Difference between revisions of "Part:BBa K2404002:Design"

 
 
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===Design Notes===
 
===Design Notes===
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To isolate this promoter we needed to create a gene construct with the shortened PDF1.2 as the promoter element. We used the Golden Gate cloning system to do this, and so needed to isolate upstream restriction enzyme recognition sites (i.e. Type IIS restriction endonucleases) that are suitable for Golden Gate reactions. This consideration was taken into account when designing primers to isolate this part.
 
  
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- We isolated genomic DNA from wildtype Arabidopsis thaliana
  
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- We designed primers to amplify the PDF1.2 promotor sequence.<br>
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iGEM45 PDF1pF  tttCGTCTCtCTCAggagactacgcgagaacggttcc
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<br>iGEM46 PDF1pR  tttCGTCTCtCTCGcattTGGTAGTGGGTTAATCTTC
  
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- We gel extracted the DNA and digested with BsmBI to introduce the plasmid into pSB1C3.
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- This plasmid corresponds to the [http://2016.igem.org/Resources/Plant_Synthetic_Biology/PhytoBricks Phytobrick] standard.
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===Source===
 
===Source===
  

Latest revision as of 11:27, 6 October 2017


PDF1.2 shortened - a jasmonic acid-induced promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 408
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1
    Illegal BsaI.rc site found at 517


Design Notes

- We isolated genomic DNA from wildtype Arabidopsis thaliana

- We designed primers to amplify the PDF1.2 promotor sequence.
iGEM45 PDF1pF tttCGTCTCtCTCAggagactacgcgagaacggttcc
iGEM46 PDF1pR tttCGTCTCtCTCGcattTGGTAGTGGGTTAATCTTC

- We gel extracted the DNA and digested with BsmBI to introduce the plasmid into pSB1C3.

- This plasmid corresponds to the [http://2016.igem.org/Resources/Plant_Synthetic_Biology/PhytoBricks Phytobrick] standard.


Source

This part comes from Arabidopsis thaliana and was isolated using specific primers.

References