Difference between revisions of "Part:BBa K1510009:Design"
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The promoter, J23100, comes from the 2014 distributed igem kit. The signal sequence, yebF, encoding signal protein exporting recombinant protein extracellular, is from E.coli K-12. By designing primer, we obtain it together with its naturally occurring ribosomal binding site using PCR and extraction. Finally, the terminator B0015 is from 2014 igem distribution. | The promoter, J23100, comes from the 2014 distributed igem kit. The signal sequence, yebF, encoding signal protein exporting recombinant protein extracellular, is from E.coli K-12. By designing primer, we obtain it together with its naturally occurring ribosomal binding site using PCR and extraction. Finally, the terminator B0015 is from 2014 igem distribution. | ||
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[[File:NYMU chienyunghuang yebF KOD.jpg]] | [[File:NYMU chienyunghuang yebF KOD.jpg]] | ||
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===References=== | ===References=== | ||
Latest revision as of 18:18, 17 October 2014
E.coli Signal sequence YebF
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 159
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Similar to BBa_K1190003 from 2013 iGEM Arizona Team, we planned to use YebF to carry protein extracellularly. To fit igem part standard, the circuit is constructed in backbone pSB1C3, chloramphenicol resistant, and with EcoR1 and Xba1 in front of the part, and Spe1, Pst1 behind the part. Sequencing, as well as functional test is done to examine the part.
Source
The promoter, J23100, comes from the 2014 distributed igem kit. The signal sequence, yebF, encoding signal protein exporting recombinant protein extracellular, is from E.coli K-12. By designing primer, we obtain it together with its naturally occurring ribosomal binding site using PCR and extraction. Finally, the terminator B0015 is from 2014 igem distribution.
References
- (2006)"Extracellular accumulation of recombinant proteins fused to the carrier protein YebF in Escherichia coli"
- (2007)"ACES™ Signal Sequence and YebF Expression Systems Technical Brief"
- (2012)"A Protein Export Pathway Involving Escherichia coli Porins"
- (2006)patent" PROTEIN PRODUCTION METHOD UTILIZING YEBF "