Composite

Part:BBa_K3739112

Designed by: Shichen Geng   Group: iGEM21_XMU-China   (2021-09-30)
Revision as of 00:45, 22 October 2021 by Master123 (Talk | contribs)

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K525998(T7-RBS)-pelB-Ag43-160N-his-GFP-B0010

Ag43 is a surface display protein and GFP can show the expression effciency of Ag43

Usage and Biology

Ag43 is a surface display protein that displays protein molecule it carries on the cell membrane of bacteria. pelB is an exopeptide linked to Ag43. Tevp can shear Ag43 and release the exopeptide pelB to secrete the protein molecules out of bacteria.

We used BBa_J23100 to construct the expression system and obtained the composite BBa_K3739058, which is assembled on the expression vector pET-28(+) by standard assembly. The constructed plasmid was transformed into Vibrio natriegens , then the positive transformants were selected by kanamycin and confirmed by colony PCR and sequencing.

Reference

1. Ahan, R. E.; Kirpat, B. M.; Saltepe, B.; Seker, U. O. S., A Self-Actuated Cellular Protein Delivery Machine. ACS Synth Biol 2019, 8 (4), 686-696.


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 92
    Illegal NgoMIV site found at 1050
    Illegal NgoMIV site found at 2274
    Illegal NgoMIV site found at 2784
    Illegal NgoMIV site found at 2805
    Illegal AgeI site found at 1800
    Illegal AgeI site found at 2544
    Illegal AgeI site found at 2958
    Illegal AgeI site found at 3200
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 180


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