Team:Rice University/CONSTRUCTS

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SUMMARY ::: BACKGROUND ::: STRATEGY ::: CONSTRUCTS ::: RESULTS ::: ONGOING WORK ::: OUR TEAM ::: NOTEBOOK ::: GALLERY

Contents

Constructs

S. cerevisiae is an incredible eukaryotic model organism, but its potential for synthetic biology is hidden by the lack of modular biobrick parts for this system. In addition to introducing novel genes into the registry, this year we added new yeast promoters, yeast terminators, a Histidine tag and 2 micron ORI, two new selectible markers, and a yeast integration plasmid suitable for future yeast work. For additional foundational tools, we have added an amber suppressor biobrick with a wide range of functionality and selectible by the amber suppressed RFP.

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
<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K122001">BBa_K122017</a> 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
<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K122003">BBa_K122003</a> tCYC1 300bp downstream the CYC1 coding region in a standard yeast strain. 300
<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K122004">BBa_K122004</a> tADH1 300bp downstream the ADH1 coding region in a standard yeast strain. 300
<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K122013">BBa_K122013</a> tPGK1 1000bp downstream the PGK1 coding region in an industrial yeast strain. 1000

 

Selectable Markers

Selectable Markers
<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K122001">BBa_K122018</a><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K122003"></a> ZeoR Zeocin Resistance Gene 300
<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K122008">BBa_K122008</a><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K122004"></a> BleoR Bleocin Resistance Gene under pTet promoter 800ish
<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K122014">BBa_K122014</a><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K122013"></a> ORI+HisTag 2 Micron ORI and Histadine Tag <9000

 

Project Specific Constructs

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

 

Novel Zero Leak Inverter Concept (Super Awesome)


Phage Recombination