Part:BBa_K4951070:Design
pTwist Kan High Copy mScarlet3 - Insert (Trigger)
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 132
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 132
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 132
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 132
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 132
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 23
Design Notes
The insert was ordered in a pTwist backbone from Twist Biosciences. The high copy origin of replication is intended to maximize the presence of the trigger gene in the cell for switch testing purposes. The switches we intended to test in this plasmid was design to target the KanR gene. mScarlet3 was chosen to serve as a reporter because of its exceptionally high fluorescent signal.
The mScarlet3 gene is driven by a T7 promoter and a strong RBS, retrieved from Dr. Nils Averesch. The insert has contains 11 random basepairs between the promoter and the RBS and 7 basepairs of spacing between the RBS and the open reading frame. We received advice and guidance from Dr. Alex Engel and Dr. Nils Averesch regarding the design of this plasmid.
Source
This plasmid insert was ordered from Twist Biosciences into a pTwist backbone.