Composite

Part:BBa_K4228999

Designed by: Keyao Zhao   Group: iGEM22_Tongji_China   (2022-09-29)
Revision as of 16:23, 11 October 2022 by Fork (Talk | contribs)

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P43-Bromelain-T7

Thinking of experimental requirements,we chose which is cheep and common to finish our experiment.

In order to complete the heterogenous expression of mutant bromelain in Bacillus subtilis, we cloned the sequence as BamHI-XhoI inserts in the pBER-2 expression vector as well. pBER-2 is a P43 promoter based shuttle vector which can propagate in Escherichia coli and Bacillus subtilis.The desired polypeptide can be expressed as a fusion protein with His6 tag at the C-terminus and Alkaliophilic Bacillus alkaline protease signal peptide and propeptide at the N-terminus to induce protein secretion.


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 312
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 312
    Illegal NheI site found at 1120
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 312
    Illegal BamHI site found at 1
    Illegal XhoI site found at 646
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 312
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 312
  • 1000
    COMPATIBLE WITH RFC[1000]


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