Difference between revisions of "Part:BBa K3726070:Design"
 
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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
 
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
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For last, regarding to P_pBAD (BBa_K3726076), the Sapl site should be removed in order to make this part RFC1000 compatible. however, this part is MoClo compatible with the marburg collection.
  
 
===Source===
 
===Source===

Latest revision as of 14:20, 20 October 2021


Lv.1 AcrbV2 - A


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 2033
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1634
    Illegal NheI site found at 1762
    Illegal PstI site found at 2033
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1573
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 2033
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 2033
    Illegal NgoMIV site found at 3169
    Illegal NgoMIV site found at 4685
    Illegal AgeI site found at 109
    Illegal AgeI site found at 1408
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1390


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 K2560066, and in its downstream region 3'Con2 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

For last, regarding to P_pBAD (BBa_K3726076), the Sapl site should be removed in order to make this part RFC1000 compatible. however, this part is MoClo compatible with the marburg collection.

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.