Part:BBa_K3237004
His-mRFP1-Proinsulin with furin cut sites codon optimized for C. reinhardtii
This part is for the expression of insulin in microalgae, specifically Chlamydomonas reinhadtii . In its design there are the following synthetic components. The proinsulin part of the construct has been modified in that the original protease cut sites in proinsulin has been replaced with the Furin protease cut site as proposed by Tsinghua 2014. Furin is a also a human protein that will still allow for protein processing in an active form [1]. The second component is that this proinsulin is fused with red fluorescent protein (mRFP1) with a furin cut site and linker in between for removal. The third component is the addition of a his tag on the N-terminus of the mRFP1 to allow for purification from microalgae if necessary (although not proposed as a part of our project system). In summary, the part has a his tag on the N-terminus, which is attached to the mRFP1-proinsulin fusion. This fusion can be broken with the introduction of furin protease, and proinsulin will be processed to an active form simultaneously with the furin addition.
This part is codon optimized for use in C. reinhardtti but has a construct counterpart codon optimized for E.coli , BBa_K3237002.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 363
Illegal XhoI site found at 904 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
| None |