Composite Parts
Our composite parts are PhytoBricks for the organism Chlamydomonas reinhardtii which have been assembled with the Modular Cloning System, a way of Golden Gate cloning. All composite parts are listed below.
| Name | Type | Description | Designer |
| BBa_K3002100 | Composite | Level 1 - Mutant PETase + cCA + SP20 HA (Phytobrick) | Dorothée Klein |
| BBa_K3002101 | Composite | Level 1 – Mutant PETase + HA (Phytobrick) | Dorothée Klein |
| BBa_K3002102 | Composite | Level 1 - Spectinomycin Resistance Gene for Chlamydomonas reinhardtii (Phytobrick) | Dorothée Klein |
| BBa_K3002103 | Composite | Level 1 – Wildtype PETase + His (Phytobrick) | Dorothée Klein |
| BBa_K3002104 | Composite | Level 1 – Wildtype PETase + cCA + HA (Phytobrick) | Dorothée Klein |
| BBa_K3002105 | Composite | Level 1 – Mutant PETase + HA (Phytobrick) | Dorothée Klein |
| BBa_K3002106 | Composite | Level 1 – Mutant PETase + cCA + HA (Phytobrick) | Dorothée Klein |
| BBa_K3002107 | Composite | Level 1 – Mutant PETase + GLE + HA (Phytobrick) | Dorothée Klein |
| BBa_K3002108 | Composite | Level 1 - Mutant PETase + ARS + HA (Phytobrick) | Dorothée Klein |
| BBa_K3002109 | Composite | Level 1 - Mutant PETase + cCA + SP20HA (Phytobrick) | Dorothée Klein |
| BBa_K3002110 | Composite | Level 1 - Mutant PETase + GLE + SP20HA (Phytobrick) | Dorothée Klein |
| BBa_K3002111 | Composite | Level 1 – Mutant PETase + ARS + sp20-HA | Marlene Schlosser |
| BBa_K3002112 | Composite | Level 1 – MHETase + HA | Marlene Schlosser |
| BBa_K3002113 | Composite | Level 1 – MHETase + cCA + HA | Marlene Schlosser |
| BBa_K3002114 | Composite | Level 1 – MHETase + cCA + sp20-HA | Marlene Schlosser |
| BBa_K3002119 | Composite | Level 1 – Hygromycin resistance + Tub2 Promoter + Tub2 Terminator | Marlene Schlosser |
| BBa_K3002120 | Composite | Level 1 – Mutant PETase + cCA + sp20-His | Marlene Schlosser |
| BBa_K3002121 | Composite | Level 1 – Mutant PETase + ARS + sp20-His | Marlene Schlosser |
| BBa_K3002122 | Composite | Level 1 – Mutant PETase + GLE + sp20-His | Marlene Schlosser |
| BBa_K3002123 | Composite | Level 1 – MHETase + cCA + sp20-His | Marlene Schlosser |
| BBa_K3002124 | Composite | Level 1 – Wildtype-PETASE + cCA + sp20-His | Marlene Schlosser |
| BBa_K3002125 | Composite | Level 1 – Wildtype-PETase + ARS + sp20-His | Marlene Schlosser |
| BBa_K3002126 | Composite | Level 1 – Wildtype-PETase + His | Marlene Schlosser |
| BBa_K3002127 | Composite | Level 1 – Hygromycin Resistance Gene for Chlamydomonas reinhardtii (Phytobrick) | Dorothée Klein |
| BBa_K3002200 | Composite | L2 spectinomycin resistance + Mut-PETase + MHETase | Dorothée Klein |
| BBa_K3002202 | Composite | L2 spectinomycin resistance + cCA_Mut-PETase + cCA_MHETase | Dorothée Klein |
| BBa_K3002207 | Composite | L2 spectinomycin resistance + GLE_Mut-PETase | Dorothée Klein |
| BBa_K3002208 | Composite | L2 spectinomycin resistance + ARS_Mut-PETase | Dorothée Klein |
| BBa_K3002209 | Composite | L2 spectinomycin resistance + cCA_Mut-PETase | Dorothée Klein |
| BBa_K3002210 | Composite | L2 spectinomycin resistance + ARS_Mut-PETase + cCA_MHETase | Dorothée Klein |
| BBa_K3002211 | Composite | L2 spectinomycin resistance + GLE_Mut-PETase + cCA_MHETase | Dorothée Klein |
| BBa_K3002212 | Composite | L2 spectinomycin resistance + cCA_Mut-PETase_SP20HA + cCA_MHETase_SP20HA | Dorothée Klein |
| BBa_K3002213 | Composite | L2 spectinomycin resistance + ARS_Mut-PETase_SP20HA + cCA_MHETase_SP20HA | Dorothée Klein |
| BBa_K3002214 | Composite | L2 spectinomycin resistance + GLE_Mut-PETase_SP20HA + cCA_MHETase_SP20HA | Dorothée Klein |
| BBa_K3002215 | Composite | L2 spectinomycin resistance + L1_connector_Pos2 + ccA_MHETase_SP20HA | Dorothée Klein |
| BBa_K3002216 | Composite | L2 spectinomycin resistance + L1_connector_Pos2 + ccA_MHETase | Dorothée Klein |
| BBa_K3002217 | Composite | L2 spectinomycin resistance + cCA_Mut-PETase_SP20HA | Dorothée Klein |
| BBa_K3002218 | Composite | L2 Hygromycin resistance + L1_connector_Pos2 + ccA_MHETase_SP20HA | Dorothée Klein |
| BBa_K3002219 | Composite | L2 spectinomycin resistance + ARS_Mut-PETase_SP20HA | Dorothée Klein |
| BBa_K3002220 | Composite | L2 spectinomycin resistance + GLE_Mut-PETase_SP20HA | Marlene Schlosser |
| BBa_K3002221 | Composite | L2 spectinomycin resistance + cCA_Mut-PETase_SP20His | Marlene Schlosser |
| BBa_K3002222 | Composite | L2 spectinomycin resistance + ARS_Mut-PETase_SP20His | Marlene Schlosser |
| BBa_K3002223 | Composite | L2 spectinomycin resistance + GLE_Mut-PETase_SP20His | Marlene Schlosser |
| BBa_K3002224 | Composite | L2 spectinomycin resistance + cCA_Mut-PETase_SP20-His + ccA_MHETase_HA-SP20HA | Marlene Schlosser |
| BBa_K3002225 | Composite | L2 spectinomycin resistance + GLE_Mut-PETase_SP20His + ccA_MHETase_SP20HA | Marlene Schlosser |
| BBa_K3002226 | Composite | L2 spectinomycin resistance + ARS_Mut-PETase_SP20His + ccA_MHETase_ SP20HA | Marlene Schlosser |
| BBa_K3002227 | Composite | L2 hygromycin resistance + ccA_Mut-PETase_SP20HA | Marlene Schlosser |
| BBa_K3002228 | Composite | L2 spectinomycin resistance + L1_connector_Pos2 + ccA_MHETase_SP20His | Marlene Schlosser |
| BBa_K3002229 | Composite | L2 hygromycin resistance + cCA_WT-PETase_SP20His | Marlene Schlosser |
| BBa_K3002230 | Composite | L2 spectinomycin resistance + cCA_WT-PETase_SP20His + cCA_MHETase_SP20HA | Marlene Schlosser |
| BBa_K3002231 | Composite | L2 hygromycin resistance + WT-PETase_His | Marlene Schlosser |
| BBa_K3002232 | Composite | L2 spectinomycin resistance + cCA_MutateM_SP20HA | Marlene Schlosser |
| BBa_K3002233 | Composite | L2 spectinomycin resistance + MutateM_HA | Marlene Schlosser |
| BBa_K3002234 | Composite | L2 hygromycin resistance 2 + cCA_Mut-PETase_SP20HA | Marlene Schlosser |
Level 1 Parts
Level 1 parts are functional units which can be used to form level 2 constructs. Those parts include at least a promoter, a coding sequence and a terminator. Optional are secretion signals, glycosylation tags and tags for protein detection which can only be found in level 1 parts with enzymes. As level 1 composite parts, we offer you two different antibiotic resistance cassettes and different combinations of enzymes with tags, secretion signals and glycosylation tags.
Level 2 Constructs
Overview of all level 2 MoClo constructs.
Level 2 constructs are a combination of level 1 constructs. A gene cluster can be created by ligating several genes in a row. This allows the transformation of a gene cluster into the chassis and avoids time consuming alternatives such as co-transformations. Depending on the vector of the level 1 part, the final position of the part in a level 2 ligation will already be determined. Each vector allows the creation of different sticky overhangs after digestion. A level 2 construct is composed of eight level 1 positions. If the final construct consists of less then eight level 1 parts, which is normally the case, a linker is needed to connect the sticky end of the level 1 part and the vector. Every position has its own unique overhang and therefore a linker exists for every possible number of level 1 constructs. The position of each level one construct is already determined by the choice of the vector which allows single tube ligations.
Best Composite Part
Our best composite part is the expression construct for spectinomycin resistance. It can be ligated to a level 2 construct with the desired level 1 constructs, thereby ensuring that the transformed cells can be selected. Transformants with the spectinomycin resistance cassette are plated on agar plates containing this antibiotic. Since the selection of transformants is a critical step in any further analysis of expressed constructs, we offer it to the iGEM community to simplify their recombinant research with Chlamydomonas reinhardtii.
