Circularly permutated haloalkane transferase fusion molecules

A modular polypeptide comprising or consisting essentially of a first partial effector sequence comprising or consisting essentially of

• an N-terminal first effector sequence part comprising SEQ ID NO:
• 002 or by a sequence at least (≥) 90% identical to SEQ ID NO:
• 002,
• a C-terminal first effector sequence part comprising SEQ ID NO:
• 003 or by a sequence at least (≥) 90% identical to SEQ ID NO:
• 003,
• an internal linker consisting of 10 to 35 amino acids, wherein the internal linker connects the C-terminus of the N-terminal first effector sequence part to the N-terminus of the C-terminal first effector sequence part;
• connected to
• a sensor module sequence, which is connected to
• a second partial effector sequence comprising or consisting essentially of a sequence selected from SEQ ID NO:
• 006 (PEP1), SEQ ID NO:
• 007 (PEP2), and a sequence at least (≥) 75% identical to SEQ ID NO:
• 007 (PEP2),
• wherein said sequence at least (2) 75% identical to SEQ ID NO:
• 007 (PEP2) has at least one mutation at position A151, R146, E147, T148, or T154 with respect to SEQ ID NO:
• 007 (PEP2),
• wherein
• the first and second partial effector sequences together constitute a circularly permuted haloalkane dehalogenase, and are capable, when brought into close proximity of each other, to effect covalent attachment of a halogen alkane moiety, and wherein
• the sensor module sequence is selected from
• a) a single sensor polypeptide capable of undergoing conformational change from a first confirmation to a second confirmation depending on the presence or concentration of an analyte compound, wherein in the first conformation, the first and second partial effector sequences are in close proximity, and in the second conformation, the first and second partial effector sequences are not in close proximity, wherein the first partial effector sequence is attached to the C-terminus of the sensor module sequence and the second partial effector sequence is attached to the N-terminus of the sensor module sequence,
• and
• b) a sensor polypeptide pair comprising a first sensor polypeptide and a second sensor polypeptide, wherein the first sensor polypeptide is covalently attached through a peptide bond to the first partial effector sequence and the second sensor polypeptide is covalently attached to the second partial effector sequence,
• the first sensor polypeptide and the second sensor polypeptide are capable of specific molecular interaction,
• and the first and second sensor polypeptides are part of separate polypeptide chains.