DNA

Part:BBa_K4683000

Designed by: Natalie Tan, Manasvi Gupta, Vandana Vijay Kumar, Kate Sharer   Group: iGEM23_Lambert-GA   (2023-10-10)


Lettuce Complement

This part is the complement sequence of the fluorescent DNA aptamer Lettuce BBa_K4245133, developed by the Weill-Cornell Medical College. Lettuce is a 99 nucleotide long DNA aptamer that induces fluorescence of a GFP fluorophore mimic (Z)-4-(3,5-difluoro-4-hydroxybenzylidene)-2-methyl-1-(2,2,2-trifluoroethyl)-1H-imidazol-5(4 H)-one (DFHBI-1T) (VarnBuhler et al., 2022). This sequence can be combined with Nb.BbvCI nicking enzyme cut sites BBa_M31961 and complementary arms to become a padlock probe for exponential Rolling Circle Amplification (eRCA) and as a reporter. Due to the eRCA process creating short sequences of DNA, with this part as the middle sequence, Lettuce aptamers can be produced and quantified (Li et al., 2017).

References
Li, X.-Y., Du, Y.-C., Zhang, Y.-P., & Kong, D.-M. (2017). Dual functional phi29 DNA polymerase-triggered exponential rolling circle amplification for sequence-specific detection of target DNA embedded in long-stranded genomic DNA. Scientific Reports, 7(1). https://doi.org/10.1038/s41598-017-06594-1

VarnBuhler, B. S., Moon, J., Dey, S. K., Wu, J., & Jaffrey, S. R. (2022). Detection of SARS-CoV-2 RNA Using a DNA Aptamer Mimic of Green Fluorescent Protein. ACS chemical biology, 17(4), 840–853. https://doi.org/10.1021/acschembio.1c00893

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None