Coding

Part:BBa_K2455003:Design

Designed by: Jon Fugl   Group: iGEM17_UCopenhagen   (2017-10-27)
Revision as of 23:25, 1 November 2017 by Jon UCPH (Talk | contribs) (→‎References)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)


Cell-Penetrating USER Cassette


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 biobrick was created by amplifying a modified version of the pET102 vector with the following set of primers, followed by blunt ligation:

Forward primer: CGTCGCCGTCGCCGT-GAATGCGTGCGATCGCG

Reverse primer: GCGACGGCGACGCAT-GCGTATGTATATCTCCTTCTTAAAG

For creation of a biobrick version the following set of primers was sub-sequently used:

Forward primer: ATCCGGAATTCGCGGCCGCTTCTAG-ATGCGTCGCCGTCGCCGTCGCCGTCGCCGTGAATGCGTGCGATC

Reverse primer: CCAATGCATTGGTTCTGCAGCGGCCGCTACTAGTATTATTA-ATGGTGATGGTGATGATGTAAG

Source

The nona-arginine CPP-tags is of synthetic origin, and is inspired by viral transduction domains.

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.

http://www.cbs.dtu.dk/services/AMUSER/

http://www.cbs.dtu.dk/services/AMUSER/instructions.php