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Part:BBa_K2244007

Designed by: Chen Hong   Group: iGEM17_SSTi-SZGD   (2017-10-25)
Revision as of 17:16, 25 October 2017 by Qiucheng (Talk | contribs)

ColE promoter +mhei gene +mcherry gene +T1terminator


The device is a functional plasmid containing a MBC hydrolase gene (mheI) (BBa_K2244004), which encodes carbendazim hydrolase. In order to study expression of target gene in cytoplasm, mheI was fused to a reported gene mCherry (BBa_J06504), which functions as indicator of the uptake of mheI. This part is regulated under the control of ColE promoter, which is from E. coli ColE1 plasmid.


Biology

ColE promoter(BBa_K2244006) sequence is derived from the promoter region of colicin E gene located in the ColE1 plasmid of E.coli. ColE promoter contains a ‘SOS’ operator region that allows the binding of LexA protein to repress transcription.

MBC hydrolase gene (mheI)(BBa_K2244004) is derived from carbendazim-degrading strain Mycobacterium sp. SD-4 and is a genomic DNA gene sequence. mheI gene is also evolutionary conserved in many other bacteria strains, such as Nocardioides sp. SG-4G, and Rhodococcus erythropolis.

mCherry(BBa_K2244008) is a red fluorescent protein used as a reporter. It is based on a fluorescent protein that was originally isolated from Discosoma sp.mCherry sequence is codon optimized for E. coli expressionm and can be replaced by any gene sequence in light-regulated studies.

T1 terminator(BBa_B0010) is the most commonly used terminator.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1250
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1250
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 754
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1250
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1250
    Illegal AgeI site found at 413
  • 1000
    COMPATIBLE WITH RFC[1000]


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