Team:Rice University/CONSTRUCTS

From 2008.igem.org

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         <td>2 Micron ORI and Histadine Tag </td>
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Revision as of 05:23, 29 October 2008


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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
[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

[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]    

 

Novel Zero Leak Inverter Concept (Super Awesome)


Phage Recombination
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