Composite

Part:BBa_K1919101

Designed by: Zekun Lv   Group: iGEM16_SCU-China   (2016-10-14)


A temperature promoter cotaining clts repressor

This is based on part(BBa_K608351).Composite promoter (BBa_K608351) include the mutant type CIλrepressor so that it can express above 37℃.Compared with the part(BBa_K608351),it is more sensitive at 37℃.According to the essay, we choose five important sites A44G, G92A, C188T, G378A, A629T, which may enhance more than tenth times expression at 37℃，compared with wild type(BBa_K608351).

Tp1.png

Table1:Galactosidase acitivty unit after induction and compared with cIts857[1]


Tp2.png

Table2:Nucleotide changes and the deduced amino acid changes in five thermosensitive cI repressor proteins[2]

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NotI site found at 781
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


As for the measurement of this part, we will choice eGFP and FCM to test the expression at 37℃.

Reference

[1] [2] Using DNA shuffling method generate thermosensitive CI repressor protein ofλphage. Shen Guo-miao.June, 2003. Dissertation Submitted to Zhejiang University For Master’s Degree.

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