Part:BBa_K4960023

Pdp1_NTD-3*GGSGG-Cre-GSSG-HiBiT
This part is responsible for payloads loading Cre.

Profile

Name: Pdp1NTD-3*GGSGG-Cre-GSSG-HiBiT
Base Pairs: 1317 bp
Origin: Photorhabdus, synthetic

Usage and Biology

This part includes the packaging domain of PVC and the gene encoding protein Cre and polypeptide HiBiT.[1]Cre is a 38kDa protein composed of 343 amino acids, with 4 subunits and 2 domains: a larger carboxyl (C-terminal) domain and a smaller amino (n-terminal) domain. It is a site-specific DNA recombination enzyme that can specifically recognize loxp sites and mediate sequence deletion or recombination between loxp sites. The sequence of loxp is as follows:
5' - ATAACTTCGTATA - GCATACAT - TATACGAAGTTAT - 3'
3' - TATTGAAGCATAT - CGTATGTA - ATATGCTTCAATA - 5'
HiBiT is a polypeptide tag, can be linked to any protein.
This part can achieve the purpose of mediating Cre protein injection into target cells and gene editing at specific loci in target cells.

Special Design

1.The linker which contains three or two repeating amino acid sequences links the packaging domain and another DNA fragment. After gene transcription and translation, in protein structures, linker sequences are usually flexible because the domains they connect to often have different conformation and motor requirements. Therefore, linker sequences often act as flexible connections, allowing the domain to move freely while maintaining structural stability.
2.The polypeptide tag, HiBiT, can be linked to any protein, enabling highly quantitative and extremely sensitive bioluminescence detection. This allows quantification of proteins using bioluminescence without the use of antibodies. With only 11 amino acids, HiBiT is easy to combine with gene-editing technology to precisely insert into the genome, allowing researchers to study proteins under physiologically relevant conditions.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 599
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 285
  
• 1000
  COMPATIBLE WITH RFC[1000]

References

[1] Joseph Kreitz, Mirco J. Friedrich, Akash Guru, Blake Lash, Makoto Saito, Rhiannon K. Macrae & Feng Zhang.Programmable protein delivery with a bacterial contractile injection system. Nature. Mar.9,357(2023)https://doi.org/10.1038/s41586-023-05870-7