30C12HSL input, LuxAHL output (luxI is LVA+)
glows red when its hot as promoter no longer repressed by cIts. This variant has strong promoter and RBS to compensate for weak mRFP1.
Threshold: Activation module (IS10-type cI antisense)
Part of the thresholding device (works with j07036 and j07037). This encodes the antisense mRNA to the cI transcript (j07036), and hence regulates cI in the cell and thereby indirectly controls the level of expression of reporter gene (j07037). This part encodes antisense mRNA in the same way as IS10 transposase is regulated by IS10 gene.
Threshold: Repression module (cI repressor with junk region)
Part of the thresholding device (works with j07037, j07038/j07039/j07040). This encodes the cI repressor that normally represses j07037. However, j07038/39/40 code for antisense mRNA to this, which, when transcribed, represses translation of cI and activates j07037
Lux AHL input, 30C12HSL output (lasI is LVA+)
Production of GFP is at high temperartures, when CI(ts) is degraded.
This is part of a two component color based temperature reporter, similar to an liquid crystal thermometer (commonly used for fishtanks, etc). Not active at 37C when lacIts protein is functional, GFP is not produced. As the temperature rises, the promoter becomes active and GFP is made.
Part of system construction Includes strong RBS (B0030) and FecI (J07022)
No fluorescence observed. J06701 (mCherry with RBS and T) is bad.
variant of J11005 (low temp color) with stronger RBS
This part includes strong RBS (B0030), and FecR' (J07023). Intermediate for test construct.
Fred Tan, Debra Lin, Jay Blumling, Madeleine Sheldon-dante, MIT SMUG
FecI test construct w/ inducible promoter
Test construct for FecI Inducible promoter (regulated by LacI) + strong RBS (B0030) + FecI (J07022)
This part will be used as a readout for the promoter that naturally promotes production of FecA, and is regulated by FecI. See J07019 for more details.
TEST [pLam cI -][str RBS][GFP][dbl Terminator]
constitutive expressioin of tetR protein
Threshold: Activation module (cI anti-sense)
Part of the thresholding device (works with j07036 and j07037). This encodes the antisense mRNA to the cI transcript (j07036), and hence regulates cI in the cell and thereby indirectly controls the level of expression of reporter gene (j07037). This part encodes simply for the antisense mRNA of j07036 (KISS method).
This part results from the ligation of E0040 with B0015 in the manner described on this website.
YFP on LuxBox, no terminator [Construction Intermediate]
Tet-repressible production of CI(ts) to allow tuning of temperature readouts
LVA- version of J06201. LVA- protein coding region not a Biobrick
YFP on LasBox, no terminator [Construction Intermediate]
Antiswitch repressing Biobrick coding sequence at the RBS ("Off" switch)
The presence of theophylline prevents translation of mRNA coding region directly downstream from any Biobrick that contains the BBa_B0034 RBS. This effect is the result of the antiswitch attaching to the B0034 RBS preceding the coding region. The absence of theophylline allows expression of the coding region as usual.
Transcription of the antiswitch is repressed in the presence of tetR. Also, the actual RBS sequence of BBa_B0034 is only 12 bp long; three additional base pairs comprise the total target sequence, and these additional 3 bp's correspond to the last three base pairs of the B0034 Biobrick Prefix, Xba I.
Adaptation of antiswitch design "S1" by Bayer and Smolke; article available at http://www.nature.com/nbt/journal/v23/n3/full/nbt1069.html
Bacterial Whiteboard by Hannah (w/ Minimal Available Parts)
When lactose is added to the system, bind to lac repressor (endogenous) therefore CFP (fast degrading) produced - glow cyan. When Tet added to the system, drive transcription of lacZ beta galactosidase which hydrolyses latose and lac repressor active. Stop Glowing cyan. ***NOTE*** This system will ONLY WORK IN LAC Z- bugs!!!!!
This part includes strong RBS (B0030), FecR' (J07023), and strong terminator (B0015). Built as an intermediate part of overall construct.
Outputs YFP when given 30C12HSL (YFP changed from E0434 to E0432, 6/27)
ToxR wt [optimized (without EcoRI site)]
J06700 (Venus YFP with RBS and T) is bad.
No fluorescence observed. J06700 (Venus YFP with RBS and T) is bad.
temperature sensor - low temperature color
Provides low temperature (37C) color signal (red) in presence of CI or CI(ts).
CI protein activates the production of RFP, but cannot repress. The CI(ts) protein becomes inactive at higher temperatures and is used in trans with another color reporter.
variant of J11005 uncoupled from CI(ts) production
low temperature RFP expression uncoupled from CI(ts) protein
TEST [OnLock2][cI][DblTerminator][pRM GFP]
testing ability of lock 2 in context of f-mobilizable plasmid
Intermediate in two promoter screening plasmid
TEST [OriTF][pLam cI -][str RBS][GFP][dbl Terminator]
Threshold: Activation module (cI antisense with hairpins)
Part of the thresholding device (works with j07036 and j07037). This encodes the antisense mRNA to the cI transcript (j07036), and hence regulates cI in the cell and thereby indirectly controls the level of expression of reporter gene (j07037). This part encodes antisense mRNA of j07036 along with hairpin loops on the edges to keep it linear.
Lambda promotor fused with zinc finger binding site for repression
This part results from ligating E1010 with B0015 in the manner described on this website.
ToxR w.t. (from Vibrio Cholarae, aa's 1-294)
ToxR is a dimerizing transmembrane protein with dna binding site. mit_igem_2005 implementing to develop a cellular sensing platform that accomodates an array of choosable antigens. thorough story @ http://info.bio.cmu.edu/Courses/03441/TermPapers/99TermPapers/ChoTox/coreg.html
temporary (DE -- delete at some future point as needed)
fim E inverts fim S "on" (BBa_T4003) to "off" (BBa_T4004). Fim E binds to the inverted repeats of fim S.
variant of J11004 (hi temp color) with weaker RBS
Variant of BBa_J1104 with weaker RBS. Concern is that GFP signal will be too strong. The expression of GFP should not start until 37C, but a weaker promoter will help ensure this.
FecR' test construct w/ inducible promoter
This part is a test construct and measurement for FecR'. It includes: inducible promoter (R0010), strong RBS (B0030), FecR' (J07023), and GFP (E0840).
Built by Josh as an intermediate in screening plasmid construction.
Front side of Tox_cell_signaling_by_dimerization_system. Recievers to be attached downstream of the Tox protein with intent to induce controlled dimerization of Tox. System used in conjunction with J07007 set of ctx_promotor::[reaction] parts.
FecI test construct w/ constitutive promoter
Test construct for FecI Constitutive promoter (R0040) regulated by TetR protein + strong RBS (B0030) + FecI (J07022)
Consensus sequence for highly expressions eukaryotic proteins
YFP on RhlBox, no terminator [Construction Intermediate]
Glows green when cool. cIts activates promoter but dissociates and removes activation when hot. Based obn J11009.