Part:BBa_K2009363:Design
PGAL+GFP
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 70
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1119
Design Notes
The designer of Part:BBa_J63006 is very thoughtful for adding the KOZAK sequence to the downstream of the GAL1 promoter. Apparently, they meaned to improve the effeiciency of this part. However, considering the existence of the Biobrick prefix, the ATG inside the KOZAK sequence won't be in the same reading frame as the ATG of the downstream coding sequence. There were two teams managed to fix this problem, with the idea to eliminate the ATG inside the KOZAK sequence. This year, we applyed a different idea. We added one base pair between those two ATGs and made them belong to one common frame. Meanwhile, another ATG is made up between the former ones, belonging to the same frame.
Source
We construted this composite parts by ligating two former existing parts and doing a site mutation. The two parts we used is (1) GAL1 promoter Kozak sequence (Part:BBa_J63006) and (2) GFP (Part:BBa_E0040).
References
We construted BBa_K2009363 composite by ligating GAL1 promoter + Kozak sequence (Part:BBa_J63006) and (2) GFP (Part:BBa_E0040). However, the ATG inside the KOZAK sequence won't be in the same reading frame as the ATG of the downstream coding sequence, so a frame-shift mutation is inevitable. To solve this problem, we add a base pair after the Kozak sequence so that the ATG can be in the same reading frame and the GFP can be expressed properly. What’s more, this part become “ready to use”, which means the GFP sequence can be directly altered by other functional parts and the sequence will be expressed properly.