Composite

Part:BBa_K3726067

Designed by: Santiago Barragán Ariza   Group: iGEM21_MADRID_UCM   (2021-10-19)
Revision as of 01:33, 20 October 2021 by Sbarraga (Talk | contribs)

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Lv.1 HspA - 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_K3726061” L0_HspA which codifies the major heat shock protein capable of enhancing cellular fitness in the presence of environmental stresses such as solvent presence.

Within our project we have used HspA to reduce ROS stress generated from n-butanol. ("Using Transcriptomics To Improve Butanol Tolerance of Synechocystis sp. Strain PCC 6803 | Applied and Environmental Microbiology", Applied and Environmental Microbiology, 2021.)

The part in its downstream region has a homology region “BBa_K3726108” 5CON1(H)_NS3(mod)-up (PCC 11801) for homologous recombination within the genome of PCC 11801. In the upstream region, the part has the connector 3'Con1 “BBa_K2560070” that will allow the assembly of a Lv2 construct following the marburg collection standard.

Likewise, a chloramphenicol resistance cassette is included downstream the transcriptional unit region.

Expression is controlled by the strong constitutive promoter for cyanobacteria, PS-PR “BBa_K3726084” and the strong synthetic RBS, RBS_PtaBs “BBa_K3726092”

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 798
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 798
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 2654
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 798
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 798
    Illegal NgoMIV site found at 724
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None