Part:BBa_K3629004

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