Part:BBa_K4951072:Design
pTwist Amp Med Copy mStayGold + LacZ - Insert (Switch)
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 263
Illegal EcoRI site found at 955
Illegal XbaI site found at 236
Illegal PstI site found at 224 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 263
Illegal EcoRI site found at 955
Illegal PstI site found at 224 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 263
Illegal EcoRI site found at 955
Illegal BamHI site found at 242 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 263
Illegal EcoRI site found at 955
Illegal XbaI site found at 236
Illegal PstI site found at 224 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 263
Illegal EcoRI site found at 955
Illegal XbaI site found at 236
Illegal PstI site found at 224 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 608
Illegal SapI.rc site found at 23
Design Notes
The insert was ordered in a pTwist backbone with an AmpR selection marker from Twist Biosciences. The medium copy origin of replication is intended to minimize the metabolic strain on the cells when cotransformed with BBa_K4951070 for switch testing purposes. mStayGold was chosen to serve as a reporter because of its exceptionally high fluorescent signal. The LacZ operon is intended to allow for the switch plasmid to be inducible.
The mStayGold 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.