Terminator

Part:BBa_K3192026:Design

Designed by: Simonne Guenette   Group: iGEM19_Virginia   (2019-10-14)


Double terminator designed for golden gate assembly


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
    Illegal BsaI site found at 136


Design Notes

The recognition sequence ggtctc is used for recognition of the BsaI restriction enzyme used for Golden Gate assembly. When cloning multiple inserts into a vector using Golden Gate assembly, it is recommended to add a 5 base pair recognition sequence following the restriction site, before you insert, that is complementary to the adjacent insert of interest to increase the specificity of the reaction.

Figure depicting Golden Gate Assembly. Retrieved from https://www.neb.com/products/e1600-neb-golden-gate-assembly-mix#Product%20Information























For many golden gate assembly mastermixes the addition of a 5 base pair recognition sequence is recommended. This sequence should be complementary on each end with the adjacent overlapping sequence. Ensure that this sequence your team chooses to insert is not another restriction enzyme site. Below is an example of the sequence with the 5 unique sites inserted.

ggtctcNNNNNccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttataNNNNNggtctc


Source

Escherichia coli

References