Composite

Part:BBa_K3726071:Design

Designed by: Santiago Barragán Ariza   Group: iGEM21_MADRID_UCM   (2021-10-19)
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Lv.1 AcrbV2 - B


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1023
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 752
    Illegal PstI site found at 1023
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1023
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1023
    Illegal NgoMIV site found at 2159
    Illegal NgoMIV site found at 3675
    Illegal AgeI site found at 109
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The part is a MoClo Lv.1 part, assembled in the acceptor Lv1 entry vector “BBa_K3228069” P1_pANs_SpecR, which is a self replicating shuttle vector. This part has been assembled following the marburg collection standard, then the transcriptional unit is flanked by two connectors that will allow the assembly of a Lv2 construct. In its upstream region the connector is 5'Con2 “BBa_K2560066”, and in its downstream region 3'Con2 “BBa_K2560071”.

This design will allow us to test which “tolerance and secretion ” variant has better efficiency by its own account as a replicative vector, and then assemble them as an integrative Lv2 Moclo construct. To know more about this, visit our webpage: https://2021.igem.org/Team:MADRID_UCM/Design

Source

This construct has been made by golden gate reaction.

References

"Enhancing Tolerance to Short-Chain Alcohols by Engineering the Escherichia coli AcrB Efflux Pump to Secrete the Non-native Substrate n-Butanol", ACS Publications, 2021. [Online]. Available: https://pubs.acs.org/doi/10.1021/sb400065q.