Part:BBa_K3697010
mCherry_BSU Plasmid
This part is an expression vector for Bacillus subtilis that produces codon-optimized mCherry_BSU, for integration into Bacillus subtilis at the AmyE loci (BBa_K143001). mCherry_BSU is under pVeg expression (BBa_K143012), the strongest constitutive promotor known in B. subtilis. Plasmid expresses kanamycin resistance in B. subtilis, and ampicillin resistance for cloning in E. coli.
mCherry_BSU has an excitation peak at 585 nm and a peak emission at 615 nm
This plasmid successfully integrates into B. subtilis and produces mCherry_BSU at levels that can be quantified using a fluorescent plate reader. Transformation was performed using xylose inducible competent B. subtilis, and plated on selective kanamycin media (20ug/mL). When cloning in E. coli, the E. coli may be red, as they will non-discriminately express the mCherry_BSU. E. coli transformed with this plasmid have been clearly red under natural light for the duration of their lifetime and does not appear to degrade.
The RBS for expression of mCherry in this vector is strong.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 4308
Illegal XbaI site found at 1568
Illegal SpeI site found at 1472
Illegal PstI site found at 117
Illegal PstI site found at 1713
Illegal PstI site found at 3862
Illegal PstI site found at 4314 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 4308
Illegal SpeI site found at 1472
Illegal PstI site found at 117
Illegal PstI site found at 1713
Illegal PstI site found at 3862
Illegal PstI site found at 4314 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 4308
Illegal BglII site found at 1585
Illegal XhoI site found at 1218 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 4308
Illegal XbaI site found at 1568
Illegal SpeI site found at 1472
Illegal PstI site found at 117
Illegal PstI site found at 1713
Illegal PstI site found at 3862
Illegal PstI site found at 4314 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 4308
Illegal XbaI site found at 1568
Illegal SpeI site found at 1472
Illegal PstI site found at 117
Illegal PstI site found at 1713
Illegal PstI site found at 3862
Illegal PstI site found at 4314
Illegal NgoMIV site found at 1254
Illegal AgeI site found at 3532 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 291
Illegal BsaI site found at 5940
Illegal BsaI site found at 5972
Illegal BsaI.rc site found at 5928
Illegal BsaI.rc site found at 5960
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