Part:BBa_K2779912
L-arabinose-induced eGFP with 6xHis tag
Team UAlberta’s cloning efforts to clone our protoporphyrin IX biosynthesis pathway relied heavily on the use of visible and fluorescent markers to assemble our desired constructs. In order to characterize our enzymes, we wanted a system that would enable us to easily clone, express and purify each of our enzymes for further experimentation.
We wanted an araBAD-based expression system because:
It is inducible by L-arabinose and is tightly regulated by araC; It consists of a strong promoter for increased expression; It is compatible with our preferred workhorse strain, DH10B, allowing us to perform our cloning and expression work in the same strain; and It is more cost-effective, since L-arabinose is more economical than IPTG. When we initially looked through the repository, we found that while <a href="https://parts.igem.org/Part:BBa_K1602055"> BBa_K1602055</a>, which had GFP under the control of araC/pBAD, was the closest to our needs, there was no easy way for us to replace their inserted GFP with our desired genes, and there was no way for us to collect the expressed enzymes. Thus, we decided to improve BBa_K1602055.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1476
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1346
Illegal BamHI site found at 1532
Illegal XhoI site found at 1549 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1181
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1398
Illegal SapI site found at 1163
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