Engineering the synthetic β-alanine pathway in Komagataella phaffii for conversion of methanol into 3-hydroxypropionic acid

Table 4 Variable sgRNA targets custom-designed in this study

Target name	Locus	Sequence (5′→3′)	Expression cassette
pTEF1-α^UP_sgRNA1	PP7435_Chr1-1535	AACAACACTAAACTACCTTG	P_FDH1_BAPAT_Bc_ScCYC1tt
pTEF1-α^UP_sgRNA2	PP7435_Chr1-1535	TTAAGGATGTGTAGTGTCAA	P_FDH1_BAPAT_Bc_ScCYC1tt
pFLD1^UP_sgRNA1	PP7435_Chr3-0140	TGCTAATGGTAGTTATCCAA	P_POR1-YDFG_Ec-TDH3tt P_PDC1-YDFG_Ec-TDH3tt
pFLD1^UP_sgRNA2	PP7435_Chr3-0140	CTATAGGATAAAAACAGGAG	P_POR1-YDFG_Ec-TDH3tt P_PDC1-YDFG_Ec-TDH3tt
RGI2_sgRNA1	PP7435_Chr1-0725	TCTCAACGTATTTATATGGT	P_AOX1-PAND_Tc-RPS3tt
RGI2_sgRNA2	PP7435_Chr1-0725	ATGAAGCCACTTCAACTACG	P_AOX1-PAND_Tc-RPS3tt
pGAP^UP_sgRNA1	PP7435_Chr2-0858	ATCGATAATAGTCGCATGTG	P_AOX1-PAND_Tc-RPS3tt
pGAP^UP_sgRNA2	PP7435_Chr2-0858	CGTTAGGTCAGTGATGACAA	P_AOX1-PAND_Tc-RPS3tt
pAOX1^UP_sgRNA1	PP7435_Chr4-0130	ATTGTGAAATAGACGCAGAT	P_FDH1-FDH(V9)_Pse-TDH3tt
pAOX1^UP_sgRNA2	PP7435_Chr4-0130	GCAGTCGATCTCAAAAGCAA	P_FDH1-FDH(V9)_Pse-TDH3tt

ISSN: 1475-2859