Difference between revisions of "Part:BBa K2429183:Design"
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===Design Notes=== | ===Design Notes=== | ||
| − | + | Due to the gene's large size, dCas13a had many restriction sites that would not make it BioBrick Compatible. To account for this, our team decided to order g-blocks of dCas13a with slightly altered sequences to rid if prohibited restriction sites rather than apply multiple point mutations to the existing construct. After the g-blocks arrived, a teammate applied Gibson Assembly to put them together in the pSB1C3 backbone. | |
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===Source=== | ===Source=== | ||
Latest revision as of 22:29, 26 October 2017
L. shahii Cas13a BioBrick
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 572
Illegal BglII site found at 1196
Illegal BglII site found at 1592
Illegal BglII site found at 1883
Illegal BglII site found at 1973
Illegal BglII site found at 3134
Illegal BglII site found at 3221
Illegal BamHI site found at 4216 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 4173
Illegal NgoMIV site found at 4192 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 558
Illegal SapI.rc site found at 664
Illegal SapI.rc site found at 1534
Illegal SapI.rc site found at 2368
Design Notes
Due to the gene's large size, dCas13a had many restriction sites that would not make it BioBrick Compatible. To account for this, our team decided to order g-blocks of dCas13a with slightly altered sequences to rid if prohibited restriction sites rather than apply multiple point mutations to the existing construct. After the g-blocks arrived, a teammate applied Gibson Assembly to put them together in the pSB1C3 backbone.
Source
Sequence for L. shahii Cas13a was found from Addgene(https://www.addgene.org/79150/), which is also where our team received the original plasmid from.