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. | ||
| + | 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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Revision as of 10:19, 27 September 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.
Experiment 1:
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.
- yeast-sfGFP-ADH1-Kan ( https://parts.igem.org/Part:BBa_K1486026)
- yeast-sfGFP-N ( https://parts.igem.org/Part:BBa_K1486028)
- yeast-sfGFP_N-ADH1-Kan ( https://parts.igem.org/Part:BBa_K1486029)
- yeast-sfGFP-ADH1-Ura ( https://parts.igem.org/Part:BBa_K1486032)
- yeast-sfGFP-C( https://parts.igem.org/Part:BBa_K1486034)
- yeast-sfGFP_C-ADH1-Ura( https://parts.igem.org/Part:BBa_K1486035)
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
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 2213
Illegal PstI site found at 3446 - 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1205
Illegal PstI site found at 2213
Illegal PstI site found at 3446 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 3657
Illegal BamHI site found at 1144
Illegal XhoI site found at 1283
Illegal XhoI site found at 2470 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 2213
Illegal PstI site found at 3446 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 2213
Illegal PstI site found at 3446
Illegal AgeI site found at 979
Illegal AgeI site found at 1694
Illegal AgeI site found at 2881 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 961