Difference between revisions of "Part:BBa K1362000:Design"
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===Sources===
 
===Sources===
 
The sources of...
 
The sources of...
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**[Part:BBa_K1362090:Design#source]
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**[[Part:BBa_K1362090:Design|BBa_K1362090 T7 RBS]]
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**<partinfo>BBa_K1362400</partinfo>
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**[[Part:BBa_G0000:Design|BBa_G0000 scar]]
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**<partinfo>BBa_K1362400</partinfo>
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**[[Part:BBa_K1362414:Design|BBa_K1362414 RFC[i] A]]
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**<partinfo>BBa_K1362400</partinfo>
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**[[Part:BBa_K1362401:Design|BBa_K1362401 NpuDnaE(C)]]
−
**<partinfo>BBa_K1362400</partinfo>
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**[[Part:BBa_K1362419:Design|BBa_K1362419 RFC[i] E]]]
−
**<partinfo>BBa_K1362400</partinfo>
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**[[Part:BBa_K1362423:Design|BBa_K1362423 <-BsaI]]
−
**<partinfo>BBa_K1362400</partinfo>
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**[[Part:BBa_J04450:Design|BBa_J04450 RFP coding device]]
−
**<partinfo>BBa_K1362400</partinfo>
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**[[Part:BBa_K1362424:Design|BBa_K1362424 BsaI->]]
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**<partinfo>BBa_K1362400</partinfo>
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**[[Part:BBa_K1362415:Design|BBa_K1362415 RFC[i] A]]
−
 
+
**[[Part:BBa_K1362400:Design|BBa_K1362400 NpuDnaE(N)]]
 +
**[[Part:BBa_K1362421:Design|BBa_K1362421 RFC[i] F]]
 +
**[[Part:BBa_K1362999:Design|BBa_K1362999 iGEMHD tag]]
  
 
===References===
 
===References===

Revision as of 13:21, 17 October 2014

NpuDnaE intein RFC[105] circularization construct


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 931
    Illegal AgeI site found at 1043
  • 1000
    INCOMPATIBLE 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

The sources of...

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).