Part:BBa_K1251000
RiAFP
This is a composite part consisting of a T7 promoter containing a lac operator, a RBS, the coding sequence for the antifreeze protein, RiAFP, a 6x His tag, a secretion peptide, and a double terminator. This part controls the expression of RiAFP. The expressed protein is tagged with an amino acid motif that consists of six histidine (His) residues fused to the C-terminus of RiAFP. Histidine tags are widely used because they are small and rarely interfere with the function, activity, or structure of target proteins. The polyhistidine-tag is to be used to detect the secreted protein via anti-polyhistidine-tag antibodies or alternatively by in-gel staining (SDS-PAGE) with fluorescent probes bearing metal ions. The HlyA is a signal peptide found in the C-terminal signal sequence of alpha-hemolysin (HlyA). It is used to target RiAFP for secretion via the Type I secretion pathway of gram-negative bacteria. Fusion of the HlyA signal peptide to RiAFP results in transport of the protein from the cytoplasm to the extracellular medium in a single step
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1
Illegal PstI site found at 632
Illegal PstI site found at 859 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1
Illegal NheI site found at 185
Illegal PstI site found at 632
Illegal PstI site found at 859 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1
Illegal PstI site found at 632
Illegal PstI site found at 859 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1
Illegal PstI site found at 632
Illegal PstI site found at 859 - 1000COMPATIBLE WITH RFC[1000]
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