTCTTCAAGATCCCAAATTTCCTCAACCT 180 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 121 AACGACCCACCATCCACTTTCACGCAGTGGCTTCTTCAAGATCCCAAATTTCCTCAACCT 180 Qy 181 CATCCAGAAAGAAATAAGCATTCACCAGATTTTTCAGCCTTCGATGCGTGTCATAATGGT 240 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 181 CATCCAGAAAGAAATAAGCATTCACCAGATTTTTCAGCCTTCGATGCGTGTCATAATGGT 240 Qy 241 GCATCTTTTTTCAAACTGCTTCAAGAGCCTGACTCAGGTATTTTTCCGTGTCAATATAAA 300 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 241 GCATCTTTTTTCAAACTGCTTCAAGAGCCTGACTCAGGTATTTTTCCGTGTCAATATAAA 300 Qy 301 GGACCCATGTTCATGACAATCGGTTACGTAGCCGTAAACTATATCGCCGGTATTGAAATT 360 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 301 GGACCCATGTTCATGACAATCGGTTACGTAGCCGTAAACTATATCGCCGGTATTGAAATT 360 Qy 361 CCTGAGCATGAGAGAATAGAATTAATTAGATACATCGTCAATACAGCACATCCGGTTGAT 420 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 361 CCTGAGCATGAGAGAATAGAATTAATTAGATACATCGTCAATACAGCACATCCGGTTGAT 420 Qy 421 GGTGGCTGGGGTCTACATTCTGTTGACAAATCCACCGTGTTTGGTACAGTATTGAACTAT 480 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 421 GGTGGCTGGGGTCTACATTCTGTTGACAAATCCACCGTGTTTGGTACAGTATTGAACTAT 480 Qy 481 GTAATCTTACGTTTATTGGGTCTACCCAAGGACCACCCGGTTTGCGCCAAGGCAAGAAGC 540 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 481 GTAATCTTACGTTTATTGGGTCTACCCAAGGACCACCCGGTTTGCGCCAAGGCAAGAAGC 540 Qy 541 ACATTGTTAAGGTTAGGCGGTGCTATTGGATCCCCTCACTGGGGAAAAATTTGGCTAAGT 600 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 541 ACATTGTTAAGGTTAGGCGGTGCTATTGGATCCCCTCACTGGGGAAAAATTTGGCTAAGT 600 Qy 601 GCACTAAACTTGTATAAATGGGAAGGTGTGAACCCTGCCCCTCCTGAAACTTGGTTACTT 660 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 601 GCACTAAACTTGTATAAATGGGAAGGTGTGAACCCTGCCCCTCCTGAAACTTGGTTACTT 660 Qy 661 CCATATTCACTGCCCATGCATCCGGGGAGATGGTGGGTTCATACTAGAGGTGTTTACATT 720 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 661 CCATATTCACTGCCCATGCATCCGGGGAGATGGTGGGTTCATACTAGAGGTGTTTACATT 720 Qy 721 CCGGTCAGTTACCTGTCATTGGTCAAATTTTCTTGCCCAATGACTCCTCTTCTTGAAGAA 780 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 721 CCGGTCAGTTACCTGTCATTGGTCAAATTTTCTTGCCCAATGACTCCTCTTCTTGAAGAA 780 Qy 781 CTGAGGAATGAAATTTACACTAAACCGTTTGACAAGATTAACTTCTCCAAGAACAGGAAT 840 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 781 CTGAGGAATGAAATTTACACTAAACCGTTTGACAAGATTAACTTCTCCAAGAACAGGAAT 840 Qy 841 ACCGTATGTGGAGTAGACCTATATTACCCCCATTCTACTACTTTGAATATTGCGAACAGC 900 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 841 ACCGTATGTGGAGTAGACCTATATTACCCCCATTCTACTACTTTGAATATTGCGAACAGC 900 Qy 901 CTTGTAGTATTTTACGAAAAATACCTAAGAAACCGGTTCATTTACTCTCTATCCAAGAAG 960 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 901 CTTGTAGTATTTTACGAAAAATACCTAAGAAACCGGTTCATTTACTCTCTATCCAAGAAG 960 Qy 961 AAGGTTTATGATCTAATCAAAACGGAGTTACAGAATACTGATTCCTTGTGTATAGCACCT 1020 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 961 AAGGTTTATGATCTAATCAAAACGGAGTTACAGAATACTGATTCCTTGTGTATAGCACCT 1020 Qy 1021 GTTAACCAGGCGTTTTGCGCACTTGTCACTCTTATTGAAGAAGGGGTAGACTCGGAAGCG 1080 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 1021 GTTAACCAGGCGTTTTGCGCACTTGTCACTCTTATTGAAGAAGGGGTAGACTCGGAAGCG 1080 Qy 1081 TTCCAGCGTCTCCAATATAGGTTCAAGGATGCATTGTTCCATGGTCCACAGGGTATGACC 1140 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 1081 TTCCAGCGTCTCCAATATAGGTTCAAGGATGCATTGTTCCATGGTCCACAGGGTATGACC 1140 Qy 1141 ATTATGGGAACAAATGGTGTGCAAACCTGGGATTGTGCGTTTGCCATTCAATACTTTTTC 1200 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 1141 ATTATGGGAACAAATGGTGTGCAAACCTGGGATTGTGCGTTTGCCATTCAATACTTTTTC 1200 Qy 1201 GTCGCAGGCCTCGCAGAAAGACCTGAATTCTATAACACAATTGTCTCTGCCTATAAATTC 1260 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 1201 GTCGCAGGCCTCGCAGAAAGACCTGAATTCTATAACACAATTGTCTCTGCCTATAAATTC 1260 Qy 1261 TTGTGTCATGCTCAATTTGACACCGAGTGCGTTCCAGGTAGTTATAGGGATAAGAGAAAG 1320 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 1261 TTGTGTCATGCTCAATTTGACACCGAGTGCGTTCCAGGTAGTTATAGGGATAAGAGAAAG 1320 Qy 1321 GGGGCTTGGGGCTTCTCAACAAAAACACAGGGCTATACAGTGGCAGATTGCACTGCAGAA 1380 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 1321 GGGGCTTGGGGCTTCTCAACAAAAACACAGGGCTATACAGTGGCAGATTGCACTGCAGAA 1380 Qy 1381 GCAATTAAAGCCATCATCATGGTGAAAAACTCTCCCGTCTTTAGTGAAGTACACCATATG 1440 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 1381 GCAATTAAAGCCATCATCATGGTGAAAAACTCTCCCGTCTTTAGTGAAGTACACCATATG 1440 Qy 1441 ATTAGCAGTGAACGTTTATTTGAAGGCATTGATGTGTTATTGAACCTACAAAACATCGGA 1500 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 1441 ATTAGCAGTGAACGTTTATTTGAAGGCATTGATGTGTTATTGAACCTACAAAACATCGGA 1500 Qy 1501 TCTTTTGAATATGGTTCCTTTGCAACCTATGAAAAAATCAAGGCCCCACTAGCAATGGAA 1560 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 1501 TCTTTTGAATATGGTTCCTTTGCAACCTATGAAAAAATCAAGGCCCCACTAGCAATGGAA 1560 Qy 1561 ACCTTGAATCCTGCTGAAGTTTTTGGTAACATAATGGTAGAATACCCATACGTGGAATGT 1620 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 1561 ACCTTGAATCCTGCTGAAGTTTTTGGTAACATAATGGTAGAATACCCATACGTGGAATGT 1620 Qy 1621 ACTGATTCATCCGTTCTGGGGTTGACATATTTTCACAAGTACTTCGACTATAGGAAAGAG 1680 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 1621 ACTGATTCATCCGTTCTGGGGTTGACATATTTTCACAAGTACTTCGACTATAGGAAAGAG 1680 Qy 1681 GAAATACGTACACGCATCAGAATCGCCATCGAATTCATAAAAAAATCTCAATTACCAGAT 1740 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 1681 GAAATACGTACACGCATCAGAATCGCCATCGAATTCATAAAAAAATCTCAATTACCAGAT 1740 Qy 1741 GGAAGTTGGTATGGAAGCTGGGGTATTTGTTTTACATATGCCGGTATGTTTGCATTGGAG 1800 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 1741 GGAAGTTGGTATGGAAGCTGGGGTATTTGTTTTACATATGCCGGTATGTTTGCATTGGAG 1800 Qy 1801 GCATTACACACCGTGGGGGAGACCTATGAGAATTCCTCAACGGTAAGAAAAGGTTGCGAC 1860 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 1801 GCATTACACACCGTGGGGGAGACCTATGAGAATTCCTCAACGGTAAGAAAAGGTTGCGAC 1860 Qy 1861 TTCTTGGTCAGTAAACAGATGAAGGATGGCGGTTGGGGGGAATCAATGAAGTCCAGTGAA 1920 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 1861 TTCTTGGTCAGTAAACAGATGAAGGATGGCGGTTGGGGGGAATCAATGAAGTCCAGTGAA 1920 Qy 1921 TTACATAGTTATGTGGATAGTGAAAAATCGCTAGTCGTTCAAACCGCATGGGCGCTAATT 1980 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 1921 TTACATAGTTATGTGGATAGTGAAAAATCGCTAGTCGTTCAAACCGCATGGGCGCTAATT 1980 Qy 1981 GCACTTCTTTTCGCTGAATATCCTAATAAAGAAGTCATCGACCGCGGTATTGACCTTTTA 2040 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 1981 GCACTTCTTTTCGCTGAATATCCTAATAAAGAAGTCATCGACCGCGGTATTGACCTTTTA 2040 Qy 2041 AAAAATAGACAAGAAGAATCCGGGGAATGGAAATTTGAAAGTGTAGAAGGTGTTTTCAAC 2100 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 2041 AAAAATAGACAAGAAGAATCCGGGGAATGGAAATTTGAAAGTGTAGAAGGTGTTTTCAAC 2100 Qy 2101 CACTCTTGTGCAATTGAATACCCAAGTTATCGATTCTTATTCCCTATTAAGGCATTAGGT 2160 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Db 2101 CACTCTTGTGCAATTGAATACCCAAGTTATCGATTCTTATTCCCTATTAAGGCATTAGGT 2160 Qy 2161 ATGTACAGCAGGGCATATGAAACACATACGCTTTAA 2196 |||||||||||||||||||||||||||||||||||| Db 2161 ATGTACAGCAGGGCATATGAAACACATACGCTTTAA 2196