Difference between revisions of "Part:BBa K1362000:Design"
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*[[Part:BBa_K1362090:Design|BBa_K1362090 T7 RBS]] | *[[Part:BBa_K1362090:Design|BBa_K1362090 T7 RBS]] | ||
*[[Part:BBa_G0000:Design|BBa_G0000 scar]] | *[[Part:BBa_G0000:Design|BBa_G0000 scar]] | ||
| − | *[[Part:BBa_K1362414:Design|BBa_K1362414 RFC[ | + | *[[Part:BBa_K1362414:Design|BBa_K1362414 RFC[105] A]] |
*[[Part:BBa_K1362401:Design|BBa_K1362401 NpuDnaE(C)]] | *[[Part:BBa_K1362401:Design|BBa_K1362401 NpuDnaE(C)]] | ||
| − | *[[Part:BBa_K1362419:Design|BBa_K1362419 RFC[ | + | *[[Part:BBa_K1362419:Design|BBa_K1362419 RFC[105] E]]] |
*[[Part:BBa_K1362423:Design|BBa_K1362423 <-BsaI]] | *[[Part:BBa_K1362423:Design|BBa_K1362423 <-BsaI]] | ||
*[[Part:BBa_J04450:Design|BBa_J04450 RFP coding device]] | *[[Part:BBa_J04450:Design|BBa_J04450 RFP coding device]] | ||
*[[Part:BBa_K1362424:Design|BBa_K1362424 BsaI->]] | *[[Part:BBa_K1362424:Design|BBa_K1362424 BsaI->]] | ||
| − | *[[Part:BBa_K1362415:Design|BBa_K1362415 RFC[ | + | *[[Part:BBa_K1362415:Design|BBa_K1362415 RFC[105] A]] |
*[[Part:BBa_K1362400:Design|BBa_K1362400 NpuDnaE(N)]] | *[[Part:BBa_K1362400:Design|BBa_K1362400 NpuDnaE(N)]] | ||
| − | *[[Part:BBa_K1362421:Design|BBa_K1362421 RFC[ | + | *[[Part:BBa_K1362421:Design|BBa_K1362421 RFC[105] F]] |
*[[Part:BBa_K1362999:Design|BBa_K1362999 iGEMHD tag]] | *[[Part:BBa_K1362999:Design|BBa_K1362999 iGEMHD tag]] | ||
Latest revision as of 15:57, 24 October 2014
NpuDnaE intein RFC[105] circularization construct
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 931
Illegal AgeI site found at 1043 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1220
Illegal BsaI.rc site found at 145
Design Notes
Between the coding sequences of the Npu DnaE C-intein BBa_K1362401 and the N-intein BBa_K1362400 we placed BBa_J04450, an mRFP selection marker flanked by BsaI sites that can be replaced by a protein of interest. A strong RBS BBa_K1362090 was added. This part was assembled by CPEC [1] from PCR products of BBa_K1362093 , BBa_J04450, pVS07 and pVS41 [2].
Sources of the Subparts
- BBa_K1362090 T7 RBS
- BBa_G0000 scar
- BBa_K1362414 RFC[105] A
- BBa_K1362401 NpuDnaE(C)
- BBa_K1362419 RFC[105] E]
- BBa_K1362423 <-BsaI
- BBa_J04450 RFP coding device
- BBa_K1362424 BsaI->
- BBa_K1362415 RFC[105] A
- BBa_K1362400 NpuDnaE(N)
- BBa_K1362421 RFC[105] F
- BBa_K1362999 iGEMHD tag
References
[1] Quan, J. & Tian, J. Circular polymerase extension cloning for high-throughput cloning of complex and combinatorial DNA libraries. Nat. Protoc. 6, 242–51 (2011).
[2] Zettler, J., Schütz, V. & Mootz, H. D. The naturally split Npu DnaE intein exhibits an extraordinarily high rate in the protein trans-splicing reaction. FEBS Lett. 583, 909–14 (2009).