Part:BBa_K2117005:Design
Device encoding TEF1 and hrGFP for expression in Yarrowia lipolytica
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 2
Design Notes
The promoter was synthesized based on the native Y. lipolytica TEF1 promoter sequence, except for substitution of three bases to remove illegal restriction sites (PstI 166 G ->C, SpeI 201 T->A, ApoI 260 C->G). The sequence was subjoined with the RFC10 prefix and suffix.
The sequence of the hrGFP was obtained from Genbank and then codon optimized for Y. lipolytica and simultaneously removing illegal restriction sites. The hrGFP was made as a synthetic gene with the RFC10 prefix and suffix sequences in the ends. By standard 3A assembly the part was successfully assembled with the TEF1 promoter and assembled in the Y. lipolytica backbone vector, pSB1A8YL (BBa_K2117009). The hrGFP expression was visualized by Confocal Laser Fluorescent microscopy.