Difference between revisions of "Part:BBa K1898150"
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<partinfo>BBa_K1898150 short</partinfo>
 
<partinfo>BBa_K1898150 short</partinfo>
  
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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 CH25H (cholesterol 25-Hydroxylase) and a 10x Histidine-tag. CH25H is a catalyst that will make 25 hydroxycholesterol from cholesterol and 10x Histidine-tag is used for protein purification. BBa__B0015 is a double terminator used to stop transcription.  
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===Usage and Biology===
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BBa_K880005 consists of strong promoter and strong rbs to maximize protein production. This construct codes for CH25H (cholesterol 25-Hydroxylase) and a 10x Histidine Tag. CH25H is a catalyst that will make 25 hydroxycholesterol and the 10x Histidine tag is used for protein purification. BBa__B0015 is a double terminator used to stop transcription.  
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==Gel Pictures==
 
  
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PCR was set up after moving the synthesized DNA from IDT to iGEM BIobrick. The expected PCR band sizes are ~1.3 kb.
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===Synthesis Report===
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This construct is synthesized by IDT. The following sequence was sent to IDT to synthesize, and each feature is annotated as shown below:
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*Prefix and suffix are highlighted in orange
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*BBa_K880005 is highlighted in red
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*CH5H is highlighted in light blue
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*10x Histidine is highlighted in green
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*B0015 is highlighted in dark blue
  
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==Sequencing==
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https://static.igem.org/mediawiki/parts/8/8d/Bcht_idt.png
  
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The following sequence was sent to IDT for DNA syntehsis:
 
  
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IDT report shows that the sequence is successfully synthesized:
  
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https://static.igem.org/mediawiki/parts/a/a4/Idt_bcht_1.png
  
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This quality report came back from IDT:
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https://static.igem.org/mediawiki/parts/9/96/Idt_bcht_2.png

Latest revision as of 15:27, 18 October 2016


Strong promoter + Strong RBS + CH25H + 10x His tag + Double terminator

BBa_K880005 consists of a strong promoter and strong RBS and was used to maximize protein production. This construct codes for CH25H (cholesterol 25-Hydroxylase) and a 10x Histidine-tag. CH25H is a catalyst that will make 25 hydroxycholesterol from cholesterol and 10x Histidine-tag is used for protein purification. BBa__B0015 is a double terminator used to stop transcription.

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
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]



Synthesis Report

This construct is synthesized by IDT. The following sequence was sent to IDT to synthesize, and each feature is annotated as shown below:

  • Prefix and suffix are highlighted in orange
  • BBa_K880005 is highlighted in red
  • CH5H is highlighted in light blue
  • 10x Histidine is highlighted in green
  • B0015 is highlighted in dark blue

Bcht_idt.png


IDT report shows that the sequence is successfully synthesized:

Idt_bcht_1.png

Idt_bcht_2.png