Composite

Part:BBa_K3718012

Designed by: Chan Pui Ching   Group: iGEM21_Hong_Kong_UCCKE   (2021-10-04)
Revision as of 17:09, 16 October 2021 by Stephy0521 (Talk | contribs)


Device for knocking out arcA and iclR genes


Part I of toolkit for Detection with Proliferation Control

The first part of the toolkit contains two sgRNA (arcA and iclR) which will be knocked out to reduce cell growth.

To create the knockout strain for the toolkit, a plasmid with Cas9 gene and sgRNA that targets arcA and iclR is transformed into the wild-type E. coli. The knock out is done so that the cells have a lower growth rate than wild type (-38%). The repression is needed to allow a more acute cell proliferation and protein expression of the target gene the developer cloned into the second plasmid.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 4357
    Illegal suffix found in sequence at 4527
    Illegal EcoRI site found at 4599
    Illegal XbaI site found at 4614
    Illegal SpeI site found at 4770
    Illegal PstI site found at 4784
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4357
    Illegal EcoRI site found at 4599
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 4320
    Illegal NheI site found at 4343
    Illegal NheI site found at 4562
    Illegal NheI site found at 4585
    Illegal SpeI site found at 4528
    Illegal SpeI site found at 4770
    Illegal PstI site found at 4542
    Illegal PstI site found at 4784
    Illegal NotI site found at 4363
    Illegal NotI site found at 4535
    Illegal NotI site found at 4605
    Illegal NotI site found at 4777
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4357
    Illegal EcoRI site found at 4599
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 4357
    Illegal suffix found in sequence at 4528
    Illegal EcoRI site found at 4599
    Illegal XbaI site found at 4614
    Illegal SpeI site found at 4770
    Illegal PstI site found at 4784
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 4357
    Illegal EcoRI site found at 4599
    Illegal XbaI site found at 4372
    Illegal XbaI site found at 4614
    Illegal SpeI site found at 4528
    Illegal SpeI site found at 4770
    Illegal PstI site found at 4542
    Illegal PstI site found at 4784
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 3573


Reference

H. Waegeman, J. Beauprez, H. Moens, J. Maertens, M. De Mey, M. R. Foulquié-Moreno, J. J. Heijnen, D. Charlier, and W. Soetaert, “Effect of ICLR and ARCA knockouts on biomass formation and metabolic fluxes in escherichia coli K12 and its implications on understanding the metabolism of escherichia coli BL21 (DE3),” BMC Microbiology, vol. 11, no. 1, p. 70, 2011.

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