Difference between revisions of "Part:BBa K3373014"
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<partinfo>BBa_K3373014 short</partinfo> | <partinfo>BBa_K3373014 short</partinfo> | ||
− | a | + | 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. |
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Latest revision as of 12:10, 25 October 2020
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
- 10INCOMPATIBLE 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 - 12INCOMPATIBLE 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 - 21INCOMPATIBLE 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 - 23INCOMPATIBLE 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 - 25INCOMPATIBLE 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 - 1000INCOMPATIBLE 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