Difference between revisions of "Part:BBa K1045017:Design"

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===Design Notes===
 
===Design Notes===
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The DarR-encoding gene was fused to the terminator and cut out from [[Part:BBa_K1045016|BBa_K1045016]] and ligated in a pre-fixing composition into [[Part:BBa_K1045015|BBa_K1045015]].
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The insert [[Part:BBa_K1045015|BBa_K1045015]] was cut out from its plasmid and ligated in a post-fixing composition into the vector [[Part:BBa_K1045016|BBa_K1045016]].
  
 
===Source===
 
===Source===

Latest revision as of 20:06, 28 October 2013

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

The insert BBa_K1045015 was cut out from its plasmid and ligated in a post-fixing composition into the vector BBa_K1045016.

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 Mycobacterium smegmatis chromosomal DNA (DarR ORF, BBa_1045001) were employed. The chromosomal DNA of M. smegmatis was purchased from DSMZ, Braunschweig, Germany ([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