Difference between revisions of "Part:BBa K1979003:Design"

(→‎Design Notes)
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===References===
 
===References===
 +
Karig et al. 2012;
 +
Lentini et al. 2013;
 +
Takahashi et al. 2013;

Revision as of 07:36, 28 August 2016


PBP5 coding device (promoter+PBP5+T7 terminator)


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 1178
    Illegal BamHI site found at 168
    Illegal XhoI site found at 1386
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 492
    Illegal AgeI site found at 1094
  • 1000
    COMPATIBLE WITH RFC[1000]


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 and terminator sequences are retrieved form GenBank, and the promoter sequence comes from the part BBa_J23101.


References

Karig et al. 2012;

Lentini et al. 2013; Takahashi et al. 2013;