Composite

Part:BBa_K1955010:Design

Designed by: Yu-chang Ku, Pang-ling Huang   Group: iGEM16_CGU_Taiwan   (2016-10-19)


pSB1A2-LacI inducible GFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 870


Design Notes

Use the standard parts provided by iGEM, we choose BBa_J04500 as a promoter to induce production of protein. BBa_J04500 is a LacI inducible promoter with RBS, so it can be induced by IPTG to open its promoter and start transcription. Double restriction enzyme cutting(EcoRI and XbaI) of pSB1A2-GFP to produce a sticky end in the anterior, then also double restriction enzyme cutting(EcoRI and SpeI) of BBa_J04500 to produce other sticky end which can be ligated with EcoRI and XbaI sticky end. Moreover, it will form a scar when XbaI ligate with SpeI, so we can simply compose the pSB1A2-J04500-GFP.


Source

LacI inducible promoter came from BBa_J04500; GFP protein sequence came from BBa_E0040.

References