Difference between revisions of "Part:BBa K3192027:Design"

 
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===Design Notes===
 
===Design Notes===
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The 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.
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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.
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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.
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ggtctcNNNNNccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctcNNNNNggtctc
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Revision as of 21:29, 19 October 2019


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 87


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.

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.

ggtctcNNNNNccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctcNNNNNggtctc



Source

Escherichia coli

References