Part:BBa_K4006012
MT-IEE5-arsC-IEE2-fTNa
Background
This composite part includes coding sequenceS for the proteinS metallothionein, arsenate reductase, and the protein ferritin, all codon optimized for use in the Chlamydomonas reinhardtii chloroplast. The intention was for them to work synergistically to improve the uptake of arsenic out of contaminated water.
Cloning into E. coli and Verification
The constructs were designed by our team and produced by IDT. They were transformed into 10-Beta competent E. coli cells from NEB. The “Full” sequence, MT-IEE2-ArsC-IEE5-FtnA, was designed by our team and produced in two parts by IDT. However, our team did not have any success cloning this construct via golden braid. This may be due to the presence of IEE2, which is also present in the pASapI backbone. We were unable to confirm cloning via restriction digest and often did not get significant results from the transformation into 10-beta competent E. coli cells.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 606
Illegal BamHI site found at 40 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 224
Illegal SapI site found at 1709
Illegal SapI.rc site found at 31
Illegal SapI.rc site found at 2219
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