Part:BBa_K731700
Platform for terminators analysis under the control of T7 promoter
We used this part to test terminators: the two subsequent proteins allow termination efficiency's quantification as the ratio between Venus's and Cherry's fluorescence. The vector is pET21b, that contains lacI, t7 promoter and ampicillin resistance gene.We inserted the prefix-suffix linker between the two proteins, to allow terminator's characterization, and mutated the two illegal sites present in the original plasmid.
The two fluorescent proteins have partially overlapping emission spectra, thus, to avoid unwanted interferences, the fluorimetric measures should be taken with these parameters: Venus excitation: 485nm, Venus emission: 528nm Cherry excitation: 528nm, Cherry emission: 615nm.
Sequence and Features
- 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
//plasmid/measurement
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