Difference between revisions of "Part:BBa K1898450"

 
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BBa_K880005 is used to provide a strong promoter and strong rbs to maximize protein output. The ORF includes cryab, a crystallin protein exists in the lens and other tissues, and 10x Histidine tag for protein purification. BBa_B0015 consists of a double terminator to end transcription.
 
BBa_K880005 is used to provide a strong promoter and strong rbs to maximize protein output. The ORF includes cryab, a crystallin protein exists in the lens and other tissues, and 10x Histidine tag for protein purification. BBa_B0015 consists of a double terminator to end transcription.
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BBa_K880005 consists of a strong promoter and strong RBS and was used to maximize protein production. This construct codes for CRYAB (crystallin alpha B) and a 10x Histidine-tag. CRYAB is a crystallin protein abundant in the lens and 10x Histidine-tag is used for protein purification. BBa__B0015 is a double terminator used to stop transcription.
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For our project, this construct allows us to 1) test our protein purification method and 2) see the effect of oxidative stress caused by the addition of reactive oxygen species 3) analyze the effectiveness of our prevention and treatment.
  
  
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<partinfo>BBa_K1898450 parameters</partinfo>
 
<partinfo>BBa_K1898450 parameters</partinfo>
 
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==Sequencing==
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After running a PCR check and saw the band at the expected size (~1kb), we sent the DNA out for sequencing. The sequencing result is shown below.
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* The four restriction sites are highlighted in red
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* BBa_K880005 is highlighted in light blue
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* CRYAB is highlighted in orange
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* 10x Histidine-tag is highlighted in green
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* BBa_B0015 is highlighted in dark blue
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https://static.igem.org/mediawiki/parts/8/82/Bbht_vf2.png

Revision as of 03:02, 19 October 2016


Strong promoter + Strong RBS + CRYAB + 10x Histidine tag + Double terminator

BBa_K880005 is used to provide a strong promoter and strong rbs to maximize protein output. The ORF includes cryab, a crystallin protein exists in the lens and other tissues, and 10x Histidine tag for protein purification. BBa_B0015 consists of a double terminator to end transcription.

BBa_K880005 consists of a strong promoter and strong RBS and was used to maximize protein production. This construct codes for CRYAB (crystallin alpha B) and a 10x Histidine-tag. CRYAB is a crystallin protein abundant in the lens and 10x Histidine-tag is used for protein purification. BBa__B0015 is a double terminator used to stop transcription.

For our project, this construct allows us to 1) test our protein purification method and 2) see the effect of oxidative stress caused by the addition of reactive oxygen species 3) analyze the effectiveness of our prevention and treatment.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 87
    Illegal BamHI site found at 429
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 514
    Illegal SapI.rc site found at 374



Sequencing

After running a PCR check and saw the band at the expected size (~1kb), we sent the DNA out for sequencing. The sequencing result is shown below.

  • The four restriction sites are highlighted in red
  • BBa_K880005 is highlighted in light blue
  • CRYAB is highlighted in orange
  • 10x Histidine-tag is highlighted in green
  • BBa_B0015 is highlighted in dark blue

Bbht_vf2.png