Difference between revisions of "Part:BBa K2404005:Design"

 
 
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===Design Notes===
 
===Design Notes===
 +
- We isolated genomic DNA from wildtype Arabidopsis thaliana
 +
 +
- We designed primers to amplify the WRKY30 promotor sequence.<br>
 +
iGEM_59 WRKY30p2F tttCGTCTCtCTCAcgcctagattgtctaaacgtcg
 +
<br>iGEM_67 WRKY30p2Ra tttCGTCTCtCTCGcattGGTTTGTGAGAAGTCAGACCTCTTC
 +
 +
- 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.
 +
 +
 +
 
To isolate this promoter we needed to create a gene construct with the WRKY30 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.
 
To isolate this promoter we needed to create a gene construct with the WRKY30 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.
  

Latest revision as of 12:04, 6 October 2017


WRKY30 - a promoter induced by the presence of cellulose-derived oligomers


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 122
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 122
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 122
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1
    Illegal BsaI site found at 477
    Illegal BsaI.rc site found at 502


Design Notes

- We isolated genomic DNA from wildtype Arabidopsis thaliana

- We designed primers to amplify the WRKY30 promotor sequence.
iGEM_59 WRKY30p2F tttCGTCTCtCTCAcgcctagattgtctaaacgtcg
iGEM_67 WRKY30p2Ra tttCGTCTCtCTCGcattGGTTTGTGAGAAGTCAGACCTCTTC

- 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.


To isolate this promoter we needed to create a gene construct with the WRKY30 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.


Source

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

References