Composite

Part:BBa_K3893012:Design

Designed by: Tannya Sandoval   Group: iGEM21_Ecuador   (2021-10-21)


Up part of cassette for dsRNA constitutive production 3


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 2525
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 749
    Illegal NheI site found at 1665
    Illegal NheI site found at 2470
    Illegal NheI site found at 3279
    Illegal SpeI site found at 2525
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2519
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 2525
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 2525
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 665
    Illegal BsaI.rc site found at 877
    Illegal BsaI.rc site found at 1581
    Illegal BsaI.rc site found at 2386
    Illegal BsaI.rc site found at 3195


Design Notes

In order to use, test and produce the dsRNA we design before, we created a plasmid backbone with an insert that is able to receive different sequences of dsRNA with a BglII and KpnI digestion.

With this in mind, we face some shortcomings: first, it is a short fragment, with many sequence repeats, which makes it difficult for synthesis. Second, we want this insert to behave as a Biobrick compatible once it has the desired dsRNA sequence cloned inside.

It isn't compatible with RFC10 Assembly. Because the SpeI and BglII restriction sites are needed in the middle of the sequence for the assembly system.


Source

Set of biobricks