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Coding

Part:BBa_K1045001:Design

Designed by: iGEM Team Göttingen 2013   Group: iGEM13_Goettingen   (2013-06-24)

DarR ORF with inversed Pre- and Suffix


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 581
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 594
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 31
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

In order to express DarR in Escherichia coli, the GTG-start codon was mutated to an ATG. Also, the first few codons were changed according to codon usage in E. coli via mutagenesis PCR (with the forward primer).

Source

This part was amplified from the genomic DNA of Mycobacterium smegmatis purchased from DSMZ ([http://www.dsmz.de/catalogues/details/culture/DSM-43756.html?tx_dsmzresources_pi5%5BreturnPid%5D=304 DSM No. 43756]).

References

Lei Zhang, Weihui Li, and Zheng-Guo He (2013) “DarR, a TetR-like Transcriptional Factor, Is a Cyclic Di-AMP-responsive Repressor in Mycobacterium smegmatis”, THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 288, NO. 5, pp. 3085–3096