Composite

Part:BBa_K3888011

Designed by: Linhan Lei   Group: iGEM21_SJTang   (2021-10-01)
Revision as of 12:56, 20 October 2021 by RyanLei (Talk | contribs)


[Ni-Fe] Hydrogenase Core gene HoxG1 and HoxK1

This composite part is used for heterologous expression of oxygen-tolerant Hydrogenovibrio marinus [NiFe]-hydrogenase in Escherichia coli.
Hydrogenase is extremely sensitive to oxygen. By expressing oxygen-tolerant hydrogenase in E.coli, the oxygen tolerance will increase thus increases the amount of hydrogen produced by hydrogenase in E.coli. The strain of E.coli we used in the experiments is E.coli BL21. The part basically consists of two main gene: HoxG1 and HoxG1, which encode the large subunit and small subunit of oxygen-tolerant Hydrogenovibrio marinus [NiFe]-hydrogenase respectively. Other key features also include a natural (wild) type of RBS, B0032m, and a pTac promoter which is controlled by LacI(induced by IPTG) and lac operator .

PET28-pTac-lacO-hydrogenase .png

Figure 1: The Oxygen-tolerant Hydrogenase introduction plasmid constructed, which has a length of 8261bp.

Sequence and Features This composite part is used to express oxygen-tolerant hydrogenase, the lengths and positions of some important components are showed below. HoxG1 encodes the large subunit of hydrogenase and has a length of 1833bp, which is located from 5094bp to 6926bp on the plasmid diagram showed above. HoxK1 encodes the small subunit of hydrogenase and has a length of 966bp, which is located from 7137bp to 8102bp on the plasmid diagram showed above. RBS:B0032m has a length of 20bp, located from 5068bp to 5087bp. LacI has a length of 1083bp, located from 3515bp to 4597bp. ptas has a length of 29bp, located from 5006bp to 5034bp.


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 363
    Illegal XbaI site found at 465
    Illegal SpeI site found at 531
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 531
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 678
    Illegal BamHI site found at 1593
    Illegal BamHI site found at 2932
    Illegal XhoI site found at 1915
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 363
    Illegal XbaI site found at 465
    Illegal SpeI site found at 531
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 363
    Illegal XbaI site found at 465
    Illegal SpeI site found at 531
    Illegal AgeI site found at 1809
  • 1000
    COMPATIBLE WITH RFC[1000]


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