Team:Stanford/PartsPage

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<th> Cloning Step </th><th> Basic Parts </th><th> Detailed Parts </th><th> Picture </th>
<th> Cloning Step </th><th> Basic Parts </th><th> Detailed Parts </th><th> Picture </th>

Revision as of 00:25, 22 October 2009

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Contents

Stanford iGEM 2009 Registered Parts

You can find the Stanford iGEM registered parts for 2009 as well as the team's favorite parts at:

Stanford in the Parts Registry

Table of Registered Parts

Part #NamePart DescriptionType
BBa_K223040 SoxR GeneThis part produces the SoxR protein, which in the presence of oxidative stresses, such as paraquat and nitric oxide, binds with high affinity to the SoxS promoter. The SoxR generator, when coupled with the SoxS promoter, can be utilized as an inducible sensor for oxidative stress.Basic
Bba_K223041SoxS promoterThe SoxS promoter is activated by the SoxR protein in the presence of oxidative stress, such as Paraquat or Nitric Oxide. It has a high affinity binding with the SoxR protein. It can be used in conjunction with the SoxR gene as an inducible sensor for oxidative stress. Basic
Bba_K223042Blh GeneThe Blh gene cleaves beta-carotene into retinal. It can be used in the pathway for the production of retinoic acid from beta-carotene in conjunction with the RALDH II genes. Basic
Bba_K223043RALDH II GeneThe Raldh II gene converts retinal into retinoic acid. It is helpful if used in conjunction with the Blh gene for the entire process of converting beta-carotene into retinoic acid.Basic
Bba_K223044SoxR-SoxS SystemThis entire system works as a superoxide sensor. Superoxides activate the SoxR protein to bind with the SoxS Promoter and begin transcription of downstream genes. It is an inducible sensor.Composite
Bba_K223045 RALDH II IntermediateWorking composite part with a ribosome binding site and terminator.Intermediate
Bba_K223046SoxR/SoxS- GFPSoxR - SoxS Promoter System with downstream GFP production. This was used to characterize the inducible oxidative stress signal when activating the SoxR-SoxS promoter system.Composite
Bba_K223047Lacl Promoter-SoxR/SoxS-GFPThis is the SoxR-SoxS inducible promoter system with downstream production of GFP in order to characterize the device. We used the lacl regulated promoter.Composite
Bba_K223048Blh and RALDH IIThis is the RBS- Blh- RBS- RALDH II- TT generator. Both of these parts are important in the cleaving of beta carotene into retinoic acid.Intermediate
Bba_K223049SoxR - SoxS- Blh and RALDH IIThis is the superoxide inducible SoxR-SoxS promoter system coupled with the production of Blh and RALDH II genes for the cleaving of beta carotene into retinoic acid.Composite
Bba_K223050Lacl Promoter-SoxR - SoxS -Blh and RALDH II Same as BBa_K223049 with the addition of an inducible lacl promoter.Composite
Bba_K223051 5MT operon (promoter-operator)This is a modified trp operon, promoter and operator site, that recognizes and binds to a doubly mutant repressor and its corepressor, 5-methyl-L-tryptophan. In conjunction with the mutant tryptophan repressor, this modified operon functions as an inverter, converting decreasing levels of 5MT into a PoPs output signal for downstream transcription.Basic
Bba_K2230525MT Mutant TrpR geneThe 5 methyl-L-tryptophan aporepressor gene is a mutant version of the regular TrpR gene involved in sensing levels of synthetic compound, 5 MT. It works by binding to its corepressor, 5 MT, and then to its corresponding promoter operator, BBa_K223051, repressing downstream transcription. In the presence of low corepressor levels, the 5MT aporepressor will not bind allowing for downstream transcription.Basic
Bba_K223053hIL-6 Generator (Freiburg-compatible)This gene generates Interleukin 6, an important cell signaling molecule that acts as both a pro- and anti-inflammatory cytokineBasic
Bba_K223054HlyA Signal Sequence (Freiburg-compatible)The Hemolysin A signal sequence is generally fused with a second protein. Together with the Hly complex (HlyR, HlyC, HlyB and HlyD) and TolC, HlyAs can be used to take large proteins out of the cell. Basic
Bba_K223055TolC GeneTolC, along with HemolysinB, C, and D, for a channel in the cellular membrane to allow for the extracellular transport of HlyAs-fused protein sequences. The TolC receptor, in conjunction to the Hly genes, can serve as channel for transporting large proteins.Basic
Bba_K223056Front Flanking Sequence (HlyR, IS1, HlyC)This front flanking sequence has three of the hemolysin parts - HlyR, Is1 and HlyC. In conjunction with rear flanking sequence, containing HlyB and HlyD, and TolC (part K223055), this part creates a channel protein to facilitate the extracellular export HlyAs-fused proteins.Basic
Bba_K223057 Rear Flanking Sequence (HlyB and HlyD)Along with parts BBa_K223055 and K223056, this part helps form the channel protein for the extracellular transport of HlyAs-fused protein.Basic
Bba_K223058RBS+mutTrpR+TT+mutTrp OperatorThis part is a combination of K223051 and K223052. It is an inverter that is sensitive to concentration decreases in 5-methyl-L-tryptophan and converts this to a PoPs output signal used for downstream transcription.Composite
Bba_K223059mutTrp (5MT) Inverter with GFP Biobrick partA modification to Biobrick part BBa_K223058. It has a GFP biobrick part, BBa_I13504, at the end of the sequence and functions as a reporter.Composite
Bba_K2230605MT Inverter-GFP Reporter and Lac PromoterIdentical to part BBa_K223059 except with a Lac promoter. This part can be used to test the efficiency of the 5MT inverter system.Composite
Bba_K223061hIL-6 + HlyAs Fusion protein (Freiburg Compatible) This sequence when coupled with TolC (BBa_K223055), Front Flanking (BBa_K223056) and Rear Flanking (BBa_K223057) sequences, allows for the extracellular export of hIL-6. This particular part only forms the signal sequence for export.Composite
Bba_K223062TolC and Hemolysin B, C and D gene complexThis a composite part of BBa_K223055, BBa_K223056, BBa_K223057 that provides all necessary genes to create a channel protein in the extra cellular membrane of a prokaryotic cell. This channel can be used to export large organic molecules fused to a signal sequence, HlyAsComposite
Bba_K223063RBS+BLHIdentical to BBa_K223042 but with RBS (B0034) prior to the sequence. Composite

Device 1 Cloning Plan with Proper Sequences

Cloning Step Basic Parts Detailed Parts Picture
Step 1.1a SoxR/S Promoter + GFP tag RBS-SoxR-TT-SoxS Promoter + RBS-GFP-TT 1.1a
Step 1.1b SoxR/S Promoter + Blh RBS-SoxR-TT-SoxS Promoter + RBS-Blh 1.1b
Step 1.1c Blh + Terminator RBS-Blh + TT 1.1c
Step 1.1d Constitutive Promoter + Blh (1.1c) Promoter +RBS-Blh-TT 1.1d
Step 1.1e Constitutive Promoter + Crt Cluster Promoter + RBS-Crt 1.1e
Step 1.2a Constitutive Promoter + SoxR/s w/ GFP (1.1a) Promoter+ RBS-SoxR-TT-SoxSP-RBS-GFP-TT 1.2a
Step 1.2b SoxR/S-Blh (1.1b) + RALDH II RBS-SoxR-TT-SoxSP-RBS-Blh + RBS-RALDH II-TT 1.2b
Step 1.3a Constitutive Promoter + SoxR/S w/ Blh and RaldhII Promoter+ RBS-SoxR-TT-SoxSP-RBS-Blh-RBS-RALDH II-TT 1.3a
Controls W/ PQ (+) Constitutive Promoter + GFP tag Promoter+ RBS-GFP-TT 1.3a
Controls W/ PQ (-) GFP tag w/ No Promoter RBS-GFP-TT 1.3a
Controls Blh and Raldh II (+) Constitutive Promoter + Blh and RaldhII Promoter+ RBS-Blh-TT-RBS-RALDH II-TT 1.3a
Controls Blh and Raldh II (-) Blh and RaldhII w/ no Promoter RBS-Blh-TT-RBS-RALDH II-TT 1.3a

Device 2 Cloning Plan with Proper Sequences

Cloning Step Basic Parts Detailed Parts Picture
Step 2.1a hIl-6-HlyAs + Terminator hIl-6-HlyAs + TT 1.1a
Step 2.1b hIl-6-HlyAs + YFP hIl-6-HlyAs + YFP 1.1b
Step 2.1c RBS + TolC RBS+ TolC 1.1c
Step 2.1d Terminator + HlyR TT + HlyR 1.1d
Step 2.1e Forward Flanking Sequence (FFS) + RBS Hly-C + RBS 1.1e
Step 2.1f Rear Flanking Sequence (RFS) + Terminator HlyB,D + TT 1.1e
Step 2.2a RBS + hIL-6-HlyAs (2.1a) RBS + hIL-6-HlyAs-TT 1.2a
Step 2.2b hIl-6-HlyAs-YFP(2.1b) + Terminator hIl-6-HlyAs-YFP+ TT 1.2b
Step 2.2c TolC (2.1c) + HlyR Promoter Element (2.1d) RBS-TolC + TT-HlyR 1.2b
Step 2.2d FFS-RBS (2.1e) + RFS-TT (2.1f) FFS-RBS + RFS-TT 1.2b
Step 2.3a Trp Operator + hIL-6-HlyAs (2.2a) Trp Op + RBS-hIL-6-HlyAs-TT 1.3a
Step 2.3b RBS + hIL-6-HlyAs-YFP-TT (2.2b) RBS + hIL-6-HlyAs-YFP-TT 1.3a
Step 2.3c TolC-HlyR (2.2c) + FFS-RFS (2.2d) RBS-TolC-TT-HlyR + FFS-RBS-RFS-TT 1.3a
Step 2.4a Trp Operator + HIl-6-HlyAs-YFP (2.3b) Trp Op + RBS-HIl-6-HlyAs-YFP-TT 1.3a
Step 2.4b Const. Promoter + TolC-HlyR-FFS-RFS (2.3c) Const. Promoter + RBS-TolC-TT-HlyR-FFS-RBS-RFS-TT 1.3a