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Team:Princeton/Parts
From 2008.igem.org
PRINCETON IGEM 2008
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Coding Parts Submitted to the Registry
Ubc promoter
- Description: This is a stable constitutive promoter.
- BioBrick Number: BBa_K076017
Ngn1
- Description: This is a transcription factor that promotes the differentiation of embryonic stem cells into neuronal precursors.
- BioBrick Number: BBa_K076002
Mash1
- Description: This gene is a cell fate regulator that promotes the differentiation of embryonic stem cells into neuronal precursors.
- BioBrick Number: BBa_K076014
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: BBa_K076003
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: BBa_K076005
T2A linker
- Description: This is a protein linker allowing for translational fusion between two gene products.
- BioBrick Number: BBa_K076011
P2A linker
- Description: This is a protein linker allowing for translational fusion between two gene products.
- BioBrick Number: BBa_K076012
mKate
- Description: This is a red fluorescent reporter protein, found to be more reliable than mCherry or DsRed2.
- BioBrick Number: BBa_K076006
EYFP
- Description: This is an enhanced yellow fluorescent reporter protein.
- BioBrick Number: BBa_K076004
D5R or DRD5
- Description: This gene encodes a specific subunit of a dopamine receptor.
- BioBrick Number: BBa_K076008
Lbx1
- Description: This gene codes for a GABAergic inhibitory neurotransmitter.
- BioBrick Number: BBa_K076009
GLRA1
- Description: This gene codes for an inhibitory glycine receptor.
- BioBrick Number: BBa_K076010
Bleomycin
- Description: This gene is an antibiotic that allows for the selection of cells containing any construct that includes it.
- BioBrick Number: BBa_K076016
Plasmids Submitted to the Registry
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: BBa_K076015
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: BBa_K076007
pLV-TRE-Ngn1-linker-EYFP-2A-mKate-Ubc-Bla
- Description: This plasmid allows rtTA-induced expression of the cell-fate regulator Ngn1 linked to an enhanced yellow fluorescent protein (EYFP) reporter, and a red fluorescent protein (mKate) 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: 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: BBa_K076001
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: BBa_K076018
pLV-TRE-Lbx1-T2A-GLRA1-P2A-D5R-Ubc-Bla
- Description: This plasmid allows for the simultaneous production of Lbx1 [produces GABAergic neurons], GLRA1 [produces a specific glycine receptor], and D5R [produced 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: BBa_K076013
