DNA
BsaI ->

Part:BBa_K1362424

Designed by: Constantin Ahlmann-Eltze, Charlotte Bunne, Magdalena Büscher, Jan Gleixner, Max Horn, Anna Huhn, Nils Klughammer, Jakob Kreft, Elisabeth Schäfer, Carolin Schmelas, Silvan Schmitz, Max Waldha   Group: iGEM14_Heidelberg   (2014-10-07)


BsaI restriction site for RFC[105] cloning

This is the reverse complement of a BsaI restriction site headed by an Adenine as a spacer-base to separate the recognition sequence from the outward-lying cutting sequence. It was used by us for scarless golden-gate cloning to fuse inteins to other proteins and thereby implement a variety of possible post-translational modifications.

Information added by Team IISER-Pune-India 2020

Team: IISER-Pune-India 2020

Author: Avadhoot Jadhav

Summary

We aimed to use this part for circularization of the cyclotide protein. For using this we found that it had 7 nucleotide sequences that made it incompatible for Gibson's assembly/Restriction free cloning. The 7 length sequence makes it out of frame with other components of the composite structure. So without disturbing the restriction site we designed a similar part that overcomes this problem: BBa_K3582021.

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1


[edit]
Categories
Parameters
None