Team:DTU-Denmark/Composite Part

Composite parts

We made composite parts

Type: Regulatory

Size: 683 bp

This is a strong promoter for Aspergillus niger that has high activity in the exponential phase. This is a synthetic exponential phase promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the glaA promoter from different Aspergillus spp., and the gene was chosen based on RNA-seq data from Aspergillus niger. The consensus promoter was expected to have really high expression. This version has “noise” added to the sequence to support a hypothesis that it was possible to turn down expression of the promoter.

The figure shows the 683 basepair long promoter PLEAPglaA_2

Assembly Compatibility:

The figure shows the compatibility of PLEAPglaA_2

Type: Regulatory

Size: 842 bp

This is a medium strength constitutive promoter for Aspergillus niger. This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the sonB promoter from different Aspergillus spp., and the gene was chosen based on RNA-seq data from Aspergillus niger. [1] This version is a consensus sequence of the sonB promoters in Aspergillus and is expected to have a medium strength expression.

The figure shows 842 basepair long promoter PLEAPsonB_1

Assembly Compatibility:

The figure shows the compatibility of promoter PLEAPsonB_1

Type: Regulatory

Size: 926 bp

This is a strong constitutive promoter for Aspergillus niger that has especially high activity in the lag phase. This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the gpdA promoter from different Aspergillus spp., and the gene was chosen based on RNA-seq data from Aspergillus niger. This version is a consensus sequence of the promoters in Aspergillus and is expected to have a high expression.

The figure shows the 926 basepair long promoter PLEAPgdpA_1

Assembly Compatibility:

The figure shows the compatibility of PLEAPgpdA_1

Type: Regulatory

Size: 926 bp

This is a medium strength constitutive promoter for Aspergillus niger. This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the gpdA promoter from different Aspergillus spp., and the gene was chosen based on RNA-seq data from Aspergillus niger. The consensus promoter was expected to have high expression. This version has “noise” added to the sequence to support a hypothesis that it was possible to turn down expression of the promoter.

The figure shows the 926 basepair long promoter PLEAPgdpA_2

Assembly Compatibility:

The figure shows the compatibility of PLEAPgpdA_2

Type: Regulatory

Size: 986 bp

This is a very strong constitutive promoter for Aspergillus niger that has especially high activity in the exponential growth phase. This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the mstA promoter from different Aspergillus spp., and the gene was chosen based on RNA-seq data from Aspergillus niger. This version is a consensus sequence of the mstA promoters in Aspergillus and is expected to have a high constitutive expression.

The figure shows the 986 basepair long promoter PLEAPmstA_1

Assembly Compatibility:

The figure shows the compatibility of PLEAPmstA_1

Type: Regulatory

Size: 933 bp

This is a weak constitutive promoter for Aspergillus niger. This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on an unknown promoter from different Aspergillus spp., and the gene was chosen based on RNA-seq data from Aspergillus niger. This version is a consensus sequence of the An07g08210 promoters in Aspergillus and is expected to have a weak constitutive expression.

The figure shows the 933 basepair long promoter PLEAPunk_1

Assembly Compatibility:

The figure shows the compatibility of PLEAPunk_1

Type: Regulatory

Size: 973 bp

This is a strong promoter for Aspergillus niger that has high activity in the late exponential phase and stationary phase. This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the gfaA promoter from different Aspergillus spp., and the gene was chosen based on RNA-seq data from Aspergillus niger. This version is a consensus sequence of the promoters in Aspergillus and is expected to have a high expression.

The figure shows the 973 basepair long promoter PLEAPgfaA_1

Assembly Compatibility:

The figure shows the compatibility of PLEAPgfaA_1

Type: Regulatory

Size: 1014 bp

This is a strong promoter for Aspergillus niger that has high activity in the stationary growth phase. This is a synthetic stationary phase promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the hfbD promoter from different Aspergillus spp., and the gene was chosen based on RNA-seq data from Aspergillus niger. This is a consensus promoter and are expected to have high expression in the stationary phase.

The figure shows the 1014 basepair long promoter PLEAPhfbD_1

Assembly Compatibility:

The figure shows the compatibility of PLEAPhfbD_1

Type: Measurement

Size: 1505 bp

This is a device for testing promoters by expressing mCherry.

The figure shows the components of the fungal MoClo promoter test device

Assembly Compatibility:

The figure shows the compatibility of the fungal MoClo promoter test device

Type: Regulatory

Size: 683 bp

This is a promoter for Aspergillus niger of unknown strength that is expected to have high expression in the exponential growth phase. This is a synthetic exponential phase promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the glaA promoter from different Aspergillus spp., and the gene was chosen based on RNA-seq data from Aspergillus niger. The consensus promoter was expected to have really high expression. This version has “noise” added to the sequence to support a hypothesis that it was possible to turn down expression of the promoter.

The figure shows the 683 basepair long promoter PLEAPglaA_1

Assembly Compatibility:

The figure shows the compatibility of PLEAPglaA_1

Type: Regulatory

Size: 842 bp

This is a promoter for Aspergillus niger of unknown strength that is expected to have constitutive expression. This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the sonB promoter from different Aspergillus spp., and the gene was chosen based on RNA-seq data from Aspergillus niger. This version is a consensus sequence of the sonB promoters in Aspergillus and is expected to have a medium strength expression

The figure shows the 842 basepair long promoter PLEAPsonB_2

Assembly Compatibility:

The figure shows the compatibility of PLEAPsonB_2

Type: Regulatory

Size: 986 bp

This is a very strong constitutive promoter for Aspergillus niger that has especially high activity in the exponential growth phase. This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the gfaA promoter from different Aspergillus spp., and the gene was chosen based on RNA-seq data from Aspergillus niger. This version has introduced stokastic noise compared to the consensus sequence of the promoters in Aspergillus and is therefore expected to have a lower expression.

The figure shows the 986 basepair long promoter PLEAPmstA_2

Assembly Compatibility:

The figure shows the compatibility of PLEAPmstA_2

Type: Regulatory

Size: 724 bp

This is a promoter for Aspergillus niger of unknown strength that should be active in the stationary phase. This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the sonB promoter from different Aspergillus spp., and the gene was chosen based on RNA-seq data from Aspergillus niger. [1] This version is a consensus sequence of an unknown gene's promoters in Aspergillus.

The figure shows the 724 basepair long promoter PLEAPunk_4

Assembly Compatibility:

The figure shows the compatibility of PLEAPunk_4

Type: Regulatory

Size: 973 bp

This is a promoter on unknown for Aspergillus niger that is expected to have high activity in the late exponential phase and stationary phase. This is a synthetic constitutive promoter, created as part of the LEAP (Library of Engineered Aspergillus Promoters) project. It is based on the gfaA promoter from different Aspergillus spp., and the gene was chosen based on RNA-seq data from Aspergillus niger. This version has introduced stokastic noise compared to the consensus sequence of the promoters in Aspergillus and is therefore expected to have a lower expression.

The figure shows the 973 basepair long promoter PLEAPgfaA_2

Assembly Compatibility:

The figure shows the compatibility of PLEAPgfaA_2

Type: Regulatory

Size: 238 bp

This part is the bacterial promoter BBa_R0010 with MoClo level 0 compatible overhangs.

The figure shows the 238 basepair long MoClo promoter stand-in

Assembly Compatibility:

The figure shows the compatibility of the MoClo promoter stand-in

Type: Terminator

Size: 485 bp

Transcriptional terminator from Aspergillus nidulans. Domesticated to be in compliance with RFC[1000]

The figure shows the 485 basepair long Ttef (Domesticated from <i>Aspergillus nidulans</i>)

Assembly Compatibility:

The figure shows the compatibility of Ttef

Type: Tag

Size: 63 bp

Protein secretion tag from the GlaA enzyme from Aspergillus niger.

The figure shows the 63 basepair long Export signal from <i>A. niger</i> GlaA protein

Assembly Compatibility:

Compatibility of export signal from <i>A. niger</i> GlaA protein

Type: Coding

Size: 720 bp

Coming soon

The figure shows the 720 basepair long MoxGFP

Assembly Compatibility:

Compatibility of MoxGFP

Type: Measurement

Size: 10725 bp

This plasmid is designed as a shuttle vector to test different fungal promoters

The figure shows the 10725 basepair long promoter test plasmid pAMA1-zing_Pyr

Assembly Compatibility:

Compatibility of promoter test plasmid pAMA1-zing_Pyr

Type: Composite

Size: 1324 bp

This PyrG transcriptional unit can be used as a marker gene to select for succesful transformants of Pyr- mutants.

The figure shows the 1324 basepair long PyrG selection marker from <i>A. fumigatus</i>

Assembly Compatibility:

Compatibility of PyrG selection marker from <i>A. fumigatus</i>

Type: Plasmid Backbone

Size: 7896 bp

This is a plasmid containing an E.coli ORI, an ampicillin selection marker, and the AMA1 sequence. Inserts can be integrated into the plasmid by using a PacI/Nt.BbvCI USER assembly casette.

The figure shows the 7896 basepair long pAMA1_Amp_PacI-Nt.BbvCI plasmid backbone

Assembly Compatibility:

Compatibility of pAMA1_Amp_PacI-Nt.BbvCI plasmid backbone

Type: Coding

Size: 5721 bp

This sequence is used for creating autonomously replicating plasmids to use for Aspergillus spp.

The figure shows the 5721 basepair long AMA1 sequence for autonomous replication in <i>Aspergillus</i>

Assembly Compatibility:

Compatibility of AMA1 sequence for autonomous replication in <i>Aspergillus</i>

Type: Coding

Size: 906 bp

Coming soon

The figure shows the 906 basepair long PyrG CDS from <i>Aspergillus fumigatus</i>

Assembly Compatibility:

Compatibility of PyrG CDS from <i>Aspergillus fumigatus</i>

Type: Regulatory

Size: 418 bp

Promoter from the PyrG selection marker gene from Aspergillus fumigatus

The figure shows the 418 basepair long PyrG promoter from <i>Aspergillus fumigatus</i>

Assembly Compatibility:

Compatibility of PyrG promoter from <i>Aspergillus fumigatus</i>

Type: Composite

Size: 5546 bp

This part is designed to be inserted into pSB1C3 by Gibson cloning to make an integration plasmid for Aspergillus niger. The part is meant to be linearised by PCR prior to protoplast-mediated transformation, where the IS2 sites will lead to genomic integration of the insert (in this case just PyrG part BBa_K3046022) into the albA conidial pigment gene. Correct gene integration will, therefore, yield white/yellow conidia instead of the black conidia seen for the wildtype A. niger.

The figure shows the 5546 basepair long pGIA2P1 insert

Assembly Compatibility:

Compatibility of pGIA2P1 insert

Type: Measurement

Size: 7139 bp

This part is designed to be inserted into pSB1C3 by Gibson cloning to make an integration plasmid for Aspergillus niger. The part is meant to be linearised by PCR prior to protoplast-mediated transformation, where the IS2 sites will lead to genomic integration of the insert (in this case just PyrG part BBa_K3046022) into the albA conidial pigment gene. Correct gene integration will therefore yield white/yellow conidia instead of the black conidia seen for the wildtype A. niger.

The figure shows the 7139 basepair long pPEA2P1 insert

Assembly Compatibility:

Compatibility of pPEA2P1 insert

Type: DNA

Size: 2096 bp

This part is a long sequence of DNA that are homologous to a part of the albA conidial pigment gene. This homology can be used for homologous recombination of genomic constructs flanked by this IS2 sequence (along with the downstream sequence BBa_K3046033).

The figure shows the 2096 basepair long IS2 upstream sequence for <i>A. niger</i> genome integration

Assembly Compatibility:

Compatibility of IS2 upstream sequence for <i>A. niger</i> genome integration

Type: DNA

Size: 2112 bp

This part is a long sequence of DNA that are homologous to a part of the albA conidial pigment gene. This homology can be used for homologous recombination of genomic constructs flanked by this IS2 sequence (along with the upstream sequence BBa_K3046032).

The figure shows the 2112 basepair IS2 downstream sequence for <i>A. niger</i> genome integration

Assembly Compatibility:

Compatibility of IS2 downstream sequence for <i>A. niger</i> genome integration
The logos of our three biggest supporters, DTU Blue Dot, Novo Nordisk fonden and Otto Mønsted fonden The logos of all of our sponsors, DTU, BioNordica, Eurofins Genomics, Qiagen, NEB New England biolabs, IDT Integrated DNA technologies and Twist bioscience