Difference between revisions of "Part:BBa K2165004:Design"

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===Design Notes===
 
===Design Notes===
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The Yeast Promoter Atlas was used to prevent altering bases that served a significant regulatory purpose (http://ypa.csbb.ntu.edu.tw/do?act=gene_by_kw&query=YHR053C#promoter_map). Base pair number 106 was originally a guanine and was substituted with an adenine in the 5' to 3' direction to eliminate the XbaI site in the middle of the sequence.
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The [http://rulai.cshl.edu/cgi-bin/SCPD/getgene2?CUP1 SCPD] was used to determine whether or not the removal of an internal XbaI site was feasible. <b>Base pair number 84 was originally a guanine and was substituted with an adenine</b> in the 5' to 3' direction to eliminate the XbaI site in the middle of the sequence. This site is one of two Heat Shock Protein binding site, and as such the usage of any assay related to cell stress will likely be less effective with this promoter. A larger Heat Shock Protein binding site remains in tact, but heat shock proteins bind cooperatively (Åkerfelt, 2010).
  
 
===Source===
 
===Source===
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The CUP1 promoter was identified in the 5' untranslated region of the metallothionein gene in Saccharomyces cerevisiae.   
 
The CUP1 promoter was identified in the 5' untranslated region of the metallothionein gene in Saccharomyces cerevisiae.   
  
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===References===
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===Works Cited===
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Åkerfelt, M. Morimoto, R. I. Sistonen, L. Heat shock factors: integrators of cell stress, development and lifespan. Nature Reviews Molecular Cell Biology. 11, 545-555 (2010).

Latest revision as of 02:45, 1 November 2016


CUP1 yeast inducible promoter with RFC[10] restriction sites removed


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]


Design Notes

The [http://rulai.cshl.edu/cgi-bin/SCPD/getgene2?CUP1 SCPD] was used to determine whether or not the removal of an internal XbaI site was feasible. Base pair number 84 was originally a guanine and was substituted with an adenine in the 5' to 3' direction to eliminate the XbaI site in the middle of the sequence. This site is one of two Heat Shock Protein binding site, and as such the usage of any assay related to cell stress will likely be less effective with this promoter. A larger Heat Shock Protein binding site remains in tact, but heat shock proteins bind cooperatively (Åkerfelt, 2010).

Source

The CUP1 promoter was identified in the 5' untranslated region of the metallothionein gene in Saccharomyces cerevisiae.

Works Cited

Åkerfelt, M. Morimoto, R. I. Sistonen, L. Heat shock factors: integrators of cell stress, development and lifespan. Nature Reviews Molecular Cell Biology. 11, 545-555 (2010).