Coding

Part:BBa_K2455002:Design

Designed by: Jon Fugl   Group: iGEM17_UCopenhagen   (2017-10-25)
Revision as of 00:47, 2 November 2017 by Jon UCPH (Talk | contribs) (References)


CPP-SYFP2


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]


Design Notes

The first step in the insertion was amplification of SYFP2 using primers matching the USER Cassette of the [[Part:BBa_K2455003|Cell-Penetrating USER Cassette]. To do this we used the following primers:

Forward primer: CGTGCGAUCA-ATGGTTAGCAAGGGCGAAG

Reverse primer: CACGCGAUGA-TTTATACAGCTCATCCATACCCAGGG

Subsequently the PCR product could be gel-extracted and ligated into the Cell-Penetrating USER Cassette biobrick.

Finally for the creation of a biobrick version the following set of primers were used:

Forward primer: ATCCGGAATTCGCGGCCGCTTCTAG-ATGCGTCGCCGTCGCCGTCGCCGTCGCCGTGAATGCGTGCGATC

Reverse primer: CCAATGCATTGGTTCTGCAGCGGCCGCTACTAGTATTATTA-ATGGTGATGGTGATGATGTAAG

Source

The biobrick is an improved version of the Yellow Fluorescent Protein 2 (SYFP2).

References

Chang, M. et al. (2005). Cellular Internalization of Fluorescent Proteins via Arginine-rich Intracellular Delivery Peptide in Plant Cells. PCP Plant and Cell Physiology, 46(3), pp. 482-8.

Eudes & Chugh (2008). Cell-penetrating peptides. Plant Signal Behav., 3(8), pp. 549–550.