Difference between revisions of "Part:BBa K1638031:Design"

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===Source===
 
===Source===
−
pUT18C and <partinfo>BBa_C2001</partinfo>
+
pUT18C, <partinfo>BBa_C2001</partinfo> and <partinfo>BBa_K861173</partinfo>
  
 
===References===
 
===References===

Revision as of 09:19, 18 September 2015


Leucine zipper fused to T18 domain of cyaA with cAMP-induced RFP generator


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1719
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1260
    Illegal NgoMIV site found at 1670
    Illegal AgeI site found at 712
    Illegal AgeI site found at 824
    Illegal AgeI site found at 1476
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part was first constructed by amplifying the leucine zipper region from BBa_C2001 using the primers:

- Forward: 5’-CGCTTCTAGAGGGATCCGAAAATTTGTATTTTCAATCTGGTATGAAACAGCTGGAAGACAAAGTTGA-3’ (BamHI res. site included) - Reverse: 5’-ATATCTGCAGCGGCCGCTACTAGTAACGTTCACCAACCAGTTTTTTCAGA-3’

By digesting the amplified leucine zipper product and the backbone containing either the T25 or T18 domain of the adenylate cyclase cyaA from Bordetella pertussis with BamHI and PstI, we were able to ligate the two bricks together without creating a scar-site.

Source

pUT18C, BBa_C2001 and BBa_K861173

References