Difference between revisions of "Part:BBa K1638011:Design"
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===References=== | ===References=== | ||
| + | [1]: Karimova G, Pidoux J, Ullmann A, Ladant D. A bacterial two-hybrid system based on a reconstituted signal transduction pathway. Proceedings of the National Academy of Sciences of the United States of America. 1998;95(10):5752-6. | ||
Latest revision as of 09:21, 18 September 2015
Leucine zipper fused to T18 domain of cyaA from Bordetella pertussis
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 711
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 252
Illegal NgoMIV site found at 662
Illegal AgeI site found at 468 - 1000COMPATIBLE 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 and BBa_C2001
References
[1]: Karimova G, Pidoux J, Ullmann A, Ladant D. A bacterial two-hybrid system based on a reconstituted signal transduction pathway. Proceedings of the National Academy of Sciences of the United States of America. 1998;95(10):5752-6.