Composite

Part:BBa_K4390059

Designed by: Maarten van den Ancker, Serafina Soehianto   Group: iGEM22_Edinburgh-UHAS_Ghana   (2022-09-03)
Revision as of 15:40, 12 October 2022 by Xianliang (Talk | contribs)


Car9-tagged sfGFP

This part is not compatible with BioBrick RFC10 assembly but is compatible with the iGEM Type IIS Part standard which is also accepted by iGEM.

Usage and Biology

This is one of the three types of silica-binding tags we explored for immobilising metallothioneins to silica beads. Car9 was also a silica-tag part used by the 2021 Edinburgh OG Team.

The green-fluorescent recombinant protein was used as an indicator to show qualitatively and quantitatively whether or not the Car9 peptide has affinity towards silica beads. This was assayed by comparing the green fluorescence intensity between the initial lysate and the supernatant after addition of the silica beads. There should be a significant reduction in fluorescence intensity to prove that the Car9 peptide indeed binds to silica beads and thus, immobilises the sfGFP fused to it.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 11
    Illegal NheI site found at 34
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 145


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Categories
Parameters
None