Difference between revisions of "Part:BBa K2404004:Design"

 
 
Line 7: Line 7:
  
 
===Design Notes===
 
===Design Notes===
−
To isolate this promoter we needed to create a gene construct with the shortened GST6 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.
+
- We isolated genomic DNA from wildtype Arabidopsis thaliana
  
 +
- We designed primers to amplify the GST6 promotor sequence.<br>
 +
iGEM49 GST6pF tttCGTCTCtCTCAggagGGAAAGATGAGATTCGTTTC
 +
<br>iGEM50 GST6pR tttCGTCTCtCTCGcattGGAATTGGATGAAGAAGAATG
 +
 +
- 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.
  
  

Latest revision as of 11:45, 6 October 2017


GST6 shortened - a salicylic acid-induced promoter


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 68
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 68
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 68
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1
    Illegal BsaI.rc site found at 514


Design Notes

- We isolated genomic DNA from wildtype Arabidopsis thaliana

- We designed primers to amplify the GST6 promotor sequence.
iGEM49 GST6pF tttCGTCTCtCTCAggagGGAAAGATGAGATTCGTTTC
iGEM50 GST6pR tttCGTCTCtCTCGcattGGAATTGGATGAAGAAGAATG

- 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