Coding

Part:BBa_K3788026

Designed by: Rebecca Pagès   Group: iGEM21_Aix-Marseille   (2021-09-30)


pelB_sfGFP (without stop) _ aida-I

BBa_K3788026 is coding for pelB, sfGFP and AidaA. pelB is a leader sequence that allows the protein to join the periplasm via the SEC system (unfolded proteins). Aida-I, on the other hand, is an autotransporter from the T5SS; it allow the addressing to the extracellular medium. In this contruction sfGFP permit to understand the mechanism, this protein can at the end be replaced by an other protein. sfGFP is supposed to be addressed to the preriplasm thanks to pelB leader sequence and the T5SS system will address sfGFP to the extracellular medium by making it go through a beta-barrel that lead it outside of the cell and then to cleave it. Sequence was optimised for E. coli K12.

T--Aix-Marseille--Figure1_K3788026.jpeg


Figure coming from The Autodisplay Story, from Discovery to Biotechnical and Biomedical Applications

A. representation of the mRNA coding for an autotransporter system.

B. representation of the events of translation, folding in the periplasm and export of the passenger sequence to the extracellular medium.


This figure explains how is constituted a gene coding for an autotransporter such as aidaI. We replaced the signal peptide (SP) by PelB leader sequence and the passenger sequence by sfGFP sequence so that this protein should be transported to the periplasm and then pass to the extracellular medium by the autotransporter AidaI.


Sequence and Features


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 NgoMIV site found at 54
    Illegal AgeI site found at 1864
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 79


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