Difference between revisions of "Part:BBa K4765018"
(→‎Sequencing map)
Line 17: Line 17:
 
| <html><img style="width:640px" src="https://static.igem.wiki/teams/4765/wiki/zsl/sequencing-map-of-fen1.jpg" alt="contributed by Fudan iGEM 2023"></html>
 
| <html><img style="width:640px" src="https://static.igem.wiki/teams/4765/wiki/zsl/sequencing-map-of-fen1.jpg" alt="contributed by Fudan iGEM 2023"></html>
 
|-
 
|-
−
| '''Figure 1. Sequencing map of FEN1''' Sequencing starts from the T7 terminator, with the primer 5-GCTAGTTATTGCTCAGCGG-3.
+
| '''Figure 1. Sequencing map of FEN1'''  
 +
Sequencing starts from the T7 terminator, with the primer 5-GCTAGTTATTGCTCAGCGG-3.
 
|}
 
|}
  

Revision as of 07:39, 10 October 2023


FEN1

contributed by Fudan iGEM 2023

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

contributed by Fudan iGEM 2023
Figure 1. Sequencing map of FEN1

Sequencing starts from the T7 terminator, with the primer 5-GCTAGTTATTGCTCAGCGG-3.

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 556
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1134
    Illegal AgeI site found at 240
  • 1000
    COMPATIBLE WITH RFC[1000]


Reference

  1. ↑ 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