Part:BBa_K731700:Design
Platform for terminators analysis under the control of T7 promoter
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 6850
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NotI site found at 9
Illegal NotI site found at 6856 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 6850
Illegal BglII site found at 6011
Illegal BamHI site found at 6832
Illegal XhoI site found at 753 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found at 6850
Illegal suffix found at 2 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found at 6850
Plasmid lacks a suffix.
Illegal XbaI site found at 6865
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NgoMIV site found at 714
Illegal NgoMIV site found at 1051
Illegal NgoMIV site found at 4231
Illegal NgoMIV site found at 4391
Illegal NgoMIV site found at 5979
Illegal AgeI site found at 6800 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal BsaI site found at 2228
Illegal SapI.rc site found at 3310
Design Notes
Using and modifying this construct is important to remember that the two proteins have strong N-terminal homology, especially designing primers. For any future improvement, maybe it would be better two use two different fluorescence proteins, because these two has partially overlapping emission spectra and different folding kinetics. Using and modifying this construct is important to remember that the two proteins have strong N-terminal homology, especially designing primers. For any future improvement, maybe it would be better two use two different fluorescence proteins, because these two has partially overlapping emission spectra and different folding kinetics.
Source
It's made from a Roberta Lentini's construct (from Mansy lab) that consist of pET21b with the two proteins and a 20bp linker between them. That construct was mutated two times to eliminate illegal restriction sites; prefix-suffix linker was added by insertion PCR.