Team:NEFU China/Basic Part

          

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Basic parts

Number	Name	Description
BBa_K1154001	pFUS1	Our team were aimed at adjusting the expression level of any existing promoter. Thus, we modified the Kozak sequence of the promoter in the plasmid BBa_K775004, a part from the University of the TU-Delft 2012 iGEM team. Our goal is to reduce the expression efficacy of this vector, and make it suitable to express a suicide gene for information destruction. To reduce the expression efficiency of this part in yeast, we modified the Kozak sequence by mutating the sequence “AAAAAA” to “GCCACC”. After that, our information destruction system was built and kill the information host yeast in a desired period of time. See more details in the basic part BBa_K2542014. We used the enhanced green fluorescent protein (EGFP) to test the effect of this modification. The generated plasmid allows a type yeast to sense the α factor secreted by α-type yeast and express EGFP. Thus, when the promoter was activated by  factor secreted by α-type yeast, the EGFP would be expressed. Thus, the EGFP expression would be the standard to tell the strength of our improved promoter.
BBa_K1829002	pFig2c	Our team attempted to verify the strengthen of a promoter called Fig2c in a part of BBa_K1829002 from the iGEM 2015 UCSF team. For this purpose, we inserted the pFig2c promoter and EGFP coding sequence into the pesc-ura plasmid, and transferred it into a-type Saccharomyces cerevisiae. The expression of the Fig2c promoter was induced by adding α mating factor. When the Fig2c promoter is induced, EGFP is expressed. In this way, we can detect the strengthen of the Fig2c promoter using EGFP as a reporter.
BBa_K2542000	Riboswitch1	A type of riboswitch, onstitutive promoter derived from the yeast, and it can initiate medium-intensity gene expression. We use it to control the transcription of our information.
BBa_K2542001	Riboswitch2	A type of riboswitch, onstitutive promoter derived from the yeast, and it can initiate medium-intensity gene expression. We use it to control the transcription of our information.
BBa_K2542002	Riboswitch3	A type of riboswitch, onstitutive promoter derived from the yeast, and it can initiate medium-intensity gene expression. We use it to control the transcription of our information.
BBa_K2542003	Key1	A Small RNA which can change the secondary structure of the riboswitch (BBa_K2542000). It can transcribe a small RNA that adapts to the riboswitch (BBa_K2542000) and alters its secondary structure so that transcription is not blocked by the riboswitch.
BBa_K2542004	Key2	A Small RNA which can change the secondary structure of the riboswitch (BBa_K2542001). It can transcribe a small RNA that adapts to the riboswitch （BBa_K2542001） and alters its secondary structure so that transcription is not blocked by the riboswitch.
BBa_K2542005	Key3	A Small RNA which can change the secondary structure of the riboswitch (BBa_K2542002). It can transcribe a small RNA that adapts to the riboswitch （BBa_K2542002） and alters its secondary structure so that transcription is not blocked by the riboswitch.
BBa_K2542006	pUra	This is a constitutive promoter that initiates gene expression in yeast.
BBa_K2542007	Homologous arm1	It' a part of the transposable element of Saccharomyces cerevisiae, which can be used to perform homologous recombination of DNA to insert the sequence of interest into the genome of Saccharomyces cerevisiae. The other is BBa_K2542008.
BBa_K2542008	Homologous arm2	It' a part of the transposable element of Saccharomyces cerevisiae, which can be used to perform homologous recombination of DNA to insert the sequence of interest into the genome of Saccharomyces cerevisiae. The other is BBa_K2542007.
BBa_K2542009	pRPR1	It' a promoter capable of being recognized by RNA polymerase III, and the RNA driven by it does not add 5' cap and 3' poly A tail. This is a promoter and is usually used with the RPR1 terminator (BBa_K2542010).
BBa_K2542010	tRPR1	This is a terminator and is usually used with the RPR1 promoter (BBa_K2542009).