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Composite

Part:BBa_K1045016:Design

Designed by: iGEM Team Göttingen 2013   Group: iGEM13_Goettingen   (2013-09-20)

Terminator BBa_B0015 with inversed Pre- and Suffix- DarR ORF with inversed Pre- and Suffix


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    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 AgeI site found at 168
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The inverse DarR sequence was cut out from BBa_K1045001 and ligated in a post-fixing composition into the plasmid carrying BBa_K1045009.

Source

For this composite part, part BBa_K1045001 was used. Its sequence originated from PCR amplification from M. smegmatis chromosomal DNA. The genomic 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_K1045009 originated from PCR amplification of plasmid DNA derived from 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