Parts Shopping List
From 2006.igem.org
Composite bricks - these are the best to use as less cloning is required.
All these bricks express YFP and downregulate it in response to a signal
YELLOW
ARABINOSE RESPONSE.
BBa_E0610: Arabinose turns on lac repressor which in turn shuts down Yellow fluorescent protein.
BBa_E0600: like BBa_E0610 but YFP is destabilized - slow degradation. Not listed as available.
BBa_E0611: Arabinose turns on destabilized TetR repressor which shuts down yellow fluorescent protein. Addition of Tetracyline blocks TetR repressor allowing yellow fluorescent protein to be turned back on.
BBa_E0601: Like BBa_E0611 but YFP is destabilized - slow degradation. Not listed as available
BBa_E0613: Arabinose turns on destabilized lambda repressor which shuts down yellow fluorescent protein.
BBa_E0602: Like BBa_E0613 but YFP is destabilized - slow degradation. Not listed as available
BBa_E0603: Like BBa_E0602 but baseline expression of YFP is higher. Not listed as available
BBa_E0604: Arabinose turns on destabilized 434 cl epressor which in turn shuts down destabilized (slow) Yellow fluorescent protein. Not listed as available
BBa_E0614: Like BBa_E0604 but YFP is not destabilized (hangs around forever). Not listed as available
BBa_E0605: Arabinose turns on destabilized p22 repressor which in turn shuts down destabilized (slow) Yellow fluorescent protein. Not listed as available
BBa_E0615: Like BBa_E0605, but YFP is not destabilized, hangs around forever.
Lac I REGULATED
BBa_I12026: Produces YFP (not destabilized). Controlled by Lac I BBa_I14022: Slow-destabilized (LVA) YFP controlled by Lac promoter
MULTIPLE REGULATORS
BBa_I12051: Lactose induces lambda repressor. Arabinose induces 434 repressor. Lambda repressor activates YFP, 434 represses YFP. YFP is destabilized (AAV, fast).
BBa_I12052: Like BBa_I12051 but promoter for YFP has double binding sites for better control.
LAMBDA regulated
BBa_I13973: Fast destabilized (AAV) YFP controlled by original lambda promoter - lambda protein would repress it.
LAS regulated
BBa_I14013: Slow destabilized YFP under Las promoter.
BBa_I14030: TetR promoter controls LacI repressor, lambda repressor, and YFP. All three are slow (LVA) destabilized.
BASIC COLOR LEGO
BBa_I14005: Slow-destabilized (LVA) YFP with strong ribosome binding site
CYAN
TET response
BBa_I0401: Expresses CFP and lux from the Tet-OFF promoter. Neither is destabilized. Not listed as available
BBa_I0402: Like BBa_I0401 but lux is destabilized (LVA, slow)
BBa_I0404: Produces rhlR and CFP, downregulated by Tet. rhlR binds with N-butyryl-HSL, regulates transcription. rhlR but not CFP is destabilized.
BBa_I0405: same as BBa_I0404, but produces rhIl which produces N-butyryl-HSL.
BBa_I0406: produces CFP and lasR, which accepts chemical signal AI-1. Downregulated by Tet. LasR but not CFP are destabilized.
BBa_I0407. Like BBa_I0406 but produces lasI (destabilized LVA slow) which generates the chemical signal Al-1.
BBa_I0408: produces CFP and CinR, repressed by Tetracycline. Cin R is LVA (slow) destabilized. Cin R binds O3-C14:1-HSL and activates the Cin promoter.
BBa_I0409: Like BBa_I0408 but produces cinL (destabilized LVA slow) which generates the chemical signal O3-C14:1-HSL.
BBa_I14029: LacI promoter controls slow destabilized (LVA) TetR and slow destabilized (LVA) YFP
ARABINOSE response:
BBa_I0403 Expresses CFP and aiiA from an arabinose promoter. aiiA degrades N-acyl homoserine lactones (quorum sensing autoinducers). aiiA but not CFP is destabilized (LVA, slow).
GREEN
ARABINOSE
BBa_I13540: Arabinose turns on green fluorescent protein.
RED:
BBa_I13507: Basic red fluorescent protein: ribosome binding site, red, stop. B.Y.O. promoter.
ARABINOSE
BBa_I13520: Arabinose turns on mRFP. This think was actually shown to work. BBa_I13517: Like BBa_I13520 but weaker ribosome binding site.
SIGNALING
Promoterless (BYOP)
BBa_I0460 aiiA cassette. Produces enzyme which degrades N-acyl homoserine lactones (quorum sensing autoinducers. Destabilized with LVA (slow) B.Y.O.P (Bring your own promoter).
BBa_I10100: Produces adhesin, makes bacteria stick to each other. B.Y.O.P (Bring your own promoter)
BBa_I14002: LVA-destabilized Lac I, with strong ribosome binding site.
BBa_I14003: LVA-destabilized TetR, with strong ribosome binding site.
BBa_I14010: LVA-destabilized Ara C, with strong ribosome binding site.
BBa_I14012: LVA-destabilized LasR, with strong ribosome binding site.
BBa_I14045: LVA-destabilized RhiR, with strong ribosome binding site
BBa_I14046: LVA-destabilized RhiR, with strong ribosome binding site
With promoter
BBa_I14023: Constitutively expresses slow-destabilized LasR protein. BBa_I14023: Constitutively expresses slow-destabilized RhiR protein.
BBa_I12002: Lambda promoter driving expression of 434 repressor. Not destabilized. BBa_I12054: same as BBa_I12002, lambda promoter is modified to be activated by lac repressor cI
BBa_I12055: Hybrid lambda promoter activated by lambda and repressed by 434, driving expression of destablized lambda (LVA slow degradation). This was used as part of an oscillator. BBa_I12056: Like BBa_I12055:but the promoter has duplicate sites for better control.
BBa_I12011: Lambda promoter driving expression of p22 repressor. LVA (slow) destabilized.
BBa_I13271: Cassette: rhiR upstream of rhi promoter. rhiR with butyl-homoserine lactone activates rhi promoter. rhiR needs a promoter, not included. BBa_I13303: Like BBa_I13271 but controlled by lac promoter. BBa_I13309: like BBa_I13303 BBa_I13310: like BBa_I13303 but controlled by TetR BBa_I13312: like BBa_I13303 but controlled by TetR
BBa_I14005, BBa_I14038: RhI promoter upstream of Lac I
BBa_I13305: Lac I induces LasR, LasR with homoserine lactone activates Las promoter (downstream). Can add your own gene after Las promoter.
BBa_I14014: slow destabilized (LVA) LasR under Lac I promoter
BBa_I13016, BBa_S04003: Arabinose induces LVA (slow) destabilized TetR protein.
BBa_I13017, BBa_S04002: Expression of LVA(slow) destabilized TetR protein controlled by lactose (Lac I promoter).
BBa_I14028, BBa_I14036: LVA-destabilized Lambda controlled by LasR promoter.
BBa_I14037: Destabilized LasL controlled by LasR promoter. Lambda operator allows shutdown by lambda repressor.
BBa_I14050: Destabilized RhiL upregulated by Las/HSL and blocked by TetT
BBa_J06204: Makes lambda cl protein in response to C4-HSL
CHAIN REACTION PREMADE:
BBa_I13036: Arabinose induces TetR. TetR induces LuxR. LuxR induces YFP if a homoserine lactone signal is received.
BBa_I13037: Lactose induces TetR. TetR induces LuxR. Arabinose induces LuxL which produces homoserine lactone. LuxR and homoserine lactone activate YFP.
BBa_I13038: Arabinose induces TetR, and also luxL which makes homoserine lactone. TetR induces LuxR. LuxL and LuxR together activate YFP.
BBa_I13920: Lux pL makes Lux R protein. Lux R protein + homoserine lactone shuts off Lux pL (no more lux R) and activates Lux pR which makes LacI. Lac I shuts off luxL protein (homoserine lactone) which shuts down lac I and HSL production, Lux pL is free to start over. Lac I and LuxL are slow destabilized, LVA.
BBa_I13921: Same as BBa_I13920 but TetR (LVA destabilized) is substituted for Lac I.
BBa_I13922: Same as BBa_I13920 but lambda repressor (LVA destabilized) is substituted for Lac I.
BBa_J06910: Lambda induces destabilized (slow LVA) LacI. LacI shuts down EYFP (nondestabilized).
BBa_J06913: If you add 434, you repress lambda (LVA slow destabilized). YFP is downstream of a lambda promoter so by adding 434 you reduce repression and turn on YFP.
MOTILITY
TetR control
BBa_I13710: TetR induces CheY, which is a phosphorylation target for motility.
BBa_I13711: TetR induces CheB. CheB can be phosphorylated and will demethylate Aspartate receptors.
BBa_I13712: TetR induces CheR. CheR opposes Che B.
LACTOSE control
BBa_I13721: Lactose induces CheB. (lac promoter)
MULTI-COMPONENT
BBa_I13730: Lac I controls Che B, TetR controls Che R. Che B opposes Che R (chemosensing)
BBa_M30109: light sensor. Activates OmpC and Represses Omp F promoter
BBa_J11002: Generates GFP pulse, slow, not destabilized.
BBa_J06917: Lambda induces 434, which represses YFP. not destabilized.
BASIC TOOLS
BBa_I13800: ampicillin resistance
BBa_I13801: kanamycin resistance
SIMPLE COLOUR BLOCKS
CYAN BBa_E0420 ECFP (RBS+ LVA- TERM): (B0034.E0020.B0015) BBa_I15012 ECFP (RBS+ LVA-): (B0030.E0020) BBa_I15016 ECFP (RBS+ LVA-): (B0032.E0020) BBa_I15019 ECFP (RBS+ LVA-): (B0031.E0020) BBa_I13600 ECFP (tetR promo RBS+ LVA- TERM): (R0040.E0420) BBa_I13601 ECFP (lacI/pL promo RBS+ LVA- TERM): (R0011.E0420) BBa_E0422 ECFP (RBS+ LVA+ TERM): (B0034.E0022.B0015) BBa_J04421 ECFP (LacI promo RBS+ LVA+ TERM): (R0010.E0422) BBa_I13602 ECFP (tetR promo RBS+ LVA+ TERM): (R0040.E0422), in absence of tetR ECFP is expressed BBa_I13603 ECFP (lacI/pL promo RBS+ LVA+ TERM): (R0011.E0422) BBa_E0024 ECFP (RBS- AAV+): more stable than E0022 but less than E0026? BBa_E0424 ECFP (RBS+ AAV+ TERM): (B0034.E0024.B0015) not listed as available BBa_E0026 ECFP (RBS- ASV+) BBa_E0426 ECFP (RBS+ ASV+ TERM): (B0034.E0026.B0015) not listed as available BBa_S03474: lambda represses CFP. Has RBS + TERM. Has double lambda sites. BBa_S03465: lambda activates CFP. Has RBS + TERM
YELLOW BBa_E0430 EYFP (RBS+ LVA- TERM): (B0034.E0030.B0015) BBa_I13001 EYFP (RBS+ LVA- TERM): (B0030.E0030.B0015) strong rbs BBa_I13002 EYFP (RBS+ LVA- TERM): (B0031.E0030.B0015) BBa_I13003 EYFP (RBS+ LVA- TERM): (B0032.E0030.B0015) BBa_I13004 EYFP (RBS+ LVA- TERM): (B0033.E0030.B0015) BBa_E0438 EYFP (RBS+ LVA+ TERM): (B0043.E0032.B0014): difficult to detect fluorescence unless under control of strong promoter at high copy BBa_J04671 EYFP (RBS- LVA+ TERM): (E0032.B0015) intermediate building sequence BBa_E0434 EYFP (RBS+ AAV+ TERM): (B0034.E0034.B0015) BBa_E0036 EYFP (RBS- ASV+): more stable than E0032 or E0034 BBa_E0436 EYFP (RBS+ ASV+ TERM): (B0034.E0036.B0015) not listed as available BBa_J03102: YFP can be turned on by lambda and off by 434. Too bad it is stable and not ASV/AAV tagged… BBa_J06005: YFP, repressed by lambda and turned on by LuxR + HSL BBa_J06105: YFP, repressed by lambda and turned on by LasR + HSL BBa_S03476: lambda represses YFP. Has RBS + TERM BBa_S03475: lambda represses YFP. Has RBS + TERM. Has double lambda sites. BBa_S03466: lambda activates YFP. Has RBS + TERM
GREEN BBa_E0040 GFP BBa_J04031 GFP (LVA+): (E0040 and LVA modeled on E0032), listed as M BBa_J04631 GFP (RBS- LVA+ TERM): (J04031.B0015), intermediate building sequence BBa_J04431 GFP (LacI promo RBS+ LVA+ TERM): (R0010.B0034.J04031.B0015) BBa_E0240 GFP (RBS+ TERM): (B0032.E0040.B0015) medium rbs BBa_E0241 : like BBa_E0240 but has a different terminator – BRICK SHOWN TO WORK! BBa_E0840 GFP (RBS+ TERM): (B0030.E0040.B0015) strong rbs BBa_J07034 GFP (FecA’ promo, RBS+ TERM): (J07018.E0840), not listed as available BBa_J07011 GFP (ctx promo, RBS+ TERM): (J07007.E0840), not listed as available BBa_I7102 GFP (tetR promo, sRBS, TERM): (R0040.B0030.E0040.B0015), not listed as available BBa_I13522 GFP (tetR promo RBS+ TERM): (R0040.B0034.E0040.B0015) BBa_J04430 GFP (LacI promo RBS+ TERM): (R0010.B0034.E0040.B0015) BBa_E0044 GFP (AAV+) BBa_E0244 GFP (RBS+ AAV+ TERM): (B0032.E0044.B0015) medium rbs, not listed as available BBa_E0844 GFP (RBS+ AAV+ TERM): (B0030.E0044.B0015) strong rb, not listed as available BBa_S03336: lambda activates GFP. Has RBS + TERM BBa_S03335: lambda suppresses GFP. Has RBS + TERM
RED BBa_E1010 mRFP1 (July 2004) BBa_I13521 mRFP1 (tetR promo RBS+ TERM): (R0040.B0034.E1010.B0015) BBa_J04051 mRFP1 (LVA+): listed as M BBa_J04651 mRFP1 (RBS- LVA+ TERM): (J04051.B0015) intermediate building sequence BBa_J04451 mRFP1 (LacI promo RBS+ LVA+ TERM): (R0010.B0034.J04051.B0015), not listed as available BBa_J04450 mRFP1 (LacI promo RBS+ TERM): (R0010.B0034.E1010.B0015) BBa_J06504: mCherry BBa_J06505: mCherry + LVA BBa_J06702: mCherry + RBS + TERM BBa_J06703: mCherry + RBS + LVA + TERM BBa_S03473: lambda represses RFP. Has RBS + TERM. Has double lambda sites. BBa_S03337: lambda activates RFP. Has RBS + TERM
COMPOSITE BBa_I15011 ECFP+EYFP: (B0030.E0020.B0030.E0030) single mRNA containing ECFP and EYFP produced under strong RBS BBa_I15015 ECFP+EYFP: (B0032.E0020.B0032.E0030) medium RBS BBa_I15018 ECFP+EYFP: (B0031.E0020.B0031.E0030) weak RBS BBa_J13003 ECFP+EYFP: (E0020.B0034.E0030.B0010.B0012): CFP produced in presence of RIPS (specific ribosome?), YFP produced in its absence
LacZ BBa_E0033 lacZ alpha fragment (RBS-): portion of LacZ gene
BBa_E0666 : Lac I controls CFP, Tet R controls YFP, lambda controls mRFP. None are destabilized. This could be a useful oscillator. Can we get this made with destabilized colours? BBa_E0669: as above, but lambda turns on yellow, LacI controls cyan, TetR controls red. BBa_I13604: Tet R controls yellow, LacI controls cyan, no degradation tags. BBa_I13605: Lac I controls yellow, TetR controls cyan, no degradation tags BBa_I13607: Lac I controls yellow, TetR controls cyan, LVA degradation tags.
Signaling promoters with colour
BBa_J07066, RFP controlled by FecA. Needs FecB BBa_J13000: construct BBa_J13020 : LasR + A1 signal induces YFP BBa_J13021: LuxR + HSL makes YFP BBa_J13025: LuxR + HSL makes CFP BBa_J13026: LuxR + HSL makes RFP
Bba_M30000: LacZ controlled by osmosensor
SINGLE BLOCKS - PROMOTERS
BBa_I14018 P(Bla) , constitutive, medium BBa_I14033 P(Cat), constitutive medium BBa_I14034 P(Kat), constitutive medium, BBa_I14032 P(Lac) IQ, constitutive strong BBa_I0500 acrtivated by arabinose BBa_J03007 activated by maltose BBa_R0010 repressed by lac I BBa_R0011 activated by lambda, repressed by lac I BBa_R0051 repressed by lambda BBa_R1051 repressed by lambda BBa_I12007 Modified lambda Prm promoter (OR-3 obliterated), activated by lambda BBa_I12006 Modified lamdba Prm promoter (repressed by 434 cI, activated by lambda) BBa_I12036 Modified lamdba Prm promoter (cooperative repression by 434 cI, activation by lambda) – dual sites for better control BBa_I12010 Modified lamdba Prm promoter (repressed by p22 cII) BBa_R0053 reprressed by p22 BBa_R1053 reprressed by p22 BBa_R1052 repressed by 434 BBa_R0040 (tetR, negative) BBa_I1051 Lux cassette right promoter (needs HSL) BBa_R0061 Lux R + HSL represses BBa_R0062 Lux R + HSL activates BBa_R1062 Lux R + HSL activates BBa_R0063 (luxR & HSL regulated -- lux pL). Weak constitutive, shut down by Lux + HSL BBa_R0065 activated by Lux + HSL, repressed by lambda BBa_I14017 P(Rhl), needs HSL BBa_R0071 activated by RhlR + C4-HSL BBa_R0079 activated by LasR + HSL BBa_R0074 repressed by pen I
SINGLE BLOCKS - RIBOSOME BINDING SITES
BBa_B0030 RBS.1 (strong) BBa_B0031 RBS.2 (weak) BBa_B0032 RBS.3 (medium) BBa_B0033 RBS.4 (weakest) BBa_B0034 RBS (strongest)
SINGLE BLOCKS - TERMINATORS
BBa_B0010 Terminator Forward BBa_B0011 Terminator Bidirectional BBa_B0014 Terminator Forward BBa_B0015 Terminator Forward BBa_B0021 Terminator Bidirectional BBa_B0025 Terminator Reverse
SINGLE BLOCKS - ACTIVATORS, REPRESSORS
BBa_C0012 Repressor, lacI (LVA+) BBa_C0040 Repressor, tetR (LVA+) BBa_C0050 Repressor, HK022 cI (RBS- LVA+) BBa_C0051 Repressor, Lambda cI (RBS- LVA+) BBa_C0052 Repressor, 434 cI (RBS- LVA+) BBa_C0053 Repressor, P22 c2 (RBS- LVA+) BBa_C0062 Repressor/Activator, luxR BBa_C0071 Repressor/Activator, rhlR (LVA+) BBa_C0072 Repressor, mnt (strong) (LVA+) BBa_C0073 Repressor, mnt (weak) (LVA+) BBa_C0074 Repressor, penI (LVA+) BBa_C0171 repressor rhlR (-LVA) BBa_C0177 activator cinR (-LVA) BBa_C0077 Activator, cinR (LVA+) BBa_C0079 Activator, lasR (LVA+) BBa_C0179 represspor lasR (-LVA) BBa_C0080 Repressor/Activator, araC (LVA+) BBa_C0024 CheB, demethylates aspartate receptors (for chemotaxis) BBa_C0028 CheR coding region, methylates aspartate receptors (for chemotaxis)
SINGLE BLOCKS - ENZYMES
BBa_C0060 Enzyme, aiiA (LVA+), Makes signaling molecule BBa_C0160 Enzyme aiiA (-LVA) BBa_C0061 Enzyme, luxI (LVA+), Makes signaling molecule BBa_C0161 enyme luxI (-LVA) BBa_C0070 Enzyme, rhlI (LVA+), Makes signaling molecule BBa_C0170 enzyme rhlI (-LVA) BBa_C0078 Enzyme, lasI (LVA+), Makes signaling molecule BBa_C0178 Enzyme lasI (-LVA) BBa_C0076 Enzyme, cinI (LVA+), Makes signaling molecule BBa_C0176 enzyme cinI (-LVA) BBa_C0083 Enzyme, aspA (LVA+), makes aspartate BBa_C0020 CheY, makes bugs tumble
SINGLE BLOCKS - STICKY TAGS
BBa_M0020 GST tag binds glutathione BBa_M0021 Poly-His tag binds nickel, zinc, cobalt affinity resins BBa_M0024 biotin tag – binds streptavidin BBa_M0025 MBP tag – binds maltose BBa_M0027 CBP tag (cellulose binding peptide) BBa_M0030 Poly-Arg binds cation resin BBa_M0031 Strep-tag II binds biotin
SINGLE BLOCKS - TAGS
DEGRADATION TAGS
BBa_M0040 - LVA, slow degradation BBa_M0042 - LAA, medium degradation BBa_M0044 - AAV, fast degradation BBa_M0040 - ASV, fastest degradation
