Team:TU Kaiserslautern/Composite Part

Composite Parts

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.

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