Composite

Part:BBa_K3726069

Designed by: Santiago Barragán Ariza   Group: iGEM21_MADRID_UCM   (2021-10-19)


Lv.1 SigB - B

This is a composite part that corresponds with a level 1 MoClo part in accordance with the marburg collection standard. It is a transcriptional unit for the expression of the part “BBa_K3726063” L0_SigB which codifies a sigma factor that when it is over-expressed increases tolerance to temperature, and reduces ROS stress. Within our project we have used SigB to reduce ROS stress generated from n-butanol.

The part in its upstream region, has the connector 5'Con3 “K2560067” that will allow the assembly of a Lv2 construct following the marburg collection standard. In its downstream region has a homology region “BBa_K3726109” 3CON5(H)_NS3(mod)-down (PCC 11801) for homologous recombination within the genome of PCC 11801.

Expression is controlled by the medium-strong synthetic constitutive promoter J23119 “BBa_K2560031”and the strong synthetic RBS, RBS_PtaBs “BBa_K3726092”

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 850
    Illegal PstI site found at 1752
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 850
    Illegal NheI site found at 431
    Illegal NheI site found at 454
    Illegal NheI site found at 1477
    Illegal PstI site found at 1752
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 850
    Illegal XhoI site found at 886
    Illegal XhoI site found at 1123
    Illegal XhoI site found at 1905
    Illegal XhoI site found at 2025
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 850
    Illegal PstI site found at 1752
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 850
    Illegal PstI site found at 1752
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 610


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