Team:INSA-UPS France/Parts
Feel free to contact us, if you want to know more about our parts: igem.toulouse@gmail.com
BBa_J04450 was tested in the Vibrio harveyi background. The biobrick was cloned in a broad host range plasmid (pBBR1MCS-4) and conjugated into Vibrio harveyi to demonstrate the production of RFP in this chassis.
To learn more: see <partinfo>BBa_J04450</partinfo> and our results.
In this part encoding sequence of D-NY15 was placed under the yeast constitutive promoter pGAP. This part was characterized by RT-qPCR.
To learn more: see <partinfo>BBa_K431009</partinfo> and our results.
The α-factor sequence contains a kozak region and a signal sequence to secrete the produced peptides. The functionality of the signal factor was investigated by demonstrating that AMP are present in the supernatant using a toxicity assay.
To learn more: see <partinfo>BBa_K180001</partinfo> and our results.
This part includes Odr-10 receptor under the control of pGAP, a constitutive yeast promoter. When diacetyl binds to Odr-10 a cascade of activation of Ste proteins (endogenous to P. pastoris ) will lead to the binding of Ste12 on pFUS1 promoter, and the expression of RFP should be activated.
To learn more: see <partinfo>BBa_K1072010</partinfo>, <partinfo>BBa_K1072023</partinfo> and our results.
This part contains the first half of Vibrio harveyi quorum CqsS receptor which is able to detect CAI-1 family molecule. The receptor is under the control of a constitutive promoter.
This part has been designed due to IDT gBlocks synthesis requirements. The ordered gBlocks size should not exceed 3kb (the total length of the Vibrio harveyi circuit is 6929 bp). The XhoI restriction site naturally present in the gene was used reassemble the cqsS* sequence.
Therefore, it was impossible to use pSB1C3 vector because of the presence of a XhoI restriction on the vector. This part was then subcloned in pBR322.
To learn more: see our design, and our results.
This part contains the second half of Vibrio harveyi quorum CqsS* receptor which is able to detect CAI-1 family molecule. It also contains the tetR repressor under the control of pQRR4 promoter.
To learn more: see our design, and our results.
This part includes the complete cqsS* gene under a constitutive promoter and the tetracyclin repressor under the control of pQRR4 promoter which activation depends on the detection of quorum sensing inducers by CqsS*.
To learn more: see our design, and our results.
This part includes the complete cqsS under the control of a constitutive promoter and the tetracycline repressor (TetR) under the control of pQRR4 promoter which activation depends on CqsS* recognition of quorum sensing inducers. The als gene, implied in the diacetyl production pathway, is under the control of pTet which can be repressed by TetR.
To learn more: see our design, and our results.
This system allowed us to constitutively express BBa_K2278021 with a yeast promoter. This system was used to produce AMP at the high concentration.
To learn more: see <partinfo>BBa_K2278021</partinfo>, and our results.
This system allowed us to constitutively express the BBa_K2278022 with a yeast promoter. This system was used to produce AMP at high concentration.
To learn more: see <partinfo>BBa_K2278022</partinfo> and see our results.
This system allowed us to constitutively express BBa_K2278023 with a yeast promoter. This system was used to produce AMP at high concentration.
To learn more: see <partinfo>BBa_K2278023</partinfo> and our results.
This system allowed us to constitutively express Odr10 receptor to detect diacetyl at any time. Odr10 pathway leads to the activation of pFUS promoter when diacetyl is present in the environment, hence the AMP are produced.
To learn more: see <partinfo>BBa_K2278023</partinfo>, and our results.
iGEM UPS-INSA Toulouse 2017
Parts
*** All parts listed here are available upon request ***
New parts submitted to the registry
Name
Function
Type
Part
State
<partinfo>BBa_K2278001</partinfo>
QS molecule generator
basic
working
<partinfo>BBa_K2278002</partinfo>
QS molecule generator
basic
not tested
<partinfo>BBa_K2278011</partinfo>
Diacetyl generator
basic
issues
<partinfo>BBa_K2278021</partinfo>
D-NY15 AMP generator
basic
working
<partinfo>BBa_K2278022</partinfo>
Leucrocin I AMP generator
basic
unsuccessful
<partinfo>BBa_K2278023</partinfo>
coT2 AMP generator
basic
unsuccessful
Existing Parts we have contributed to characterized
<partinfo>BBa_J04450</partinfo>: RFP coding device
<partinfo>BBa_K431009</partinfo>: glyceraldehyde 3-phosphate dehydrogenase promoter (pGAP)
<partinfo>BBa_K1800001</partinfo>: alpha factor secretion signal
Pichia pastoris complete module with reporter gene : Odr-10 diacetyl receptor (<partinfo>BBa_K1072010</partinfo>), pFUS1 (<partinfo>BBa_K1072023</partinfo>)
This system allowed to produce RFP in response to diacetyl.
This construction was cloned in pPICZalpha yeast vector.
Parts used in our project but not submitted to the registry
Vh1: First part of Vibrio harveyi gene circuit
Vh 2 : Second part of Vibrio harveyi gene circuit
This part has been designed due to IDT gBlocks synthesis requirements. The ordered gBlocks size should not exceed 3kb (the total length of the Vibrio harveyi circuit is 6929 bp). The ApaI (not present on pSB1C3 vector) and XhoI restriction (naturally present in the cqsS* gene) were used to reassemble the synthetic circuit.
Vh 1+2: Vibrio harveyi invertor system
Vh1+2 is the assembly of Vh2 in Vh1. This part was cloned in pBR322.
Vibrio harveyi complete gene circuit
This part is the assemblage of Vh1+2 in BBa_K2278011. It allows to produce diacetyl is produced upon detection of Vibrios CAI-1 quorum sensing molecule.
The als gene is flanked with two restriction enzyme sites to clone a reporter gene such as rfp.
The assembly of this part was not released due to technical and time issues.
Pichia pastoris D-NY15 AMP generator
The α-factor sequence contains a RBS and a signal sequence to secrete the produced peptides.
The restriction enzyme sites allow to extract individually each components of the plasmid. This construction was cloned in pPICZalpha yeast vector.
Pichia pastoris Leucrocin I AMP generator
The α-factor sequence contains a RBS and a signal sequence to secrete the produced petides.
The restriction enzyme sites allow to extract individually each components of the plasmid. This construction was cloned in pPICZalpha yeast vector.
Pichia pastoris cOT2 AMP generator
The α-factor sequence contains a RBS and a signal sequence to secrete the produced peptides.
The restriction enzyme sites allow to extract individually each components of the plasmid.
This construction was cloned in pPICZalpha yeast vector.
Pichia pastoris complete module
The α-factor signal and coT2 gene are flanked with two restriction enzyme sites to clone a reporter gene such as RFP.
This construction was cloned in pPICZalpha yeast vector.
Strategy pages
Description
Design
Parts
