Reporter

Part:BBa_K3147003:Design

Designed by: Thomas Bessede   Group: iGEM19_Montpellier   (2019-10-13)

mRFP1 fused to a TEV-cleavable ssrA tag


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 691
  • 1000
    COMPATIBLE WITH RFC[1000]


Design note

The TEV cutting site used is the conventional site composed of 6 amino acids: ENLYFQS.

We have observed that this construction is weakly expressed. We advise future users to add a RiboJ insulator and a BCD2 element to improve its expression.

A double terminator controls the expression of this parts Part:BBa_B0015.

We placed the ssrA in C-ter because it is well known to be working on this side of the protein. [5]

This sequence was synthesized by IDT DNA for iGEM Headquarters.

Reference

[5] Sunohara, T., Abo, T., Inada, T., & Aiba, H. (2002). The C-terminal amino acid sequence of nascent peptide is a major determinant of SsrA tagging at all three stop codons. RNA (New York, N.Y.), 8(11), 1416–1427. doi:10.1017/s1355838202020198