Generator

Part:BBa_K1979005:Design

Designed by: MA Xinyi   Group: iGEM16_SDSZ_China   (2016-09-17)


CBD-PBP5


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 135
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1322
    Illegal BamHI site found at 168
    Illegal XhoI site found at 1530
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 636
    Illegal AgeI site found at 1238
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 184


Design Notes

Histidine tag is added for affinity purification to harvest the proteins. g10-L is a strong RBS in E. coli but also well suited for foreign gene expression.The effect of the ribosome binding site (RBS) on in vitro translation has been investigated extensively (Karig et al. 2012; Lentini et al. 2013; Takahashi et al. 2013). Most important for an efficient reaction is a defined distance (4-9 bases) of the RBS to the following start codon (Karig et al. 2012; Lentini et al. 2013), but the strength of the RBS has a minor effect as long as it is not too weak (Karig et al. 2012; Takahashi et al. 2013).


Source

The PBP5 is from the E.coli's genes, CBD sequence is retrieved from GenBank, and the promoter is from part BBa_J23101.

References