Difference between revisions of "Part:BBa K3697010"

 
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<partinfo>BBa_K3697010 short</partinfo>
 
<partinfo>BBa_K3697010 short</partinfo>
  
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This part is an expression vector for Bacillus subtilis that produces mCherry_BSU, a codon-optimized mCherry for expression in B. subtilis, for integration into Bacillus subtilis at the AmyE loci. mCherry_BSU is under pVeg expression, the strongest constitutive promotor known in B. subtilis. Plasmid expresses kanamycin resistance in B. subtilis, and ampicillin resistance for cloning in E. coli.
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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
 
mCherry_BSU has an excitation peak at 585 nm and a peak emission at 615 nm
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===Usage and Biology===
 
===Usage and Biology===
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This mCherry was codon optimized for B. subtilis expression.
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Due to the high levels of mCherry expression from pVeg, this vector can be used to signal transformation, transcription, and translation in B. subtilis.
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Revision as of 23:58, 23 October 2020


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.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE 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
  • 12
    INCOMPATIBLE 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
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4308
    Illegal BglII site found at 1585
    Illegal XhoI site found at 1218
  • 23
    INCOMPATIBLE 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
  • 25
    INCOMPATIBLE 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
  • 1000
    INCOMPATIBLE 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