Part:BBa_K4103015:Design
Txn_ccdB
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 20
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 915
Illegal BsaI.rc site found at 14
Design Notes
we considered not including promoter and terminator, but since those change between organisms its wise to do so - we want to create sequences that are optimized for a specific use yet not too specific that theyre not usable across a wide enough scale
Q:I see that it uses BsaI to cut out the dropout cassette, and has BbsI cut sites for assembling into a vector. I'm curious how this works to insert the dropout cassette into the TU slot of an assembling pHT43-style vector being built with our parts--don't their level 1 digest reactions require BsaI? Apologies if I'm missing something obvious; running on not much sleep currently A:You are expecting that BsaI in a MoClo assembly is 100% efficient. It is not! Further you can drop the temp to 16oC for 10 minutes at the end to favour ligation over cutting. You will get assembly with the Dropout. It works!