Part:BBa_K4387986
Monovalent nanobody expression with the HlyA secretion signal (VHH#2B)
The hemolysin A secretion machinery is a one-step secretion system (T1SS), originally isolated from uropathogenic E. coli strains. It comprises three main peptides, the inner membrane proteins HlyB and HlyD, and the outer membrane protein TolC. Together, these three proteins build a continuous channel through which originally the HlyA toxin is secreted in a one-step manner. Scientists have identified the secretion signal and were able to secrete various proteins of different sizes with this secretion machinery. [1]
This composite part contains an anti-TNFa nanobody with a myc-tag and the HlyA-tag required for the secretion with the hemolysin A secretion system BBa_K4387979. The expression of the monovalent nanobody is under the control of the inducible pBAD system, followed by the T7 RBS. The myc-tag was required for the characterization of the secretion and functionality of the nanobody. The nanobody inserted in this part is the monovalent candidate VHH#2B BBa_K4387996. [2]
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1205
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1650
Illegal BglII site found at 2229
Illegal BamHI site found at 1144 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 979
- 1000COMPATIBLE WITH RFC[1000]
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