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Part:BBa_K4765018
FEN1
Introduction
FEN1 plays a pivotal role in DNA maintenance, addressing 5' overhanging "flaps" formed when DNA strands can't bind correctly. It also manages the 5' ends of Okazaki fragments in lagging strand DNA synthesis. This protein interacts directly with AP endonuclease 1 during long-patch base excision repair, enabling seamless substrate transfer between enzymes. Belonging to the XPG/RAD2 endonuclease family, it's one of ten essential proteins for cell-free DNA replication[1]. However, certain DNA structures can hinder its flap processing at trinucleotide repeats, concealing the crucial 5' end. This obstruction can compromise its protective function, potentially causing site-specific trinucleotide expansions linked to genetic disorders.
Usage and Biology
We heterologously expressed codon-optimized FEN1 in E. coli, endowing it with anti-UV capability.
Characterization
Sequencing map
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Figure 1. Sequencing map of FEN1
Sequencing starts from the T7 terminator, with the primer 5-GCTAGTTATTGCTCAGCGG-3. |
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 556
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1134
Illegal AgeI site found at 240 - 1000COMPATIBLE WITH RFC[1000]
Reference
- ↑ Hiraoka, L. R., Harrington, J. J., Gerhard, D. S., Lieber, M. R., & Hsieh, C. L. (1995). Sequence of human FEN-1, a structure-specific endonuclease, and chromosomal localization of the gene (FEN1) in mouse and human. Genomics, 25(1), 220–225. https://doi.org/10.1016/0888-7543(95)80129-a
protein |