Composite

Part:BBa_K4621131:Design

Designed by: Anyang Chen, Ruikang Chen   Group: iGEM23_SCUT-China   (2023-10-05)


Complex functional parts of SCUT-3-EctPi4 that induce high Hydroxyectoine production.


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 3756
    Illegal SpeI site found at 8275
    Illegal PstI site found at 3437
    Illegal PstI site found at 3744
    Illegal PstI site found at 6169
    Illegal PstI site found at 6403
    Illegal PstI site found at 7615
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 8252
    Illegal SpeI site found at 8275
    Illegal PstI site found at 3437
    Illegal PstI site found at 3744
    Illegal PstI site found at 6169
    Illegal PstI site found at 6403
    Illegal PstI site found at 7615
    Illegal NotI site found at 781
    Illegal NotI site found at 1666
    Illegal NotI site found at 1984
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 4208
    Illegal BglII site found at 5003
    Illegal BglII site found at 7772
    Illegal BamHI site found at 3948
    Illegal BamHI site found at 5297
    Illegal BamHI site found at 8240
    Illegal BamHI site found at 8445
    Illegal XhoI site found at 963
    Illegal XhoI site found at 1137
    Illegal XhoI site found at 4621
    Illegal XhoI site found at 6967
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 3756
    Illegal SpeI site found at 8275
    Illegal PstI site found at 3437
    Illegal PstI site found at 3744
    Illegal PstI site found at 6169
    Illegal PstI site found at 6403
    Illegal PstI site found at 7615
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 3756
    Illegal SpeI site found at 8275
    Illegal PstI site found at 3437
    Illegal PstI site found at 3744
    Illegal PstI site found at 6169
    Illegal PstI site found at 6403
    Illegal PstI site found at 7615
    Illegal NgoMIV site found at 998
    Illegal NgoMIV site found at 1404
    Illegal NgoMIV site found at 1410
    Illegal NgoMIV site found at 1701
    Illegal NgoMIV site found at 2750
    Illegal NgoMIV site found at 2942
    Illegal NgoMIV site found at 3452
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 586
    Illegal BsaI.rc site found at 766
    Illegal BsaI.rc site found at 1548
    Illegal BsaI.rc site found at 2076
    Illegal BsaI.rc site found at 2331
    Illegal BsaI.rc site found at 2421
    Illegal BsaI.rc site found at 2496
    Illegal SapI site found at 624
    Illegal SapI site found at 4539
    Illegal SapI site found at 5829
    Illegal SapI.rc site found at 7026


Design Notes

The effectiveness, and GC content.


Source

The CRISPRi fragment is derived from PZ-ProP-Ect-i4 plasmid, then be ligated after EctOperon on the PZ-ProP-Ect-i4 plasmid.

References

[1] Zhao Y, Li L, Zheng G, Jiang W, Deng Z, Wang Z, Lu Y. CRISPR/dCas9-Mediated Multiplex Gene Repression in Streptomyces. Biotechnol J. 2018 Sep;13(9):e1800121. doi: 10.1002/biot.201800121. Epub 2018 Jul 4. PMID: 29862648.