Coding

Part:BBa_K2973020:Design

Designed by: Leandros Tsiotos, Vasiliki Kavvatha   Group: iGEM19_Thessaly   (2019-09-28)


TEM-optimized beta-lactamase / J23100


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 163
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 783


Design Notes

In order to attain maximum enzymatic activity, we implemented a strong protein expression promoter (BBa_J23100) and an optimized coding sequence for beta-lactamase. For this purpose we cloned the part in pSB1C3 and pSB1K3 high-copy plasmids and in E. coli DH5a cells. For protein expression, we transformed the plasmids into BL21 (DE3) and M15-T7 cells.

Source

https://parts.igem.org/wiki/index.php/Part:BBa_I757010

https://parts.igem.org/Part:BBa_B0034

https://parts.igem.org/wiki/index.php?title=Part:BBa_J23100

References