Composite

Part:BBa_K4621130:Design

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


Complex functional parts of SCUT-3-EctPi2 that induce high Hydroxyectoine production.


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 3741
    Illegal SpeI site found at 8260
    Illegal PstI site found at 3437
    Illegal PstI site found at 3729
    Illegal PstI site found at 6154
    Illegal PstI site found at 6388
    Illegal PstI site found at 7600
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 8237
    Illegal SpeI site found at 8260
    Illegal PstI site found at 3437
    Illegal PstI site found at 3729
    Illegal PstI site found at 6154
    Illegal PstI site found at 6388
    Illegal PstI site found at 7600
    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 4193
    Illegal BglII site found at 4988
    Illegal BglII site found at 7757
    Illegal BamHI site found at 3933
    Illegal BamHI site found at 5282
    Illegal BamHI site found at 8225
    Illegal XhoI site found at 963
    Illegal XhoI site found at 1137
    Illegal XhoI site found at 4606
    Illegal XhoI site found at 6952
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 3741
    Illegal SpeI site found at 8260
    Illegal PstI site found at 3437
    Illegal PstI site found at 3729
    Illegal PstI site found at 6154
    Illegal PstI site found at 6388
    Illegal PstI site found at 7600
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 3741
    Illegal SpeI site found at 8260
    Illegal PstI site found at 3437
    Illegal PstI site found at 3729
    Illegal PstI site found at 6154
    Illegal PstI site found at 6388
    Illegal PstI site found at 7600
    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 4524
    Illegal SapI site found at 5814
    Illegal SapI.rc site found at 7011


Design Notes

The effectiveness, and GC content.


Source

The CRISPRi fragment is derived from PZ-ProP-Ect-i2 plasmid, then be ligated after EctOperon on the PZ-ProP-Ect-i2 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.