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Team:Princeton/Parts
From 2008.igem.org
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* '''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] | ||
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==== pLV-TRE-Sox17-Ubc-Bla ==== | ==== pLV-TRE-Sox17-Ubc-Bla ==== | ||
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* '''Description:''' This is the backbone plasmid into which SfiI_Ngn1-linker-EYFP-2A-mKate_SfiI was inserted after digestion with SfiI. | * '''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] | * '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076001 BBa_K076001] | ||
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==== Ngn1 ==== | ==== Ngn1 ==== | ||
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* '''Description:''' Ngn1 is a transcription factor that promotes the differentiation of embryonic stem cells into neuronal precursors. | * '''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] | * '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076002 BBa_K076002] | ||
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==== Linker ==== | ==== Linker ==== | ||
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* '''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. | * '''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] | * '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076003 BBa_K076003] | ||
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==== EYFP ==== | ==== EYFP ==== | ||
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* '''Description:''' This is an enhanced yellow fluorescent reporter protein. | * '''Description:''' This is an enhanced yellow fluorescent reporter protein. | ||
* '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076004 BBa_K076004] | * '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076004 BBa_K076004] | ||
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==== 2A-linker ==== | ==== 2A-linker ==== | ||
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* '''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. | * '''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] | * '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076005 BBa_K076005] | ||
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==== mKate ==== | ==== mKate ==== | ||
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* '''Description:''' This is a red fluorescent reporter protein, found to be more reliable than MCherry or DsRed2. | * '''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] | * '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076006 BBa_K076006] | ||
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==== pLV-TRE-Ngn1-linker-EYFP-Ubc-Bla ==== | ==== pLV-TRE-Ngn1-linker-EYFP-Ubc-Bla ==== | ||
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* '''Description:''' This plasmid only contains the fluorescently-labeled Ngn1, which enables the location of this neuronal transcription factor to be determined within the cell. | * '''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] | * '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076007 BBa_K076007] | ||
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==== D5R or DRD5 ==== | ==== D5R or DRD5 ==== | ||
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* '''Description:''' This gene encodes a specific subunit of a dopamine receptor. | * '''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] | * '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076008 BBa_K076008] | ||
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==== Lbx1 ==== | ==== Lbx1 ==== | ||
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* '''Description:''' This gene codes for a GABAergic inhibitory neurotransmitter. | * '''Description:''' This gene codes for a GABAergic inhibitory neurotransmitter. | ||
* '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076009 BBa_K076009] | * '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076009 BBa_K076009] | ||
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==== GLRA1 ==== | ==== GLRA1 ==== | ||
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* '''Description:''' This gene codes for an inhibitory glycine receptor. | * '''Description:''' This gene codes for an inhibitory glycine receptor. | ||
* '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076010 BBa_K076010] | * '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076010 BBa_K076010] | ||
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==== T2A linker ==== | ==== T2A linker ==== | ||
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* '''Description:''' This part is a protein linker allowing for translational fusion between two gene products. | * '''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] | * '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076011 BBa_K076011] | ||
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==== P2A linker ==== | ==== P2A linker ==== | ||
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* '''Description:''' This part is a protein linker allowing for translational fusion between two gene products. | * '''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] | * '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076012 BBa_K076012] | ||
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==== pLV-TRE-Lbx1-T2A-GLRA1-P2A-D5R-Ubc-Bla ==== | ==== pLV-TRE-Lbx1-T2A-GLRA1-P2A-D5R-Ubc-Bla ==== | ||
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* '''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. | * '''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] | * '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076013 BBa_K076013] | ||
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==== Mash1 ==== | ==== Mash1 ==== | ||
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* '''Description:''' This gene is a cell fate regulator that promotes the differentiation of embryonic stem cells into neuronal precursors. | * '''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] | * '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076014 BBa_K076014] | ||
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==== pLV-TRE-Mash1-IRES2-mKate ==== | ==== pLV-TRE-Mash1-IRES2-mKate ==== | ||
| Line 82: | Line 97: | ||
* '''Description:''' This plasmid encourages differentiation of embryonic stem cells into neuronal precursors and marks the cells with the red-fluorescent 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] | * '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076015 BBa_K076015] | ||
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==== Bleomycin ==== | ==== Bleomycin ==== | ||
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* '''Description:''' This gene is an antibiotic that allows for the selection of cells containing any construct that includes it. | * '''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] | * '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076016 BBa_K076016] | ||
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==== Ubc promoter ==== | ==== Ubc promoter ==== | ||
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* '''Description:''' This is a stable constitutive promoter. | * '''Description:''' This is a stable constitutive promoter. | ||
* '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076017 BBa_K076017] | * '''BioBrick Number:''' [http://partsregistry.org/wiki/index.php?title=Part:BBa_K076017 BBa_K076017] | ||
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==== pLV-TRE-D5R-Ubc-Bleo ==== | ==== pLV-TRE-D5R-Ubc-Bleo ==== | ||
Revision as of 02:18, 30 October 2008
PRINCETON IGEM 2008
| Home | Project Overview | Project Details | Experiments | Results | Notebook |
|---|
| Parts Submitted to the Registry | Modeling | The Team | Gallery |
|---|
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]
