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Team:Princeton/Parts
From 2008.igem.org
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==== pLV-TRE-Ngn1-linker-EYFP-2A-mKate-Ubc-Bla ==== | ==== 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. | + | * ''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] | + | * ''BioBrick Number:'' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076000 BBa_K076000] |
Revision as of 02:10, 30 October 2008
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]
