Coding

Part:BBa_K2374002:Design

Designed by: Qian Zeng   Group: iGEM17_Tongji_China   (2017-10-20)
Revision as of 20:33, 30 October 2017 by Zjeng (Talk | contribs) (→‎Design Notes)


GAL80ts (temperature dependent)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 993
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 19
    Illegal BglII site found at 615
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 27
    Illegal BsaI site found at 73


Design Notes

We use PCR to clone the GAL80ts from the genomic D We connected TH promoter to GAL4( BBa_K2374004 [1] )and GAL80ts( BBa_K2374002 [2] )respectively, and cloned them into pUAST vector which had been removed the UAS sequence( BBa_K2374008 [3] ), then to microinject them into Drosophila 's eggs. Also we did microinjection with UAS-TH (BBa_K2374003 [4]). After hybridization screening, we got stable modified fruit fly strains. Finally, we did RT-PCR, qPCR and behavioral experiments to test our system. Here shows some results[http://2017.igem.org/Team:Tongji_China/Experiments].

pleP-GAL4


pleP-GAL80ts


pleP-GAL80ts



We cloned GAL80ts into pSB1C3 for submission. Here shows the 1% Agarose gel electrophoresis image.

标题

We did 2 mutagenesis on this sequence.
site direct mutagenesis:
1. EcoR I (184) GAATTC->GATTTC
2. Xba I (219) TCTAGA->TGTAGA

Source

Chromosome: XIII; NC_001145.3 (171594..172901) NOTE: Gal80ts is a mutation based on the gene above.


References