Difference between revisions of "Part:BBa K5103001"

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Streak plate of PCG-Cry8Da transformant colonies grown on ampicillin selective media.  
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Figure 1. Streak plate of PCG-Cry8Da transformant colonies grown on ampicillin selective media.  
  
 
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PCG-CRY plasmid components were developed using SnapGene.
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Figure 2. PCG-CRY plasmid components were developed using SnapGene.
  
 
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Revision as of 21:52, 28 September 2024


PCG004 with Cry8Da

It includes features for selective gene expression in a dropout region that normally contains green fluorescent protein (GFP) flanked by BsaI sites. In this case we replaced GFP with Cry8Da crystal toxin sequences from Bacillus thuringiensis (Bt). Upstream, a lacI sequence encodes the lac repressor and a lac operator (lacO) overlaps with the pGrac promoter. Using this plasmid allows us to induce the expression of Cry8Da when we want using IPTG. This allows transcription of Cry8Da by causing a conformational change in Lac, releasing it from lacO.

Figure 1. Streak plate of PCG-Cry8Da transformant colonies grown on ampicillin selective media.

Figure 2. PCG-CRY plasmid components were developed using SnapGene.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 595
    Illegal EcoRI site found at 6398
    Illegal EcoRI site found at 8948
    Illegal EcoRI site found at 9027
    Illegal EcoRI site found at 10439
    Illegal EcoRI site found at 11102
    Illegal XbaI site found at 11533
    Illegal PstI site found at 8079
    Illegal PstI site found at 9520
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 595
    Illegal EcoRI site found at 6398
    Illegal EcoRI site found at 8948
    Illegal EcoRI site found at 9027
    Illegal EcoRI site found at 10439
    Illegal EcoRI site found at 11102
    Illegal NheI site found at 6234
    Illegal PstI site found at 8079
    Illegal PstI site found at 9520
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 595
    Illegal EcoRI site found at 6398
    Illegal EcoRI site found at 8948
    Illegal EcoRI site found at 9027
    Illegal EcoRI site found at 10439
    Illegal EcoRI site found at 11102
    Illegal BglII site found at 66
    Illegal BglII site found at 3147
    Illegal XhoI site found at 3151
    Illegal XhoI site found at 11410
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 595
    Illegal EcoRI site found at 6398
    Illegal EcoRI site found at 8948
    Illegal EcoRI site found at 9027
    Illegal EcoRI site found at 10439
    Illegal EcoRI site found at 11102
    Illegal XbaI site found at 11533
    Illegal PstI site found at 8079
    Illegal PstI site found at 9520
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 595
    Illegal EcoRI site found at 6398
    Illegal EcoRI site found at 8948
    Illegal EcoRI site found at 9027
    Illegal EcoRI site found at 10439
    Illegal EcoRI site found at 11102
    Illegal XbaI site found at 11533
    Illegal PstI site found at 8079
    Illegal PstI site found at 9520
    Illegal AgeI site found at 9294
    Illegal AgeI site found at 10176
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 11524
    Illegal BsaI.rc site found at 7949
    Illegal BsaI.rc site found at 7962
    Illegal BsaI.rc site found at 11537
    Illegal SapI.rc site found at 1219
    Illegal SapI.rc site found at 5124