Difference between revisions of "Part:BBa K2374002:Design"

(→‎Design Notes)
(→‎Design Notes)
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===Design Notes===
 
===Design Notes===
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We use PCR to clone the GAL80ts from the genomic DNA of ''Saccharomyces cerevisia''S288C.
+
We use PCR to clone the GAL80ts from the genomic DNA of ''Saccharomyces cerevisia'' S288C.
  
 
Then to make 2 site-directed mutagenesis:
 
Then to make 2 site-directed mutagenesis:

Revision as of 03:48, 25 October 2017


GAL80ts (temperature dependent)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 993
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 19
    Illegal BglII site found at 615
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 27
    Illegal BsaI site found at 73


Design Notes

We use PCR to clone the GAL80ts from the genomic DNA of Saccharomyces cerevisia S288C.

Then to make 2 site-directed mutagenesis:

EcoR I (325):GAATTC->GAGTTC There are other mutations during the parts construction by accident: 1142: A->G; aa: Lys->Arg 1154: A->G; aa: Glu->Gly

Source

Chromosome: XIII; NC_001145.3 (171594..172901) NOTE: Gal80ts is a mutation based on the gene above.


References