Overview
This year, BASIS-China contributed a total of 12 basic and 9 composite parts. As for the composite parts, we’ve designed complete pathways including operator, RBS, and signal peptide for the sake of future iGEMers to use directly. In parallel, we borrowed a suicidal system from last year’s contribution to make our product safer by limiting the risk of manufactured organisms affecting nature.
Basic Part
This year, team BASIS-China planned to degrade microplastics with PETase. We’ve tested the distribution kit and decided to use AI to predict the mutation site of IsPETase and BhrPETase to increase their enzymatic efficiencies. Besides, we’ve also read multiple latest academic essays and used some of the existing PETase that were proved to be efficient.
| Registry Code | Part Name | Short Description | Status | Base Pair |
| BBa_K5236002 | IsPETase M10L | This basic part encodes for a mutated IsPETase
M10L. | Succeed | 891 |
| BBa_K5236003 | IsPETase K95S | This basic part encodes for a mutated IsPETase
K95S. | Reconstructing sequence | 891 |
| BBa_K5236004 | IsPETase H104W | This basic part encodes for a mutated IsPETase
H104W. | Succeed | 891 |
| BBa_K5236005 | IsPETase M128L | This basic part encodes for a mutated IsPETase
M128L. | Repeat electrophoresis | 891 |
| BBa_K5236006 | IsPETase M154L | This basic part encodes for a mutated IsPETase M154L and is derived from Escherichia coli. | Succeed | 891 |
| BBa_K5236007 | IsPETase W159HF229Y | This basic part encodes for a IsPETase W159HF229Y and is derived from Escherichia coli. | Succeed | 891 |
| BBa_K5236008 | IsPETase S121ER224QN233K | This basic part encodes for a IsPETase S121ER224QN233K and is derived from Escherichia coli. | Succeed | 891 |
| BBa_K5236009 | BhrPETase W229F | This basic part encodes for a mutated BhrPETase W229F and is derived from Escherichia coli. | Succeed | 798 |
| BBa_K5236010 | BhrPETase N205G | This basic part encodes for a mutated BhrPETase N205G and is derived from Escherichia coli. | Succeed | 798 |
| BBa_K5236011 | BhrPETase N191S | This basic part encodes for a mutated BhrPETase N191S and is derived from Escherichia coli. | Succeed | 798 |
| BBa_K5236012 | BhrPETase M57L | This basic part encodes for a mutated BhrPETase M57L and is derived from Escherichia coli. | Reconstructing sequence | 798 |
| BBa_K5236013 | LCCICCG | This basic part encodes for a wild-type LCCICCG and is derived from Escherichia coli. | Succeed | 798 |
| BBa_K5236025 | BhrPETase | This basic part encodes for a wild-type BhrPETase and is derived from Escherichia coli. in our lab. | Succeed | 798 |
Composite Part
This year, team BASIS-China planned to degrade microplastics with PETase. We’ve tested the distribution kit and decided to use AI to predict the mutation site of IsPETase and BhrPETase to increase their enzymatic efficiencies. Besides, we’ve also read multiple latest academic essays and used some of the existing PETase that were proved to be efficient.
| Registry Code | Part Name | Short Description | Status | Base Pair |
| BBa_K5236014 | pET28a-pelB-IsPETase(M10L)-6xHis | T7 promoter: used to regulate gene expression of recombinant proteins.
Lac operator: used to control gene expression by lactose.
RBS: the location to bind ribosome.
IsPETase M10L: This sequence encodes for a mutated IsPETase M10L and is derived from Escherichia coli.
pelB: the sequence codes for signal peptide which helps PETase exit the cell membrane.
6xHis: a tag for the purification of protein which focus in the production of recombinant proteins.
T7 terminator: terminates transcription. | Circuit works | 1167 |
| BBa_K5236015 | pET28a-pelB-IsPETase(K95S)-6xHis | T7 promoter: used to regulate gene expression of recombinant proteins.
Lac operator: used to control gene expression by lactose.
RBS: the location to bind ribosome.
IsPETase K95S: This sequence encodes for a mutated IsPETase K95S and is derived from Escherichia coli.
pelB: the sequence codes for signal peptide which helps PETase exit the cell membrane.
6xHis: a tag for the purification of protein which focus in the production of recombinant proteins.
T7 terminator: terminates transcription. | Reconstructing sequence | 1167 |
| BBa_K5236016 | pET28a-pelB-IsPETase(H104W)-6xHis | T7 promoter: used to regulate gene expression of recombinant proteins.
Lac operator: used to control gene expression by lactose.
RBS: the location to bind ribosome.
IsPETase H104W: This sequence encodes for a mutated IsPETase H104W and is derived from Escherichia coli.
pelB: the sequence codes for signal peptide which helps PETase exit the cell membrane.
6xHis: a tag for the purification of protein which focus in the production of recombinant proteins.
T7 terminator: terminates transcription. | Circuit works | 1167 |
| BBa_K5236017 | pET28a-pelB-IsPETase(M128L)-6xHis | T7 promoter: used to regulate gene expression of recombinant proteins.
Lac operator: used to control gene expression by lactose.
RBS: the location to bind ribosome.
IsPETase M128L: This sequence encodes for a mutated IsPETase M128L and is derived from Escherichia coli.
pelB: the sequence codes for signal peptide which helps PETase exit the cell membrane.
6xHis: a tag for the purification of protein which focus in the production of recombinant proteins.
T7 terminator: terminates transcription. | Repeat electrophoresis | 1167 |
| BBa_K5236018 | pET28a-pelB-IsPETase(M154L)-6xHis | T7 promoter: used to regulate gene expression of recombinant proteins.
Lac operator: used to control gene expression by lactose.
RBS: the location to bind ribosome.
IsPETase M154L: This sequence encodes for a mutated IsPETase M154L and is derived from Escherichia coli.
pelB: the sequence codes for signal peptide which helps PETase exit the cell membrane.
6xHis: a tag for the purification of protein which focus in the production of recombinant proteins.
T7 terminator: terminates transcription. | Circuit works | 1167 |
| BBa_K5236019 | pET28a-pelB-BhrPETase(W229F)-6xHis | T7 promoter: used to regulate gene expression of recombinant proteins.
Lac operator: used to control gene expression by lactose.
RBS: the location to bind ribosome.
BhrPETase W229F: This sequence encodes for a mutated BhrPETase W229F and is derived from Escherichia coli.
pelB: the sequence codes for signal peptide which helps PETase exit the cell membrane.
6xHis: a tag for the purification of protein which focus in the production of recombinant proteins.
T7 terminator: terminates transcription. | Circuit works | 1251 |
| BBa_K5236020 | pET28a-pelB-BhrPETase(N205G)-6xHis | T7 promoter: used to regulate gene expression of recombinant proteins.
Lac operator: used to control gene expression by lactose.
RBS: the location to bind ribosome.
BhrPETase N205G: This sequence encodes for a mutated BhrPETase N205G and is derived from Escherichia coli.
pelB: the sequence codes for signal peptide which helps PETase exit the cell membrane.
6xHis: a tag for the purification of protein which focus in the production of recombinant proteins.
T7 terminator: terminates transcription. | Circuit works | 1251 |
| BBa_K5236021 | pET28a-pelB-BhrPETase(N191S)-6xHis | T7 promoter: used to regulate gene expression of recombinant proteins.
Lac operator: used to control gene expression by lactose.
RBS: the location to bind ribosome.
BhrPETase N191S: This sequence encodes for a mutated BhrPETase N191S and is derived from Escherichia coli.
pelB: the sequence codes for signal peptide which helps PETase exit the cell membrane.
6xHis: a tag for the purification of protein which focus in the production of recombinant proteins.
T7 terminator: terminates transcription. | Circuit works | 1251 |
| BBa_K5236022 | pET28a-pelB-BhrPETase(M57L)-6xHis | T7 promoter: used to regulate gene expression of recombinant proteins.
Lac operator: used to control gene expression by lactose.
RBS: the location to bind ribosome.
BhrPETase M57L: This sequence encodes for a mutated BhrPETase M57L and is derived from Escherichia coli.
pelB: the sequence codes for signal peptide which helps PETase exit the cell membrane.
6xHis: a tag for the purification of protein which focus in the production of recombinant proteins.
T7 terminator: terminates transcription. | Reconstructing sequence | 1251 |
Biocontainment system
Since a type II antitoxin-toxin (TA) system is functioning effectively as a suicidal system for killing our manufactured leakage microorganisms, we were inspired by last year’s contribution and build the exact same sequence pathway using the parts listed below.
| Registry Code | Part Name | Short Description | Status | Base Pair |
| BBa_K4880022 | PpsbA2-sepA1-PisiA7942-sepT1 | PempK-like type II TA system where the toxin is controled by PisiA7942 | Circuit works | 1272 |
| BBa_K4880023 | PpsbA2-sepA2-PisiA7942-sepT2 | VapC-like type II TA system where the toxin is controled by PisiA7942 | Circuit works | 1356 |
| BBa_K4880008 | sepA1 | This gene encodes for the antitoxin sepA1 and is derived from the large plasmid pANL of Synechococcus 7942. | Works | 216 |
| BBa_K4880009 | sepT1 | This gene encodes for the toxin sepT1 and is derived from the large plasmid pANL of Synechococcus 7942. | Works | 351 |
| BBa_K4880010 | sepA2 | This gene encodes for the antitoxin sepA2 and is derived from the large plasmid pANL of Synechococcus 7942. | Works | 267 |
| BBa_K4880011 | sepT2 | This gene encodes for the toxin sepT2 and is derived from the large plasmid pANL of Synechococcus 7942. | Works | 384 |
| BBa_K4880013 | PisiA7942 | This sequence is a Fe3+ repressed promoter derived from Synechococcus elongatus PCC 7942. | Works | 270 |