Team:Princeton/Parts

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

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