Line 163: | Line 163: | ||
<details> | <details> | ||
<summary>BBa_K2500003: p<sub>TlpA</sub></summary> | <summary>BBa_K2500003: p<sub>TlpA</sub></summary> | ||
− | |||
<p><strong>Organism:</strong> <span class="bacterium">Salmonella typhimurium</span></p> | <p><strong>Organism:</strong> <span class="bacterium">Salmonella typhimurium</span></p> | ||
<p><strong>Original sequence:</strong> <a href="https://doi.org/10.1038/nchembio.2233">Piraner, Dan I., et al. <cite>Nature chemical biology</cite> 13.1 (2017): 75-80.</a></p> | <p><strong>Original sequence:</strong> <a href="https://doi.org/10.1038/nchembio.2233">Piraner, Dan I., et al. <cite>Nature chemical biology</cite> 13.1 (2017): 75-80.</a></p> | ||
− | <p><strong>Source:</strong> | + | <p><strong>Source:</strong> oligonucleotide sequence</p> |
<p><strong>Registry:</strong> <a href="http://parts.igem.org/Part:BBa_K2500003">BBa_K2500003</a></p> | <p><strong>Registry:</strong> <a href="http://parts.igem.org/Part:BBa_K2500003">BBa_K2500003</a></p> | ||
</details> | </details> | ||
Line 172: | Line 171: | ||
<details> | <details> | ||
<summary>BBa_K2500004: TlpA</summary> | <summary>BBa_K2500004: TlpA</summary> | ||
− | < | + | <p><strong>Organism:</strong> <span class="bacterium">Salmonella typhimurium</span></p> |
− | < | + | <p><strong>Original sequence:</strong> <a href="https://doi.org/10.1038/nchembio.2233">Piraner, Dan I., et al. <cite>Nature chemical biology</cite> 13.1 (2017): 75-80.</a></p> |
− | </ | + | <p><strong>Source:</strong> gBlock</p> |
− | <p> | + | <p><strong>Registry:</strong> <a href="http://parts.igem.org/Part:BBa_K2500004">BBa_K2500004</a></p> |
− | <p> Registry: <a href="http://parts.igem.org/Part:BBa_K2500004">BBa_K2500004</a></p> | + | |
</details> | </details> | ||
<details> | <details> | ||
<summary>BBa_K2500005: RBS_TlpA</summary> | <summary>BBa_K2500005: RBS_TlpA</summary> | ||
− | < | + | <p><strong>Organism:</strong> <span class="bacterium">Salmonella typhimurium</span></p> |
− | < | + | <p><strong>Original sequence:</strong> <a href="https://doi.org/10.1038/nchembio.2233">Piraner, Dan I., et al. <cite>Nature chemical biology</cite> 13.1 (2017): 75-80.</a></p> |
− | </ | + | <p><strong>Source:</strong> gBlock</p> |
− | <p> | + | <p><strong>Registry:</strong> <a href="http://parts.igem.org/Part:BBa_K2500005">BBa_K2500005</a></p> |
− | <p> Registry: <a href="http://parts.igem.org/Part:BBa_K2500005">BBa_K2500005</a></p> | + | |
</details> | </details> | ||
<details> | <details> | ||
<summary>BBa_K2500006: Protein E</summary> | <summary>BBa_K2500006: Protein E</summary> | ||
− | < | + | <p><strong>Organism:</strong> phage Phi X 174</p> |
− | < | + | <p><strong>Original sequence:</strong> provided by dr. Irene Wüthrich from Panke lab at the D-BSSE of ETH Zurich</p> |
− | </ | + | <p><strong>Source:</strong> plasmid provided by dr. Irene Wüthrich from Panke lab at the D-BSSE of ETH Zurich</p> |
− | <p> | + | <p><strong>Registry:</strong> <a href="http://parts.igem.org/Part:BBa_K2500006">BBa_K2500006</a></p> |
− | <p> Registry: <a href="http://parts.igem.org/Part:BBa_K2500006">BBa_K2500006</a></p> | + | |
</details> | </details> | ||
<details> | <details> | ||
<summary>BBa_K2500007: RBS<sub>eng</sub>_TlpA</summary> | <summary>BBa_K2500007: RBS<sub>eng</sub>_TlpA</summary> | ||
− | < | + | <p><strong>Design notes:</strong>FIXME FIXME FIXME</p> |
− | < | + | <p><strong>Organism:</strong> <span class="bacterium">Salmonella typhimurium</span></p> |
− | </ | + | <p><strong>Original sequence:</strong> <a href="https://doi.org/10.1038/nchembio.2233">Piraner, Dan I., et al. <cite>Nature chemical biology</cite> 13.1 (2017): 75-80.</a></p> |
− | <p> | + | <p><strong>Source:</strong> gBlock</p> |
− | <p> Registry: <a href="http://parts.igem.org/Part:BBa_K2500007">BBa_K2500007</a></p> | + | <p><strong>Registry:</strong> <a href="http://parts.igem.org/Part:BBa_K2500007">BBa_K2500007</a></p> |
</details> | </details> | ||
Line 209: | Line 205: | ||
<details> | <details> | ||
<summary>BBa_K2500008: p<sub>Const</sub>_RBS_LldP/LldR_p<sub>Const</sub>_RBS_LuxR</summary> | <summary>BBa_K2500008: p<sub>Const</sub>_RBS_LldP/LldR_p<sub>Const</sub>_RBS_LuxR</summary> | ||
− | < | + | <p><strong>Design notes:</strong>FIXME FIXME FIXME</p> |
− | < | + | <p><strong>Organism:</strong> <span class="bacterium">Salmonella typhimurium</span></p> |
− | </ | + | <p><strong>Original sequence:</strong> <a href="https://doi.org/10.1038/nchembio.2233">Piraner, Dan I., et al. <cite>Nature chemical biology</cite> 13.1 (2017): 75-80.</a></p> |
− | <p> | + | <p><strong>Source:</strong> gBlock</p> |
− | <p> Registry: <a href="http://parts.igem.org/Part: | + | <p><strong>Registry:</strong> <a href="http://parts.igem.org/Part:BBa_K2500007">BBa_K2500007</a></p> |
</details> | </details> | ||
Revision as of 09:30, 30 October 2017
![CATE logo](https://static.igem.org/mediawiki/2017/d/d6/T--ETH_Zurich--LogoMinimal.png)
Parts
![FIXME](https://static.igem.org/mediawiki/2017/7/7e/T--ETH_Zurich--basicparts.png)
Overview
We submitted in total 14 new BioBricks to the iGEM registry! You can find all parts and their design below.
Basic Parts
Part | Description | BioBrick |
---|---|---|
Bacterioferritin | Heme-deletion mutant of bacterial iron storage protein functioning as an MRI contrast agent | BBa_K2500000 |
Azurin | Redox protein originating from P. aeruginosa with anti-cancer activity | BBa_K2500001 |
p28 | Effector domain of azurin | BBa_K2500002 |
pTlpA | Temperature-responsive promoter optimized for slight activation above 37°C and full activation at 45 °C | BBa_K2500003 |
TlpA | Temperature-dependent transcriptional repressor of pTlpa | BBa_K2500004 |
RBS_TlpA | Temperature-dependent transcriptional repressor of pTlpa and synthetic RBS | BBa_K2500005 |
Protein E | Bacteria-lysing protein encoded by phage Phi X 17 | BBa_K2500006 |
RBSeng_TlpA | Temperature-dependent transcriptional repressor of pTlpa with engineered RBS | BBa_K2500007 |
RBSeng_ProteinE | Bacteria-lysing protein encoded by phage Phi X 174 with engineered RBS | BBa_K2500009 |
AND Gate A | Synthetic promoter responsive to LldR and luxR | BBa_K2500010 |
Best Basic Part: AND Gate B |
Synthetic promoter responsive to LldR and luxR | BBa_K2500011 |
AND Gate C | Synthetic promoter responsive to LldR and luxR | BBa_K2500012 |
pConst_RBS_LldP/ LldR_pConst_RBS_LuxR |
Expression cassette consisting of LuxR and LldP/LldR | BBa_K2500013 |
Composite Parts
Part | Description | BioBrick |
---|---|---|
Best Composite Part: pConstRBSeng_TlpA |
Constitutively expressed temperature-dependent transcriptional repressor of pTlpa with engineered RBS | BBa_K2500008 |
Design
All parts were codon-optimized for expression in E. coli Nissle via Geneious software and modified to remove forbidden restriction sites.
BBa_K2500000: Bacterioferritin
Organism: Escherichia coli Nissle
Original sequence: BBa_K1438001
Source: gBlock
Registry: BBa_K2500000
BBa_K2500001: Azurin
Design notes: Native signalling peptide removed.
Organism: Pseudomonas aeruginosa
Original sequence: BBa_K835004
Source: gBlock
Registry: BBa_K2500001
BBa_K2500002: p28
Design notes:Designed by taking aminoacids 50 to 77 from azurin and adding a start codon (ATG) at the beginning and two stop codons (TAA TAA) at the end.
Organism: Pseudomonas aeruginosa
Original sequence: BBa_K2500001
Source: gBlock
Registry: BBa_K2500002
BBa_K2500003: pTlpA
Organism: Salmonella typhimurium
Original sequence: Piraner, Dan I., et al. Nature chemical biology 13.1 (2017): 75-80.
Source: oligonucleotide sequence
Registry: BBa_K2500003
BBa_K2500004: TlpA
Organism: Salmonella typhimurium
Original sequence: Piraner, Dan I., et al. Nature chemical biology 13.1 (2017): 75-80.
Source: gBlock
Registry: BBa_K2500004
BBa_K2500005: RBS_TlpA
Organism: Salmonella typhimurium
Original sequence: Piraner, Dan I., et al. Nature chemical biology 13.1 (2017): 75-80.
Source: gBlock
Registry: BBa_K2500005
BBa_K2500006: Protein E
Organism: phage Phi X 174
Original sequence: provided by dr. Irene Wüthrich from Panke lab at the D-BSSE of ETH Zurich
Source: plasmid provided by dr. Irene Wüthrich from Panke lab at the D-BSSE of ETH Zurich
Registry: BBa_K2500006
BBa_K2500007: RBSeng_TlpA
Design notes:FIXME FIXME FIXME
Organism: Salmonella typhimurium
Original sequence: Piraner, Dan I., et al. Nature chemical biology 13.1 (2017): 75-80.
Source: gBlock
Registry: BBa_K2500007
BBa_K2500008: pConst_RBS_LldP/LldR_pConst_RBS_LuxR
Design notes:FIXME FIXME FIXME
Organism: Salmonella typhimurium
Original sequence: Piraner, Dan I., et al. Nature chemical biology 13.1 (2017): 75-80.
Source: gBlock
Registry: BBa_K2500007
BBa_K2500010: AND gate A
![FIXME](https://static.igem.org/mediawiki/2017/7/72/T--ETH_Zurich--ANDgateA.png)
In the absence of high concentrations of L-lactate, LldR inhibitor proteins bind to the binding sites O1 and O2 surrounding the pLux promoter leading to the formation of a DNA loop. The pLux promoter is sequestered and inaccessible for transcription. In design A, the distances between the intercalated promoter and the binding sites were adapted from BBa_K1847007.
Registry: BBa_K2500010
BBa_K2500011: AND Gate B
![FIXME](https://static.igem.org/mediawiki/2017/6/6a/T--ETH_Zurich--ANDgateB.png)
In design B, each binding site was duplicated in order to achieve a potential zipper mechanism and stronger inhibition due to binding more LldR inhibitor proteins.
Registry: BBa_K2500011
BBa_K2500012: AND Gate C
![FIXME](https://static.igem.org/mediawiki/2017/c/c2/T--ETH_Zurich--ANDgateC.png)
In design C, an artificial spacer was embedded between the pLux promoter and the O2 binding site in order to influence the looping dynamics.
Registry: BBa_K2500012