Part:BBa_K1979003:Design
PBP5 coding device (promoter+PBP5+T7 terminator)
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 135 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1178
Illegal BamHI site found at 168
Illegal XhoI site found at 1386 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 492
Illegal AgeI site found at 1094 - 1000COMPATIBLE 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;