Plasmid

Part:BBa_K5173005

Designed by: Gaocheng Zeng   Group: iGEM24_EHSBNU   (2024-10-02)


Pfastbac1 plasmid with optimized tachypleus tridentatus clotting factor G beta subunit

Since prokaryotic expression hosts may lack the protein modification mechanisms required for producing a fully functional horseshoe crab protein, we also conducted experiments with Spodoptera frugiperda 9 insect cells. We utilized the Bac-to-Bac expression system, where this plasmid (Pfastbac1) serves as a transfer plasmid for the beta subunit. Within the insect baculovirus expression system, transposition of the gene of interest to the Bacmid's attTn7 site is facilitated by a helper plasmid providing trans-acting functions. By employing this method, we are able to obtain recombinant viruses with a 100% positive rate.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2547
    Illegal BglII site found at 3017
    Illegal BamHI site found at 4032
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 127
  • 1000
    COMPATIBLE WITH RFC[1000]


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