Parts

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Parts CISS iGEM 2025

Parts for E.Coli:

Type Part Link Name Functional Description
Composite (Device) BBa_25D3UKXW PET to TPA & EG using LCCICCG in E.coli Indicuble expression under the T7 promoter of the modified enzyme LCCICCG (leaf-branch compost cutinase) - https://parts.igem.org/Part:BBa_K3478888 GFP is also under the same promoter allowing for measurements of expression and optimization of conditions to maximize enzyme production. Biotechnology applications exist for LCCICCG to convert PET into TPA which can be further metabolised into higher value products (Sadler and Wallace, 2021).
Composite (Device) BBa_2522T7A8 TPA transporter in E.coli TPA transporter under constitutive promoter for E.coli. Contains the tpaK gene (https://parts.igem.org/Part:BBa_K4728011) originating from the species Rhodococcus jostii, tpaK encodes for the terephthalate transporter used during uptake. This transport protein allows for the transportation of terephthalic acid into the bacterial cells. This part can allow for combination with TPA metabolizing enzymes for upcycling of TPA into products such as Vanillin (Sadler and Wallace, 2021 DOI:https://doi.org/10.1039/D1GC00931A) or PCA.
Composite (Device) BBa_25CF2NOP TPA to PCA in E.coli This part allows for the uptake of Terephthalate (TPA) in E.coli and concurrently for the metabolism of TPA into PCA (protocatechuate). This is designed to be an essential step in the biotechnological process of upcycling PET plastic into higher value products. This composite part uses basic parts coding for genes tphA1, 2, 3, and DCDDH which produce enzymes involved in the conversion of TPA to PCA.
Basic BBa_25VDWFRT pcaGH - Protocatechuate 3,4-dioxygenase beta and alpha subunits Pseudomonas putida (ATCC 23975) protocatechuate 3,4-dioxygenase beta and alpha subunits (pcaH and pcaG) genes, complete cds.. Codes for the enzyme Protocatechuate 3,4-dioxygenase, part of the beta-ketoadipate pathway. Catalyzes the production of 3-carboxy-cis,cis-muconate from 3,4-dihydroxybenzoate.
Basic BBa_251AN6OX pcaIJ – beta-ketoadipate: succinyl-coA transferase Pseudomonas putida beta-ketoadipate: succinyl-coA transferase (pcaI, pcaJ) genes, complete cds.
Basic BBa_25JXTXSM pcaB - 3-carboxy-cis,cis-muconate cycloisomerase (pcaB) gene b-Carboxy-cis,cis-muconate lactonizing enzyme (cycloisomerase) from P. putida. Part of the beta-ketoadipate pathway; Facilitating the reaction: 5-oxo-4,5-dihydro-2-furylacetate from 3-carboxy-cis,cis-muconate.
Basic BBa_256JN97U PcaF - Beta-ketoadipyl-CoA thiolase PcaF gene from Pseudomonas putida KT2440. Catalyzes thiolytic cleavage of beta-ketoadipyl-CoA to succinyl-CoA and acetyl-CoA.
Basic BBa_25T1AMVB PCA (protocatechuate) Transporter 4-hydroxybenzoate transporter PcaK from Pseudomonas aeruginosa. Transports 4-hydroxybenzoate (4-HBA) and protocatechuate across the membrane. Driven by the proton motive force (https://www.uniprot.org/uniprotkb/Q9I6Q3/entry).
Composite (Device) BBa_25VAG5KT PCA transporter in E.coli TPA transporter under constitutive promoter for E.coli. Contains the pcaK gene (https://registry.igem.org/parts/bba-25t1amvb) originating from the species Pseudomonas aeruginosa, pcaK transports 4-hydroxybenzoate (4-HBA) and protocatechuate across the membrane. Driven by the proton motive force. (https://www.uniprot.org/uniprotkb/Q9I6Q3/entry)
Composite (Device) BBa_254GSRYG Protocatechuate (PCA) to Acetyl CoA in E.coli Conversion of PCA to Acetyl CoA via several enzymes from the beta-ketoadipate pathway. The system is inducible under the T7 promoter for string expression in E.coli.
Composite (Device) BBa_25GP9EZL PHB production and optimization with ZWF in E.coli For the inducible (T7) production of Poly-3-hydroxybutyrate, (PHB) in E.coli, a type of Polyhydroxyalkanoate (PHA) which is a biodegradable plastic which accumulates inside cells. PHB is produced via the biosynthesis of 3 enzymes encoded by the genes phaA, phaB and phaC. Optimized with the ZWF gene which codes for Glucose-6-phosphate 1-dehydrogenase, which Catalyzes the oxidation of glucose 6-phosphate to 6-phosphogluconolactone. The purpose of this gene is to increase the amount of NADPH available (Lim et al., 2002) and thus upregulate the PHB biosynthesis pathway.
Composite (Device) BBa_25HZF1BX PHB production and optimization with SerA in E.coli For the inducible (T7) production of Poly-3-hydroxybutyrate, (PHB) in E.coli, a type of Polyhydroxyalkanoate (PHA) which is a biodegradable plastic which accumulates inside cells. PHB is produced via the biosynthesis of 3 enzymes encoded by the genes phaA, phaB and phaC. To up-regulate the biosynthesis pathway of PHB production, this part also contains Serine Repeat Antigen (SerA) gene (https://parts.igem.org/Part:BBa_K2086001). This gene codes for the protein 3-phosphoglycerate dehydrogenase, also known as PHGDH. The purpose of this gene is to increase the amount of NADPH available (Zhang et al., 2014) and thus upregulate the PHB biosynthesis pathway.
Composite (Device) BBa_254YY4J3 Polyhydroxybutyrate (PHB) depolymerase under T7 promoter with GFP Under inducible expression via the T7 promoter, the extracellular poly(3-hydroxybutyrate) (PHB) depolymerase PhaZ - isolated from Penicillium funiculosum, can be recombinantly expressed in E. coli cells. The enzyme is a glycoprotein composed of a single polypeptide chain with a molecular mass of about 37,000 Da. GFP is fused to the enzyme to measure expression levels. Brucato CL, Wong SS. Extracellular poly(3-hydroxybutyrate) depolymerase from Penicillium funiculosum: general characteristics and active site studies. Arch Biochem Biophys. 1991 Nov 1;290(2):497-502. doi: 10.1016/0003-9861(91)90572-z. PMID: 1929416.

Parts for Pseudomonas Putida

Type Part Link Name Functional Description
Basic BBa_25N4U2VJ RBS for Pseudomonas putida To improve the P. putida toolkit, Elmore et al., 2017 created a library of RBS and promoters for this strain. RBS JER07 showed protein production levels varied by ~6–7-fold from the highest to lowest strength RBS when optimized with various promoter variants. https://pmc.ncbi.nlm.nih.gov/articles/PMC5699527/#s0045
Basic BBa_251KQA88 Poly(3-hydroxyalkanoate) polymerase subunit PhaC with RBS for P. putida When expressed in conjunction with the genes PhaA and PhaB (from C.necator), this confers the ability to synthesize up to 13% (w/w) poly(3-hydroxybutyrate) (PHB) depending on the carbon source; all 4 genes are necessary for PHB production (https://www.uniprot.org/uniprotkb/P73390/entry). This coding sequence is combined with an RBS sequence (https://registry.igem.org/parts/bba-25n4u2vj) for translation within Pseudomonas putida - this provides an alternative to expressing in E. coli as described by previous iGEM teams (https://parts.igem.org/Part:BBa_K934001)
Basic BBa_2562DM8W Acetyl-CoA acetyltransferase - PhaA with RBS for P. putida Acetyl-CoA acetyltransferase - PhaA from Cupriavidus necator. Catalyzes the condensation of two acetyl-coA units to form acetoacetyl-CoA. Involved in the biosynthesis of polyhydroxybutyrate (PHB), accumulated as an intracellular energy reserve material when cells grow under nutrient limitation. https://www.uniprot.org/uniprotkb/P14611/entry. This coding sequence is combined with an RBS sequence (https://registry.igem.org/parts/bba-25n4u2vj) for translation within P. putida - this provides an alternative to expressing in E. coli as described by previous iGEM teams (https://parts.igem.org/Part:BBa_K934001)
Basic BBa_25I7O4MN Acetoacetyl-CoA reductase - PhaB with RBS for P. putida Catalyzes the chiral reduction of acetoacetyl-CoA to (R)-3-hydroxybutyryl-CoA. Source from Cupriavidus necator. Is involved in the biosynthesis of polyhydroxybutyrate (PHB), which is accumulated as an intracellular energy reserve material when cells grow under conditions of nutrient limitation https://www.uniprot.org/uniprotkb/P14697/entry This coding sequence is combined with an RBS sequence (https://registry.igem.org/parts/bba-25n4u2vj) for translation within Pseudomonas putida - this provides an alternative to expressing in E. coli as described by previous iGEM teams (https://parts.igem.org/Part:BBa_K934001)
Composite (Device) BBa_250E79FK PET to TPA in P. putida Inducible PETase LCC-ICCG (Leaf-branch compost cutinase), a highly thermostable PETase enzyme capable of high levels of Polyethylene terephthalate (PET) degradation (https://parts.igem.org/Part:BBa_K3478888). Designed to be used in P. putida under the Xyls-Pm expression system.
Composite (Device) BBa_25T2Z1R8 PHB production up-regulation with SerA in P. putida This part is designed to be used in Pseudomonas putida for the production of Poly-3-hydroxybutyrate, (PHB), a type of Polyhydroxyalkanoate (PHA), a biodegradable plastic which accumulates inside cells. PHB is produced via the biosynthesis of 3 enzymes encoded by the genes phaA, phaB and phaC. To up-regulate the biosynthesis pathway of PHB production, this part also contains Serine Repeat Antigen (SerA) gene (https://parts.igem.org/Part:BBa_K2086001). This gene codes for the protein 3-phosphoglycerate dehydrogenase, also known as PHGDH. The purpose of this gene is to increase the amount of NADPH available (Zhang et al., 2014) and thus upregulate the PHB biosynthesis pathway. We incorporated the Xysl/Pm (https://parts.igem.org/Part:BBa_K1973014) expression system which is recommended for P. putida. (Gawin et al., 2017)
Composite (Device) BBa_258G509X PHB production up-regulation with ZWF in P. putida This part is designed to be used in Pseudomonas putida for the production of Poly-3-hydroxybutyrate, (PHB), a type of Polyhydroxyalkanoate (PHA), a biodegradable plastic which accumulates inside cells. PHB is produced via the biosynthesis of 3 enzymes encoded by the genes phaA, phaB and phaC. To up-regulate the biosynthesis pathway of PHB production, this part also contains the ZWF gene (https://parts.igem.org/Part:BBa_K1674004). This gene codes for the protein Glucose-6-Phosphate-Dehydrogenase. The purpose of this gene is to increase the amount of NADPH available (Lim et al., 2002) and thus upregulate the PHB biosynthesis pathway. We incorporated the Xysl/Pm (https://parts.igem.org/Part:BBa_K1973014) expression system which is recommended for P. putida. (Gawin et al., 2017)