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moclo tool kit  (Addgene inc)


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    Addgene inc moclo tool kit
    Moclo Tool Kit, supplied by Addgene inc, used in various techniques. Bioz Stars score: 95/100, based on 84 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/moclo+tool+kit/MoClo+Toolkit+(Kit+%231000000044)/pmc12868398-157-10-15
    Average 95 stars, based on 84 article reviews
    moclo tool kit - by Bioz Stars, 2026-09
    95/100 stars

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    Related Articles

    Cloning:

    Article Title: Tm-2 2 resistance targets a conserved cysteine essential for tobacco mosaic virus (TMV) movement
    Article Snippet: .. Plasmids p35S:Tm-2 2 , p35S:MP TMV , p35S:MP ToBRFV , p35S:MP TMV -YFP + ERmCherry and p35S:MP ToBRFV -YFP + ER-mCherry were assembled by Golden Gate cloning using the MoClo tool kit for plants (Addgene) ( ) as described ( ). ..

    Article Title: Activation of Tm‐2 2 resistance is mediated by a conserved cysteine essential for tobacco mosaic virus movement
    Article Snippet: .. Plasmids p35S :Tm‐2 2 , p35S: MP TM V , p35S: MP ToBRFV , p35S: MP TM V ‐YFP + ER‐mCherry and p35S:MP ToBRFV ‐YFP + ER‐mCherry were assembled by Golden Gate cloning using the MoClo tool kit for plants (Addgene) (Weber et al., ) as described by Hak and Spiegelman ( ). ..

    Article Title: Arogenate dehydrogenase polynucleotides, polypeptides and methods of using the same
    Article Snippet: Transcripts were amplified by PCR using gene specific primers (Table S1) and Phusion High-Fidelity DNA polymerase (Thermo Fisher Scientific, Waltham, MA, USA). .. Vectors for transient transformation were constructed with Golden Gate cloning using the MoClo Tool Kit (Weber et al., 2011; Addgene, Cambridge, MA, USA), with the BpiI and BsaI restriction sites eliminated after cloning. .. The turboGFP sequence used in this assay was a variant codon-optimized for plants contained in the MoClo Plant Parts Kit (Engler et al., 2014; Addgene, Cambridge, MA, USA).

    Article Title: Activation of Tm-2 2 resistance is mediated by a conserved cysteine essential for tobacco mosaic virus movement.
    Article Snippet: .. Plasmids p35S:Tm- 22, p35S:MPTMV, p35S:MPToBRFV, p35S:MPTMVYFP + ER- mCherry and p35S:MPToBRFV- YFP + ER- mCherry were assembled by Golden Gate cloning using the MoClo tool kit for plants (Addgene) (Weber et al., 2011) as described by Hak and Spiegelman (2021). ..

    Article Title: Genome editing in almond: A CRISPR-based approach through hairy root transformation
    Article Snippet: .. Plasmid construction was performed by modular cloning using the MoClo Tool Kit and the MoClo Plant Parts Kit (Addgene, – ). ..

    Transformation Assay:

    Article Title: Pyrroline-5-carboxylate metabolism protein complex detected in Arabidopsis thaliana leaf mitochondria.
    Article Snippet: .. Transformation of Arabidopsis with transgenic GFP-fusion constructs The cDNA sequence of Arabidopsis P5CDH was modified by sitedirected mutagenesis to remove the BsaI restriction site, and cloned with the addition of an enterokinase recognition and cleavage site into the pAGM1287 entry plasmid (MoClo Tool Kit, Addgene) (Weber et al., 2011) using the primers described in Supplementary Table S1. ..

    Article Title: Arogenate dehydrogenase polynucleotides, polypeptides and methods of using the same
    Article Snippet: Transcripts were amplified by PCR using gene specific primers (Table S1) and Phusion High-Fidelity DNA polymerase (Thermo Fisher Scientific, Waltham, MA, USA). .. Vectors for transient transformation were constructed with Golden Gate cloning using the MoClo Tool Kit (Weber et al., 2011; Addgene, Cambridge, MA, USA), with the BpiI and BsaI restriction sites eliminated after cloning. .. The turboGFP sequence used in this assay was a variant codon-optimized for plants contained in the MoClo Plant Parts Kit (Engler et al., 2014; Addgene, Cambridge, MA, USA).

    Transgenic Assay:

    Article Title: Pyrroline-5-carboxylate metabolism protein complex detected in Arabidopsis thaliana leaf mitochondria.
    Article Snippet: .. Transformation of Arabidopsis with transgenic GFP-fusion constructs The cDNA sequence of Arabidopsis P5CDH was modified by sitedirected mutagenesis to remove the BsaI restriction site, and cloned with the addition of an enterokinase recognition and cleavage site into the pAGM1287 entry plasmid (MoClo Tool Kit, Addgene) (Weber et al., 2011) using the primers described in Supplementary Table S1. ..

    Construct:

    Article Title: Pyrroline-5-carboxylate metabolism protein complex detected in Arabidopsis thaliana leaf mitochondria.
    Article Snippet: .. Transformation of Arabidopsis with transgenic GFP-fusion constructs The cDNA sequence of Arabidopsis P5CDH was modified by sitedirected mutagenesis to remove the BsaI restriction site, and cloned with the addition of an enterokinase recognition and cleavage site into the pAGM1287 entry plasmid (MoClo Tool Kit, Addgene) (Weber et al., 2011) using the primers described in Supplementary Table S1. ..

    Article Title: Arogenate dehydrogenase polynucleotides, polypeptides and methods of using the same
    Article Snippet: Transcripts were amplified by PCR using gene specific primers (Table S1) and Phusion High-Fidelity DNA polymerase (Thermo Fisher Scientific, Waltham, MA, USA). .. Vectors for transient transformation were constructed with Golden Gate cloning using the MoClo Tool Kit (Weber et al., 2011; Addgene, Cambridge, MA, USA), with the BpiI and BsaI restriction sites eliminated after cloning. .. The turboGFP sequence used in this assay was a variant codon-optimized for plants contained in the MoClo Plant Parts Kit (Engler et al., 2014; Addgene, Cambridge, MA, USA).

    Sequencing:

    Article Title: Pyrroline-5-carboxylate metabolism protein complex detected in Arabidopsis thaliana leaf mitochondria.
    Article Snippet: .. Transformation of Arabidopsis with transgenic GFP-fusion constructs The cDNA sequence of Arabidopsis P5CDH was modified by sitedirected mutagenesis to remove the BsaI restriction site, and cloned with the addition of an enterokinase recognition and cleavage site into the pAGM1287 entry plasmid (MoClo Tool Kit, Addgene) (Weber et al., 2011) using the primers described in Supplementary Table S1. ..

    Modification:

    Article Title: Pyrroline-5-carboxylate metabolism protein complex detected in Arabidopsis thaliana leaf mitochondria.
    Article Snippet: .. Transformation of Arabidopsis with transgenic GFP-fusion constructs The cDNA sequence of Arabidopsis P5CDH was modified by sitedirected mutagenesis to remove the BsaI restriction site, and cloned with the addition of an enterokinase recognition and cleavage site into the pAGM1287 entry plasmid (MoClo Tool Kit, Addgene) (Weber et al., 2011) using the primers described in Supplementary Table S1. ..

    Mutagenesis:

    Article Title: Pyrroline-5-carboxylate metabolism protein complex detected in Arabidopsis thaliana leaf mitochondria.
    Article Snippet: .. Transformation of Arabidopsis with transgenic GFP-fusion constructs The cDNA sequence of Arabidopsis P5CDH was modified by sitedirected mutagenesis to remove the BsaI restriction site, and cloned with the addition of an enterokinase recognition and cleavage site into the pAGM1287 entry plasmid (MoClo Tool Kit, Addgene) (Weber et al., 2011) using the primers described in Supplementary Table S1. ..

    Clone Assay:

    Article Title: Pyrroline-5-carboxylate metabolism protein complex detected in Arabidopsis thaliana leaf mitochondria.
    Article Snippet: .. Transformation of Arabidopsis with transgenic GFP-fusion constructs The cDNA sequence of Arabidopsis P5CDH was modified by sitedirected mutagenesis to remove the BsaI restriction site, and cloned with the addition of an enterokinase recognition and cleavage site into the pAGM1287 entry plasmid (MoClo Tool Kit, Addgene) (Weber et al., 2011) using the primers described in Supplementary Table S1. ..

    Article Title: A modular synthetic biology toolkit unlocks metabolic engineering of the industrially relevant alga Nannochloropsis
    Article Snippet: For plasmid construction and assembly, a molar ratio of 1:3 (insert-to-vector) was routinely applied. .. The parts were cloned into the level 0 vectors of MoClo Tool Kit (Kit #1000000044; Addgene). ..

    Article Title: A modular synthetic biology toolkit unlocks metabolic engineering of the industrially relevant alga Nannochloropsis.
    Article Snippet: For plasmid construction and assembly, a molar ratio of 1:3 (insert-to-vector) was routinely applied. .. The parts were cloned into the level 0 vectors of MoClo Tool Kit (Kit #1000000044; Addgene). ..

    Plasmid Preparation:

    Article Title: Pyrroline-5-carboxylate metabolism protein complex detected in Arabidopsis thaliana leaf mitochondria.
    Article Snippet: .. Transformation of Arabidopsis with transgenic GFP-fusion constructs The cDNA sequence of Arabidopsis P5CDH was modified by sitedirected mutagenesis to remove the BsaI restriction site, and cloned with the addition of an enterokinase recognition and cleavage site into the pAGM1287 entry plasmid (MoClo Tool Kit, Addgene) (Weber et al., 2011) using the primers described in Supplementary Table S1. ..

    Article Title: Genome editing in almond: A CRISPR-based approach through hairy root transformation
    Article Snippet: .. Plasmid construction was performed by modular cloning using the MoClo Tool Kit and the MoClo Plant Parts Kit (Addgene, – ). ..



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    Image Search Results


    A) Plasmids support cloning, library propagation, and gene expression, but unintended interactions with host transcriptional and replication machinery can drive recombination, plasmid instability, low plasmid DNA (pDNA) yields, and reduced expression. B) Overview of the pooled plasmid fitness assay. A diverse library of mammalian expression plasmid parts is assembled using Golden Gate cloning and transformed into E. coli . The pooled culture is grown over time, and changes in plasmid abundance are measured via long read nanopore sequencing and alignment. Log fold change (LFC) in relative plasmid abundance is used as a fitness metric and ranked across the library. Machine learning models predict sequence-dependent fitness effects. C) A set of Mammalian Toolkit (MTK) plasmids was transformed into E. coli DH5α, DH10B, and Stbl3 and the pooled assay performed in triplicate. Violin plots show plasmid fitness with three distinct categories of poor plasmid propagation including reduced fitness (light grey), low fitness (grey) and severe plasmid loss. This was compared to plasmid size (left) and GC content (right).

    Journal: bioRxiv

    Article Title: Quantitative profiling of millions of nucleotides reveals sequence-encoded interactions that govern plasmid propagation

    doi: 10.64898/2025.12.15.694402

    Figure Lengend Snippet: A) Plasmids support cloning, library propagation, and gene expression, but unintended interactions with host transcriptional and replication machinery can drive recombination, plasmid instability, low plasmid DNA (pDNA) yields, and reduced expression. B) Overview of the pooled plasmid fitness assay. A diverse library of mammalian expression plasmid parts is assembled using Golden Gate cloning and transformed into E. coli . The pooled culture is grown over time, and changes in plasmid abundance are measured via long read nanopore sequencing and alignment. Log fold change (LFC) in relative plasmid abundance is used as a fitness metric and ranked across the library. Machine learning models predict sequence-dependent fitness effects. C) A set of Mammalian Toolkit (MTK) plasmids was transformed into E. coli DH5α, DH10B, and Stbl3 and the pooled assay performed in triplicate. Violin plots show plasmid fitness with three distinct categories of poor plasmid propagation including reduced fitness (light grey), low fitness (grey) and severe plasmid loss. This was compared to plasmid size (left) and GC content (right).

    Article Snippet: Single-part plasmid libraries were assembled using the Mammalian Tool Kit (MTK) modular cloning framework, sourced from Addgene. ( ) Individual regulatory elements (promoters, UTRs, coding sequences, localization tags, and polyadenylation sequences) were cloned into a common bacterial backbone containing a chloramphenicol resistance marker and a pMB1-derived origin of replication.

    Techniques: Cloning, Gene Expression, Plasmid Preparation, Expressing, Transformation Assay, Nanopore Sequencing, Sequencing

    A) Four-part expression cassette assembly from the MTK library, comprising a promoter, coding sequence (CDS), localisation tag, and terminator, combined by one-pot Golden Gate cloning and propagated in E. coli DH5α. Pooled fitness was measured by quantifying plasmid abundance over a 16-hour batch culture. B) Plasmid log fold change (LFC) plotted against plasmid size (top) and GC content (bottom) for 1,847 assembled constructs. Each point represents one plasmid. C) Overview of the random forest regression workflow used to predict plasmid fitness from one-hot-encoded part identities. D) Predicted versus measured plasmid LFC for the random forest model. Each point represents one plasmid. E) Comparison of mean part fitness (LFC) with contextual effect estimated by mean SHAP value for each part. Points are coloured by cassette position (promoter, CDS, tag, terminator). F) SHAP value distributions showing part contributions to predicted plasmid fitness across all assemblies. Each ridge represents one part. G) Pairwise SHAP interaction values between selected parts. Points represent individual assemblies; distributions summarise interaction variability across the library.

    Journal: bioRxiv

    Article Title: Quantitative profiling of millions of nucleotides reveals sequence-encoded interactions that govern plasmid propagation

    doi: 10.64898/2025.12.15.694402

    Figure Lengend Snippet: A) Four-part expression cassette assembly from the MTK library, comprising a promoter, coding sequence (CDS), localisation tag, and terminator, combined by one-pot Golden Gate cloning and propagated in E. coli DH5α. Pooled fitness was measured by quantifying plasmid abundance over a 16-hour batch culture. B) Plasmid log fold change (LFC) plotted against plasmid size (top) and GC content (bottom) for 1,847 assembled constructs. Each point represents one plasmid. C) Overview of the random forest regression workflow used to predict plasmid fitness from one-hot-encoded part identities. D) Predicted versus measured plasmid LFC for the random forest model. Each point represents one plasmid. E) Comparison of mean part fitness (LFC) with contextual effect estimated by mean SHAP value for each part. Points are coloured by cassette position (promoter, CDS, tag, terminator). F) SHAP value distributions showing part contributions to predicted plasmid fitness across all assemblies. Each ridge represents one part. G) Pairwise SHAP interaction values between selected parts. Points represent individual assemblies; distributions summarise interaction variability across the library.

    Article Snippet: Single-part plasmid libraries were assembled using the Mammalian Tool Kit (MTK) modular cloning framework, sourced from Addgene. ( ) Individual regulatory elements (promoters, UTRs, coding sequences, localization tags, and polyadenylation sequences) were cloned into a common bacterial backbone containing a chloramphenicol resistance marker and a pMB1-derived origin of replication.

    Techniques: Expressing, Sequencing, Cloning, Plasmid Preparation, Construct, Comparison