Composite

Part:BBa_K3373014

Designed by: Antonin Chevenier   Group: iGEM20_Sorbonne_U_Paris   (2020-10-23)


BSR_NucA-TEVsite_UVR8-NtTEV_COP1-CtTEV

This part is a multigenic construct containing our three constructs for the operation of our kill-switch system adapted to C. reinhardtii. The CDS of the Nuclease A (NucA) fused to a double nuclear localization signal SV40 NLS (BBa_K3373008) is associated at the N-terminus with the HAP2 transmembrane domain (BBa_K3373007) and at the C-terminus with the recognition site for the TEV protease associated with a peptide linker (BBa_K3373011). The CDS of UVR8 (BBa_K3373005) is fused to the C-terminus domain of the TEV protease (BBa_K3373010) with the GSAT peptide linker (BBa_K3373006). The CDS of COP1 (BBa_K3373004) is fused to the N-terminus domain of the TEV protease (BBa_K3373009) with the GSAT peptide linker (BBa_K3373006). All these CDS are under the control of the PSAD promoter and terminator which allow a strong expression in C. reinhardtii. It also contains a Blasticidin resistance cassette (BSR) encoding for the Blasticidin S Deaminase in order to select the recombinant clones after transformation in C. reinhardtii.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1
    Illegal EcoRI site found at 1356
    Illegal EcoRI site found at 3476
    Illegal EcoRI site found at 6675
    Illegal PstI site found at 4323
    Illegal PstI site found at 7893
    Illegal PstI site found at 8070
    Illegal PstI site found at 8418
    Illegal PstI site found at 8433
    Illegal PstI site found at 9858
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1
    Illegal EcoRI site found at 1356
    Illegal EcoRI site found at 3476
    Illegal EcoRI site found at 6675
    Illegal NheI site found at 460
    Illegal PstI site found at 4323
    Illegal PstI site found at 7893
    Illegal PstI site found at 8070
    Illegal PstI site found at 8418
    Illegal PstI site found at 8433
    Illegal PstI site found at 9858
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1
    Illegal EcoRI site found at 1356
    Illegal EcoRI site found at 3476
    Illegal EcoRI site found at 6675
    Illegal BglII site found at 5145
    Illegal BamHI site found at 7
    Illegal BamHI site found at 5814
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1
    Illegal EcoRI site found at 1356
    Illegal EcoRI site found at 3476
    Illegal EcoRI site found at 6675
    Illegal PstI site found at 4323
    Illegal PstI site found at 7893
    Illegal PstI site found at 8070
    Illegal PstI site found at 8418
    Illegal PstI site found at 8433
    Illegal PstI site found at 9858
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1
    Illegal EcoRI site found at 1356
    Illegal EcoRI site found at 3476
    Illegal EcoRI site found at 6675
    Illegal PstI site found at 4323
    Illegal PstI site found at 7893
    Illegal PstI site found at 8070
    Illegal PstI site found at 8418
    Illegal PstI site found at 8433
    Illegal PstI site found at 9858
    Illegal NgoMIV site found at 4370
    Illegal NgoMIV site found at 5639
    Illegal NgoMIV site found at 5699
    Illegal NgoMIV site found at 8723
    Illegal NgoMIV site found at 9429
    Illegal NgoMIV site found at 9543
    Illegal NgoMIV site found at 9603
    Illegal AgeI site found at 4967
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 10586
    Illegal SapI.rc site found at 2947
    Illegal SapI.rc site found at 5740
    Illegal SapI.rc site found at 6146
    Illegal SapI.rc site found at 10032


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