Difference between revisions of "Part:BBa K3228101:Design"

 
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===Design Notes===
 
===Design Notes===
−
test
 
  
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The crRNA part had to be ordered in 4 different parts, because there are direct repeats in between the different gRNA spaceholders. These 4 parts were ordered with BsaI overhangs that were cut out with a Golden Gate reaction and the final construct was assembled via annealing of the resulting overhangs. We use a GFP cassette as a placeholder for the gRNA, that can be cut out via BsmbI digest. The part reaches from the beginning of the RBS overhang to the end of the Terminator.
  
  
 
===Source===
 
===Source===
  
−
test
+
The crRNA was synthesized in 4 parts via IDT gene synthesis.
  
 
===References===
 
===References===

Revision as of 16:54, 18 October 2019


pMC_0_3-5_1_crRNA-GFP_dropout


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1186
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 1186
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 1186
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 242


Design Notes

The crRNA part had to be ordered in 4 different parts, because there are direct repeats in between the different gRNA spaceholders. These 4 parts were ordered with BsaI overhangs that were cut out with a Golden Gate reaction and the final construct was assembled via annealing of the resulting overhangs. We use a GFP cassette as a placeholder for the gRNA, that can be cut out via BsmbI digest. The part reaches from the beginning of the RBS overhang to the end of the Terminator.


Source

The crRNA was synthesized in 4 parts via IDT gene synthesis.

References