Basic Parts
BBa_K3506007 is the promoter of the G2/mitotic-specific Cyclin-B2 encoding gene (CLB2) which initiates its transcription in late S and G2/M in the yeast cell cycle. This part is used to control the periodic expression of Cas9 so that we can couple the expression of Cas9 to cell division.
BBa_K3506010 is the first 124 amino acids of Cyclin B2 in Saccharomyces cerevisiae. It functions as a degradation tag and promotes the degradation of Clb2 protein.
BBa_K3506021 is an RNA polymerase III promoter and is a common part of eukaryotic expression systems. We use it to initiate the transcription of hgRNA.
BBa_K3506050 is a single guide RNA that guides Cas9 to target ADE2 gene in Cryptococcus neoformans.
Composite Parts
The cassette of periodic expression of Cas9
The cassette of periodic expression and degradation of Cas9
GFP with degradation tag Clb2 N124aa
Constitutive double promoter module
Inducible double promoter module
BBa_K3506009 consists of two parts: CLB2 promoter (BBa_K3506007) and Cas9 (BBa_K2130013). This part enables the CLB2 promoter to initiate the expression of Cas9, so as to achieve the goal of coupling the expression of Cas9 with the cell cycle.
BBa_K3506030 consists of four parts: CLB2 promoter (BBa_K3506007), the first 124 amino acids of Clb2 (BBa_K3506010), a linker (BBa_K3506008) and Cas9 (BBa_K2130013). CLB2 promoter initiates the expression of Cas9 at S phase. The first 124 amino acids of Clb2 degrades Cas9 quickly. By this way, we control the expression and degradation of Cas9 periodically.
BBa_K3506036 consists of three parts: Clb2 124aa (BBa_K3506010) and GFP (BBa_E0040) through Linker (BBa_K3506013). It is designed to characterize the degradation rate of Clb2 N124aa.
BBa_K3506095 consists of two parts: GAPDH promoter (BBa_K3506099) and U6 promoter (BBa_K3506021). It is used to read the lineage tracing information at RNA level at specific time.
BBa_K3506022 consists of two parts: GAL7 promoter (BBa_K3506424) and U6 promoter (BBa_K3506021). It is used to read the lineage tracing information at RNA level at specific time.
