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DarR

Part:BBa_K1045017:Design

Designed by: iGEM Team Göttingen 2013   Group: iGEM13_Goettingen   (2013-09-20)
Revision as of 14:24, 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,BBa_1045011 and BBa_1045012. The hybridization oligos were ordered from Sigma-Aldrich. Sequence information derived from the parts registry and Zhang et al., 2013. In addition, parts that were obtained either by amplification of plasmid DNA from the distribution kit 2013(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