Constructs

Saccharomyces cerevisiae is a versatile eukaryotic model organism studied for its fermentive ability. The conditions of fermentation (low oxygen content and cool, dark environment) are uniquely suitable for the synthesis and transport antioxidant phytophlactics. Synthetic biology work using the yeast model system is currently limited by the number of yeast biobrick parts available in the registry. In addition to introducing novel genes into the registry, this year we added 3 yeast promoters, 3 yeast terminators, a 2 micron origin of replication, two selectable markers, and a yeast integration plasmid suitable for future yeast work. For additional foundational tools, we have added an amber suppressed RFP biobrick for screening of SupF+ (Amber suppressor) genotype with an amber suppressor tRNA biobrick.

Yeast Regulator Biobricks

Three unique constitutive and repressible yeast promoters were introduced into the registry.

Yeast Promoters
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K122000 BBa_K122000]	pPGK1	This is the 1500 bp upstream of the PGK1 coding region in an industrial yeast strain. Constitutive promoter.	1497
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K122002 BBa_K122002]	pADH1	700bp upstream of ADH1 promoter region containing RBS. Constitutive promoter.	701
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K122001 BBa_K122017]	pGAL1 + tetO	The GAL1 promoter which is highly repressed by glucose. An additional tetracycline operator site was included upstream of the RBS to allow repression by tetR.	484

Four additional yeast terminators were introduced into the registry.

Yeast Terminators
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K122003 BBa_K122003]	tCYC1	300bp downstream the CYC1 coding region in a standard yeast strain.	300
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K122004 BBa_K122004]	tADH1	300bp downstream the ADH1 coding region in a standard yeast strain.	300
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K122013 BBa_K122013]	tPGK1	1000bp downstream the PGK1 coding region in an industrial yeast strain.	1000

Selectable Markers

Selectable Markers
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K122001 BBa_K122018]	ZeoR	Zeocin Resistance Gene	300
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K122008 BBa_K122008]	BleoR	Bleocin Resistance Gene under pTet promoter	800ish
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K122014 BBa_K122014]	ORI+HisTag	2 Micron ORI and Histadine Tag	<9000

Project Specific Constructs

This year's project involved the integration of 4-coumarate:coenzyme A ligase/ stilbene synthase fusion protein, a tyrosine ammonia lyase, and a zeocin resistance selectable marker. For more information, please visit Strategy.

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K122001 BBa_K122001]	[pGAL1][tetO][ZeoR]
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K122005 BBa_K122005]	Tyrosine Ammonia Lyase
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K122010 BBa_K122010]	4CL:STS
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K122012 BBa_K122012]	[pPGK1][4CL:STS][tCYC1]
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K122015 BBa_K122015]	[pGAL1][tetO][ZeoR][tADH1]
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K122016 BBa_K122016]	4CL:STS with CYC1 terminator.
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K122016 BBa_K122019]	[pPGK1][4CL:STS][tCYC1][pGAL1][tetO][ZeoR][tADH1]