Difference between revisions of "Part:BBa K3629004"

(Sequence and Features)
(Sequence and Features)
 
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__TOC__
 
__TOC__
 
===Usage and Biology===
 
===Usage and Biology===
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XPR2 terminator is a native <i>Yarrowia lipolytica</i> terminator for the alkaline XPR2 exonulcease gene. XPR2 is among the most common terminators used for heterologous expression of proteins in <i>Y. lipolytica.</i> (1)
  
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This terminator was used in all of the expression constructs of [https://2020.igem.org/Team:Calgary/Part_Collection our collection.]
  
===Design===
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[https://parts.igem.org/Part:BBa_K3529012 BBa_K3529012]= <i>T. reesei</i> CBHII expression construct <br>
 
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[https://parts.igem.org/Part:BBa_K3529013 BBa_K3529013]= Modified <i>P. funiculosum </i> CBHI expression construct <br>
 
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[https://parts.igem.org/Part:BBa_K3529014 BBa_K3529014]= <i>N. crassa </i> CBHI expression construct <br>
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[https://parts.igem.org/Part:BBa_K3529015 BBa_K3529015]= Nourseothricin resistance expression construct <br>
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[https://parts.igem.org/Part:BBa_K3529016 BBa_K3529016]= Modified <i>T. reesei</i> EGI expression construct <br>
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[https://parts.igem.org/Part:BBa_K3529017 BBa_K3529017]= <i>T. reesei </i> EGII expression construct <br>
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[https://parts.igem.org/Part:BBa_K3529018 BBa_K3529018]= <i> N. patriciarum</i> BGS expression construct <br>
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[https://parts.igem.org/Part:BBa_K3529023 BBa_K3529023]= 2-isopropylmalate synthase (LEU4) overexpression construct <br>
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[https://parts.igem.org/Part:BBa_K3529024 BBa_K3529024]= Anthranilate synthase component 1 (TRP2) overexpression construct <br>
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[https://parts.igem.org/Part:BBa_K3529025 BBa_K3529025]= mCherry expression construct <br>
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[https://parts.igem.org/Part:BBa_K3529026 BBa_K3529026]= mCitrine expression construct <br>
  
 
===Sequence and Features===
 
===Sequence and Features===
 
<partinfo>BBa_K3629004 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3629004 SequenceAndFeatures</partinfo>
 +
 +
There is an EcoRI site within this terminator sequence making this part RFC10 incompatible. However, we added the BioBrick prefix and suffix so that the other enzymes (NotI, XbaI, SpeI, and PstI) can be used to clone this part into an iGEM plasmid or another plasmid. This part can also be cloned through RFC1000 assembly.
  
 
===References===
 
===References===
 
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1. Czajka, Jeffrey J, Nathenson, Justin A, Benites, Veronica T, Baidoo, Edward E. K, Cheng, Qianshun, Wang, Yechun, & Tang, Yinjie J. (2018). Engineering the oleaginous yeast Yarrowia lipolytica to produce the aroma compound β-ionone. Microbial Cell Factories, 17(1), 136–136. https://doi.org/10.1186/s12934-018-0984-x
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Latest revision as of 23:08, 26 October 2020


Yarrowia lipolytica XPR2 terminator

Terminator sequence from the Yarrowia lipolytica alkaline extracellular protease XRP2 gene.


Usage and Biology

XPR2 terminator is a native Yarrowia lipolytica terminator for the alkaline XPR2 exonulcease gene. XPR2 is among the most common terminators used for heterologous expression of proteins in Y. lipolytica. (1)

This terminator was used in all of the expression constructs of our collection.

BBa_K3529012= T. reesei CBHII expression construct
BBa_K3529013= Modified P. funiculosum CBHI expression construct
BBa_K3529014= N. crassa CBHI expression construct
BBa_K3529015= Nourseothricin resistance expression construct
BBa_K3529016= Modified T. reesei EGI expression construct
BBa_K3529017= T. reesei EGII expression construct
BBa_K3529018= N. patriciarum BGS expression construct
BBa_K3529023= 2-isopropylmalate synthase (LEU4) overexpression construct
BBa_K3529024= Anthranilate synthase component 1 (TRP2) overexpression construct
BBa_K3529025= mCherry expression construct
BBa_K3529026= mCitrine expression construct

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 1
    Illegal suffix found in sequence at 271
    Illegal EcoRI site found at 265
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1
    Illegal EcoRI site found at 265
    Illegal SpeI site found at 272
    Illegal PstI site found at 286
    Illegal NotI site found at 7
    Illegal NotI site found at 279
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1
    Illegal EcoRI site found at 265
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 1
    Illegal suffix found in sequence at 272
    Illegal EcoRI site found at 265
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 1
    Illegal EcoRI site found at 265
    Illegal XbaI site found at 16
    Illegal SpeI site found at 272
    Illegal PstI site found at 286
  • 1000
    COMPATIBLE WITH RFC[1000]

There is an EcoRI site within this terminator sequence making this part RFC10 incompatible. However, we added the BioBrick prefix and suffix so that the other enzymes (NotI, XbaI, SpeI, and PstI) can be used to clone this part into an iGEM plasmid or another plasmid. This part can also be cloned through RFC1000 assembly.

References

1. Czajka, Jeffrey J, Nathenson, Justin A, Benites, Veronica T, Baidoo, Edward E. K, Cheng, Qianshun, Wang, Yechun, & Tang, Yinjie J. (2018). Engineering the oleaginous yeast Yarrowia lipolytica to produce the aroma compound β-ionone. Microbial Cell Factories, 17(1), 136–136. https://doi.org/10.1186/s12934-018-0984-x