Coding

Part:BBa_K1349004:Design

Designed by: Alexia Satouf, Clara Bouyx, Aimeric Agaoua, Lambert Antoni, Vincent Castel   Group: iGEM14_Aix-Marseille   (2014-10-06)
Revision as of 07:40, 9 October 2014 by LISM JS (Talk | contribs) (→‎Source)

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mesh1


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 512
    Illegal BamHI site found at 78
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part was designed to allow rapid degradation of the alarmone ppGpp, responsible for the stingent response in E. coli. In the context of our project ppGpp degradation is used to start initiation of cell division. This part is complementary to part BBa_K1349001.

The part was obtained by PCR from pBAD-Mesh1 plasmid kindly provided by Ben Field and SLIC assembly. The construction removed the PstI restriction sites in the gene by the silent mutations of a CAG codon to a CCG codon. The construction also removed the SpeI restriction sites in the gene by the silent mutations of a ACT codon to a TCT codon.The construction contains the entire coding sequence of the Drosophila gene.

Expression is expected to cause destruction of ppGpp. Expression in a mutant overproducing ppGpp is expected to complement the mutant.

Source

The construction contains the entire coding sequence of the Drosophila gene. The part was obtained by PCR from pBAD-Mesh1 plasmid kindly provided by Ben Field.

References