Difference between revisions of "Part:BBa J100419"

Revision as of 14:38, 5 August 2018

rClone mScarlet

This is the same sequence as the original rClone Red plasmid, but this plasmid has the mScarlet fluorescent protein inserted in place of the RFP. rClone Red: https://parts.igem.org/Part:BBa_J119384 mScarlet: https://parts.igem.org/Part:BBa_J100398

rClone mScarlet (see Eckdahl et al. 2017) allows users to clone and test new RBS elements and riboswitches without gel purification or other preparation of DNA. It is a destination vector for Golden Gate Assembly (GGA) using BsaI and ligase. A new RBS or riboswitch can be derived from synthetic oligos, PCR, or a plasmid clone. For proper assembly, the new insert must have the appropriate 4 nt sticky ends or be flanked by BsaI sites that produce the sticky ends. With reference to the top strand, the left site must be 5' CGAC 3' and the right site must be 5' GCGG 3'. BsaI always produces a 5' overhang sticky end. Successful GGA assembly replaces the reverse promoter and GFP with the new RBS or riboswitch. Transcription will be initiated in the direction of the mScarlet coding sequence. The level of expression of mScarlet will depend on the efficiency of the newly cloned RBS or riboswitch.

Sequence and Features