Parts
Part ID Name / Vector Description / Function
BBa_25A067ED p414-TEF1p-Cas9-CYC1t It carries TEF1p promoter and CYC1 terminator, providing the core framework for Cas9 expression in CRISPR‑Cas9 systems. As the basic backbone for fungal CRISPR‑Cas9 vectors, it accepts inserts like NAT, AMA1 and sgRNA cassettes after linearization. It supports targeted gene editing in fungi (e.g., Aureobasidium melanogenum), laying a foundation for gene function verification and metabolic engineering of non‑model fungi.
BBa_25TR75TF Autonomous Maintenance in Aspergillus 1 (AMA1) AMA1 is a DNA sequence isolated from Aspergillus nidulans, which enables plasmids to replicate autonomously in filamentous fungi (such as Aspergillus flavus). It functions as a core element of the plasmid "backbone" responsible for origin of replication activity.
BBa_256AEFB2 phosphoribosylaminoimidazole carboxylase ADE2 sgRNA in Aureobasidium melanogenum The function of the Ade2 gRNA in the CRISPR‑Cas9‑Am system is to recognize and bind the specific target sequence of the Ade2 gene in Aureobasidium melanogenum, then guide the Cas9 nuclease to accurately locate and cleave the Ade2 locus, ultimately mediating knockout of the gene. This enables verification of gene‑editing feasibility of the CRISPR‑Cas9‑Am vector through a colony color phenotype (wild‑type white → knockout red).
BBa_252UGU5Z CRISPR-Cas9-Am The CRISPR‑Cas9‑Am knockout vector is a fungal‑specific gene editing tool designed for Aureobasidium melanogenum to overcome weak genetic background research and the lack of effective editing systems. It enables precise gene editing via Cas9 guided by sgRNA, supporting functional gene studies and metabolic engineering. The vector carries the NAT resistance gene for positive screening and the AMA1 autonomous replication sequence for stable plasmid maintenance. Cas9 expression is driven by the strong TEF1 promoter and terminated by the CYC1 terminator, ensuring efficient protein production. sgRNA expression is controlled by the U6 promoter/terminator, enabling precise target recognition. A replaceable sgRNA region allows flexible editing of different genes without reconstructing the vector, making it versatile and efficient for fungal gene editing.
BBa_25MXN531 f14a-nat-loxp Template for PCR amplification of Nourseothricin (NAT) resistance gene to provide a selection marker for the vector.
BBa_25FPRM5L pNat-LoxP-rDNA-P.p Das Heterologously expresses Pichia pastoris Das gene to enhance methanol metabolism flux in Aureobasidium pullulans.
BBa_25QPWVAX pNAT-LoxP-rDNA Basic plasmid for subcellular localization; contains NAT resistance gene, LoxP site and rDNA sequence for inserting fluorescent labeling cassettes.
BBa_25TNP0I3 pHPT-LoxP-rDNA Basic plasmid for subcellular localization vector; contains HPT resistance gene, LoxP site and rDNA sequence for inserting fluorescent labeling elements.
BBa_25DC5Q0I pHPT-LoxP-rDNA-mCherry-SKL Peroxisome localization reference plasmid; mCherry-SKL (peroxisome marker) inserted for co-localization verification.
BBa_254B2L3Q Pichia pastoris Das gene Das gene derived from Pichia pastoris, which can increase methanol metabolic flux of Aureobasidium pullulans P16 strain in heterologous expression.
BBa_25D4LV5G pNAT-LoxP-rDNA-P.p Das Overexpression plasmid; Pichia pastoris Das gene inserted to increase Aureobasidium pullulans P16 methanol metabolic flux.
BBa_25Q03R3R pNAT-LoxP-rDNA-yihX Heterologous expression plasmid; Escherichia coli yihX gene inserted to enhance conversion of intermediate products to glucose.
BBa_25YHKFCT Escherichia coli yihX gene yihX gene derived from Escherichia coli, which can enhance the conversion efficiency of metabolic intermediates to glucose during heterologous expression.