Coding

Part:BBa_K3739022

Designed by: Han Ziyan   Group: iGEM21_XMU-China   (2021-09-08)
Revision as of 05:02, 21 October 2021 by Ettrium (Talk | contribs)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)


pelB-Ag43-160N-his-GFP

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 54
    Illegal NgoMIV site found at 1012
    Illegal NgoMIV site found at 2236
    Illegal NgoMIV site found at 2746
    Illegal NgoMIV site found at 2767
    Illegal AgeI site found at 1762
    Illegal AgeI site found at 2506
    Illegal AgeI site found at 2920
    Illegal AgeI site found at 3162
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 142


[edit]
Categories
Parameters
None