Difference between revisions of "Part:BBa K1486023"
 
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This part corresponds to the sequence of the superfolded GFP optimized for yeast cells. We used it as a tag to control the expression of our genes of interest Hog1 and Pbs2. It was extracted by PCR from plasmids we ordered to addgene:Plasmid 44901 pFA6a-link-yoSuperfolderGFP-Kan and Plasmid 44873 pFA6a-link-yoSuperfolderGFP-CaURA3.  
 
This part corresponds to the sequence of the superfolded GFP optimized for yeast cells. We used it as a tag to control the expression of our genes of interest Hog1 and Pbs2. It was extracted by PCR from plasmids we ordered to addgene:Plasmid 44901 pFA6a-link-yoSuperfolderGFP-Kan and Plasmid 44873 pFA6a-link-yoSuperfolderGFP-CaURA3.  
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This construct was also used to evaluate the interaction between hog1 and pbs2 in S.Cerevisae: we split the sequence and fused the N-Terminal part to pbs2 and the C-Terminal part to hog1.
 
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===Experiment 1: CpxR dimerization & Dimerization Orientation===
 
 
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===Associated Biobricks===
 
===Associated Biobricks===
 
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<FONT SIZE="2"><P>In the context of the same experiment, we designed three more constructs with different CpxR - IFP fragment orientations. This construct was made in all orientations: 
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<FONT SIZE="2"><P>We assembled this part with the yeast resistance genes and the terminator ADH1. We also made a split sfGFP to evaluate the dimerization of hog1 and pbs2 in response to osmotic stress.
 
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<LI> CxpR & Split IFP1.4 [Nterm + Nterm] ( https://parts.igem.org/Part:BBa_K1486009 )
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<LI> yeast-sfGFP-ADH1-Kan ( https://parts.igem.org/Part:BBa_K1486026)
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<LI> CxpR & Split IFP1.4 [Nterm + Cterm] ( https://parts.igem.org/Part:BBa_K1486010 )
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<LI> yeast-sfGFP-N ( https://parts.igem.org/Part:BBa_K1486028)
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<LI> CxpR & Split IFP1.4 [Cterm + Nterm] ( https://parts.igem.org/Part:BBa_K1486011 )
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<LI> yeast-sfGFP_N-ADH1-Kan ( https://parts.igem.org/Part:BBa_K1486029)
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<LI> yeast-sfGFP-ADH1-Ura ( https://parts.igem.org/Part:BBa_K1486032)
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<LI> yeast-sfGFP-C( https://parts.igem.org/Part:BBa_K1486034)
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<LI> yeast-sfGFP_C-ADH1-Ura( https://parts.igem.org/Part:BBa_K1486035)
 
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===References===
 
===References===
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WARNING: This Biobrick was built by Gibson Assembly and contains illegal restriction restriction enzyme sites !
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K1486008 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K1486023 SequenceAndFeatures</partinfo>
  
  
 
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K1486008 parameters</partinfo>
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<partinfo>BBa_K1486023 parameters</partinfo>
 
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Latest revision as of 16:31, 14 October 2014

Yeast optimized superfolder GFP

Purpose of the Biobrick

This part corresponds to the sequence of the superfolded GFP optimized for yeast cells. We used it as a tag to control the expression of our genes of interest Hog1 and Pbs2. It was extracted by PCR from plasmids we ordered to addgene:Plasmid 44901 pFA6a-link-yoSuperfolderGFP-Kan and Plasmid 44873 pFA6a-link-yoSuperfolderGFP-CaURA3. This construct was also used to evaluate the interaction between hog1 and pbs2 in S.Cerevisae: we split the sequence and fused the N-Terminal part to pbs2 and the C-Terminal part to hog1.

Associated Biobricks

We assembled this part with the yeast resistance genes and the terminator ADH1. We also made a split sfGFP to evaluate the dimerization of hog1 and pbs2 in response to osmotic stress.

References

Improved Blue, Green, and Red Fluorescent Protein Tagging Vectors for S. cerevisiae. Lee et al (PLoS One. 2013 Jul 2;8(7):e67902. doi: 10.1371/journal.pone.0067902. Print 2013. PubMed)



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]