Composite

Part:BBa_K2382006:Design

Designed by: SHAO-CHI LO   Group: iGEM17_CSMU_NCHU_Taiwan   (2017-10-26)
Revision as of 22:59, 31 October 2017 by Ptrlintw (Talk | contribs) (→‎Design Notes)


T7 promoter + Thioredoxin-MSMEG_5998 fusion protein


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 456
    Illegal XhoI site found at 462
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 901
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

For the purpose of purification through nickel-resin column, we added a 18-bp sequence which can code 6 histidines. In addition, we chose T7 promoter which contains lac operator to express this protein because it can be induced by IPTG. For the terminator, we chose BBa_B0015 because it was commonly used in E. coli. The gene design are shown in Fig. 1.


Fig. 1


Source

References

1. Taylor, M.C., et al., Identification and characterization of two families of F420H2‐dependent reductases from Mycobacteria that catalyse aflatoxin degradation. Molecular microbiology, 2010. 78(3): p. 561-575.