Team:Rice University/CONSTRUCTS

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Revision as of 00:42, 30 October 2008

OUR TEAM:::SUMMARY ::: BACKGROUND ::: STRATEGY ::: RESULTS ::: ONGOING WORK :::

Saccharomyces cerevisiae are widely used for baking and brewing, and they are particular useful for synthesizing metabolites under fermentation conditions which prevent the air oxidation of many useful compounds. To achieve our project and expand the synthetic biology toolbox for programming yeast, we have introduced into the iGem registry BioBricks encoding 3 yeast promoters, 3 yeast terminators, a two micron origin of replication, 2 selectable markers, 2 enzymes, and a yeast integration plasmid. In addition, we have generated seven constructs using these parts. Furthermore, we have submitted two additional parts representing a foundational tool, including a gene encoding an amber suppressed RFP biobrick for screening of SupF+ (Amber suppressor) genotype and an amber suppressor tRNA biobrick.

Yeast Regulator Biobricks

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

Yeast Promoters
BBa_K122000	pPGK1	This is the 1500 bp upstream of the PGK1 coding region in an industrial yeast strain. Constitutive promoter.	1497
BBa_K122002	pADH1	700bp upstream of ADH1 promoter region containing RBS. Constitutive promoter.	701
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
BBa_K122003	tCYC1	300bp downstream the CYC1 coding region in a standard yeast strain.	300
BBa_K122004	tADH1	300bp downstream the ADH1 coding region in a standard yeast strain.	300
BBa_K122013	tPGK1	1000bp downstream the PGK1 coding region in an industrial yeast strain.	1000

Selectable Markers

Selectable Markers
BBa_K122018	ZeoR	Zeocin Resistance Gene	300
BBa_K122008	BleoR	Bleocin Resistance Gene under pTet promoter	800ish
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.

BBa_K122001	[pGAL1][tetO][ZeoR]
BBa_K122005	Tyrosine Ammonia Lyase
BBa_K122010	4CL:STS
BBa_K122012	[pPGK1][4CL:STS][tCYC1]
BBa_K122015	[pGAL1][tetO][ZeoR][tADH1]
BBa_K122021	[pADH1][TAL][tPGK1]
BBa_K122019	[pPGK1][4CL:STS][tCYC1][pGAL1][tetO][ZeoR][tADH1]

Additional Bacterial Parts

Novel Zero Leak Inverter Concept

wtRFP in SupF- / ARFP in SupF- /wtRFP in SupF+ / ARFP in SupF+

@@ Line 130: / Line 130: @@
        </tr>
        <tr>
-         <td>[http://partsregistry.org/wiki/index.php?title=Part:BBa_K122016 BBa_K122016]</td>
+         <td>[http://partsregistry.org/wiki/index.php?title=Part:BBa_K122021 BBa_K122021]</td>
-         <td>4CL:STS with CYC1 terminator.</td>
+         <td>[pADH1][TAL][tPGK1]</td>
          <td>[[Image:C3.jpg|150px]]</td>
          <td>&nbsp;</td>