Part:BBa_K2751008:Design
YPet skeleton
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 40
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 61
Illegal XhoI site found at 790 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 713
Design Notes
Use: The part can be put under a promoter to simply express YPet with his tag. Yet it can also be modified to produce YPet-fused proteins with cutting sites in prefix and suffix. If one wishes to simply acquire the sequence of YPet, cutting sites in the prefix and the suffix can also be applied.
Design notes: This part is not just a fluorescent protein. It contains ingeniously design prefix and suffix that make it easy-to-use and highly modifiable. The prefix contains a ribosome binding site(RBS), which means that once the part is put under a promoter and the promoter is induced, YPet would be expressed. The prefix also has many frequently-used cutting sites-NheI, KpnI, BamHI. This means that sequences that one wishes to express as a fusion protein with YPet can be easily inserted to the prefix and that this single and simple design of prefix can accommodate a wide range of sequences. In the suffix, polyhistidine tag is linked to the 3’ end of YPet, meaning that the protein expressed with this part automatically contains his tag and can be purified with Ni2+ chromatography. The suffix contains a SalI and an XholI cutting site as well, enabling the suffix to be inserted with other sequences. Lastly, using cutting sites in both the prefix and the suffix, one can cut YPet out of the sequence and put to other uses.
Source
We have acquired the sequence of YPet from Dr. Yaw-Kuen Li of NCTU-Taiwan. The prefix and the suffix are added by PCR.