Team:Princeton/Parts

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PRINCETON IGEM 2008

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Parts Submitted to the Registry

pLV-TRE-Ngn1-linker-EYFP-2A-mKate-Ubc-Bla

  • Description: A plasmid allowing rtTA-induced expression of the cell-fate regulator Ngn1 linked to an enhanced yellow fluorescent protein reporter, and a red fluorescent protein reporter tethered by a 2A linker. This construct allows for the verification of the 2A linker, with the ideal result of yellow-labeled Ngn1 remaining in the nucleus and red-labeled mKate being exported to the cytoplasm.
  • BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076000 BBa_K076000]


pLV-TRE-Sox17-Ubc-Bla

  • Description: This is the backbone plasmid into which SfiI_Ngn1-linker-EYFP-2A-mKate_SfiI was inserted after digestion with SfiI.
  • BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076001 BBa_K076001]


Ngn1

  • Description: Ngn1 is a transcription factor that promotes the differentiation of embryonic stem cells into neuronal precursors.
  • BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076002 BBa_K076002]


Linker

  • Description: This short sequence connects the Ngn1 and EYFP gene products, providing a translational fusion. This enables us to detect the location of Ngn1 by tracking the appearance of fluorescent yellow protein within the cell.
  • BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076003 BBa_K076003]


EYFP

  • Description: This is an enhanced yellow fluorescent reporter protein.
  • BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076004 BBa_K076004]


2A-linker

  • Description: This is a protein linker sequence that allows one reporter protein, in our case EYFP, to be linked to another, in our case mKate, under certain conditions and cleaved under certain other conditions.
  • BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076005 BBa_K076005]


mKate

  • Description: This is a red fluorescent reporter protein, found to be more reliable than MCherry or DsRed2.
  • BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076006 BBa_K076006]


pLV-TRE-Ngn1-linker-EYFP-Ubc-Bla

  • Description: This plasmid only contains the fluorescently-labeled Ngn1, which enables the location of this neuronal transcription factor to be determined within the cell.
  • BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076007 BBa_K076007]


D5R or DRD5

  • Description: This gene encodes a specific subunit of a dopamine receptor.
  • BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076008 BBa_K076008]


Lbx1

  • Description: This gene codes for a GABAergic inhibitory neurotransmitter.
  • BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076009 BBa_K076009]


GLRA1

  • Description: This gene codes for an inhibitory glycine receptor.
  • BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076010 BBa_K076010]


T2A linker

  • Description: This part is a protein linker allowing for translational fusion between two gene products.
  • BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076011 BBa_K076011]


P2A linker

  • Description: This part is a protein linker allowing for translational fusion between two gene products.
  • BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076012 BBa_K076012]


pLV-TRE-Lbx1-T2A-GLRA1-P2A-D5R-Ubc-Bla

  • Description: This plasmid allows for the simultaneous production of Lbx1 [produces GABAergic neurons], GLRA1 [a glycine receptor], and D5R [a subunit of a particular dopamine receptor]. This makes the neurons able to respond to both a dopamine and a glycine input, and in turn produce a GABA output.
  • BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076013 BBa_K076013]


Mash1

  • Description: This gene is a cell fate regulator that promotes the differentiation of embryonic stem cells into neuronal precursors.
  • BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076014 BBa_K076014]


pLV-TRE-Mash1-IRES2-mKate

  • Description: This plasmid encourages differentiation of embryonic stem cells into neuronal precursors and marks the cells with the red-fluorescent mKate.
  • BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076015 BBa_K076015]


Bleomycin

  • Description: This gene is an antibiotic that allows for the selection of cells containing any construct that includes it.
  • BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076016 BBa_K076016]


Ubc promoter

  • Description: This is a stable constitutive promoter.
  • BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076017 BBa_K076017]


pLV-TRE-D5R-Ubc-Bleo

  • Description: This plasmid allows bleomycin-selected cells to express the D5R dopamine receptor subunit upon induction with doxycycline.
  • BioBrick Number: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076018 BBa_K076018]
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