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DarR

Part:BBa_K1045017:Design

Designed by: iGEM Team Göttingen 2013   Group: iGEM13_Goettingen   (2013-09-20)
Revision as of 14:21, 19 October 2013 by Kati (Talk | contribs) (→‎Source)

DarR reporter system


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 806
    Illegal NheI site found at 829
    Illegal NheI site found at 909
    Illegal NheI site found at 932
    Illegal NotI site found at 718
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 731
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 860
    Illegal AgeI site found at 168
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1630


Design Notes

DarR fused to the terminator was cut out from BBa_K1045016 and ligated in a pre-fixing composition into BBa_K1045015.

Source

This composite part was constructed using hybridization oligos for BBa_1045010 and BBa_1045011. The hybridization oligos were ordered from Sigma-Aldrich. In addition, parts that were obtained either by amplification of plasmid DNA (inverse terminator BBa_1045009) or M. smegmatis chromosomal DNA (DarR ORF, BBa_1045001 were employed. The chromosomal DNA of Mycobacterium smegmatis was purchased from DSMZ ([http://www.dsmz.de/catalogues/details/culture/DSM-43756.html?tx_dsmzresources_pi5%5BreturnPid%5D=304 DSM No. 43756]). Part BBa_E0240 derived from the plasmid in the distribution kit 2013.

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