Coding

Part:BBa_K1510010:Design

Designed by: Chien-Yung Huang   Group: iGEM14_NYMU-Taipei   (2014-10-06)

Extracellular RFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 161
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 965
    Illegal AgeI site found at 1077
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

In order to make red florescent protein(RFP), being secreted as recombinant protein with yebF, we design primer with special prefix and suffix(RFC23) between yebF and RFP in order to make this recombinant protein. At the end, we put it into pSB1C3, so it is standard biobrick compatible with EcoR1, Xba1, Spe1, and Pst1 cutting sites.


Source

The strong constitutive promoter, J23100 is from 2014 igem tool kit. At the down stream of promoter, yebF is a signal sequence, together with its own ribosomal binding site, from E.coli K12. After yebF, the red florescent sequence from igem 2014 tool kit is used. Finally, a commonly used igem from 2014 distribution double terminator, B0015, is constructed.

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 "