Difference between revisions of "Part:BBa K4879003"

 
 
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<partinfo>BBa_K4879003 short</partinfo>
 
<partinfo>BBa_K4879003 short</partinfo>
  
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Resistance marker gene for Hygromycin B, designed for expression for <i>Yarrowia lipolytica</i>.
  
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===Usage and Biology===
 
===Usage and Biology===
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Due to the limited number of auxotrophic markers available for <i> Yarrowia lipolytica</i>, we had to use antibiotic-resistance markers for the selection of possible attempted deletion in our chassis. In order to use the selection marker, we had to use 200µg/ml of hygromycin B in our selection plates.
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Hygromycin B tends to kill cells by interfering in polypeptide synthesis, making the organism unable to synthesise proteins. The selection marker gene, which codes for a phosphotransferase, confers resistance by modifying the hygromycin B drug molecule; making it ineffective.
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<html><img src="https://static.igem.wiki/teams/4879/wiki/bba-k4879003-hygr.png"width="250"height="250"></html>
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The protein structure of HygR, as predicted by AlphaFold
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===Design===
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The transcriptional unit for the HygR marker [https://parts.igem.org/Part:BBa_K4879009 (BBa_K4879009)] is designed with the coding sequence flanked by the TEF1 promoter [https://parts.igem.org/Part:BBa_K3629001 (BBa_K3629001)] upstream and the XPR2 terminator [https://parts.igem.org/Part:BBa_K3629004 (BBa_K3629004)] downstream.
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Latest revision as of 07:09, 11 October 2023


Yarrowia lipolytica HygR marker

Resistance marker gene for Hygromycin B, designed for expression for Yarrowia lipolytica.

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Usage and Biology

Due to the limited number of auxotrophic markers available for Yarrowia lipolytica, we had to use antibiotic-resistance markers for the selection of possible attempted deletion in our chassis. In order to use the selection marker, we had to use 200µg/ml of hygromycin B in our selection plates.

Hygromycin B tends to kill cells by interfering in polypeptide synthesis, making the organism unable to synthesise proteins. The selection marker gene, which codes for a phosphotransferase, confers resistance by modifying the hygromycin B drug molecule; making it ineffective.


The protein structure of HygR, as predicted by AlphaFold

Design

The transcriptional unit for the HygR marker (BBa_K4879009) is designed with the coding sequence flanked by the TEF1 promoter (BBa_K3629001) upstream and the XPR2 terminator (BBa_K3629004) downstream.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 153
    Illegal EcoRI site found at 247
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 153
    Illegal EcoRI site found at 247
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 153
    Illegal EcoRI site found at 247
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 153
    Illegal EcoRI site found at 247
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 153
    Illegal EcoRI site found at 247
  • 1000
    COMPATIBLE WITH RFC[1000]