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Vector Backbone Description: Backbone Marker:self-made; Backbone Size:2247; Vector Backbone:pUC19-derived; Vector Types:Unspecified; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:21364738
Comments: level 0 cloning vector
Proper citation: RRID:Addgene_47996 Copy
Vector Backbone Description: Backbone Marker:self-made; Backbone Size:2247; Vector Backbone:pUC19-derived; Vector Types:Unspecified; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:21364738
Comments: level 0 cloning vector
Proper citation: RRID:Addgene_47994 Copy
Vector Backbone Description: Backbone Marker:self-made; Backbone Size:2247; Vector Backbone:pUC19-derived; Vector Types:Unspecified; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:21364738
Comments: level 0 cloning vector
Proper citation: RRID:Addgene_47999 Copy
Vector Backbone Description: Backbone Marker:self-made; Backbone Size:2247; Vector Backbone:pUC19-derived; Vector Types:Unspecified; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:21364738
Comments: level 0 cloning vector
Proper citation: RRID:Addgene_47997 Copy
Vector Backbone Description: Backbone Marker:self-made; Backbone Size:2247; Vector Backbone:pUC19-derived; Vector Types:Unspecified; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:21364738
Comments: level 0 cloning vector
Proper citation: RRID:Addgene_47998 Copy
Vector Backbone Description: Backbone Marker:self-made; Backbone Size:4604; Vector Backbone:derived from pBIN19 and pUC19; Vector Types:Unspecified; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:21364738
Comments: level M cloning vector, replicates in E.coli and Agrobacterium, binary vector for Agrobacterium T-DNA delivery to plant cells
Proper citation: RRID:Addgene_48042 Copy
Vector Backbone Description: Backbone Marker:self-made; Backbone Size:4604; Vector Backbone:derived from pBIN19 and pUC19; Vector Types:Unspecified; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:21364738
Comments: level M cloning vector, replicates in E.coli and Agrobacterium, binary vector for Agrobacterium T-DNA delivery to plant cells
Proper citation: RRID:Addgene_48043 Copy
Vector Backbone Description: Backbone Marker:self-made; Backbone Size:2247; Vector Backbone:pUC19-derived; Vector Types:Unspecified; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:21364738
Comments: level 0 cloning vector
Proper citation: RRID:Addgene_47991 Copy
Vector Backbone Description: Backbone Marker:self-made; Backbone Size:2247; Vector Backbone:pUC19-derived; Vector Types:Unspecified; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:21364738
Comments: level 0 cloning vector
Proper citation: RRID:Addgene_47992 Copy
Vector Backbone Description: Backbone Marker:self-made; Backbone Size:2247; Vector Backbone:pUC19-derived; Vector Types:Unspecified; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:21364738
Comments: level 0 cloning vector
Proper citation: RRID:Addgene_47985 Copy
Vector Backbone Description: Backbone Marker:self-made; Backbone Size:4604; Vector Backbone:derived from pBIN19 and pUC19; Vector Types:Unspecified; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:21364738
Comments: level M cloning vector, replicates in E.coli and Agrobacterium, binary vector for Agrobacterium T-DNA delivery to plant cells
Proper citation: RRID:Addgene_48038 Copy
Vector Backbone Description: Backbone Marker:self-made; Backbone Size:2247; Vector Backbone:pUC19-derived; Vector Types:Unspecified; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:21364738
Comments: level 0 cloning vector
Proper citation: RRID:Addgene_47986 Copy
Vector Backbone Description: Backbone Marker:self-made; Backbone Size:3293; Vector Backbone:pUC19-derived; Vector Types:Unspecified; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:21364738
Comments: level 2 end linker
Proper citation: RRID:Addgene_48022 Copy
Vector Backbone Description: Backbone Marker:self-made; Backbone Size:3293; Vector Backbone:pUC19-derived; Vector Types:Unspecified; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:21364738
Comments: level 2 end linker
Proper citation: RRID:Addgene_48016 Copy
Vector Backbone Description: Backbone Marker:self-made; Backbone Size:3293; Vector Backbone:pUC19-derived; Vector Types:Unspecified; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:21364738
Comments: level 2 end linker
Proper citation: RRID:Addgene_48019 Copy
Species: M. jannaschii
Genetic Insert: polyspecific MJ tyrosyl synthetase
Vector Backbone Description: Backbone Marker:Schultz lab; Vector Backbone:pULTRA; Vector Types:Bacterial Expression; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:17061854
Comments: This plasmid was also used in Chatterjee et al Biochemistry. 2013 Feb 27. PMID: 23379331
Proper citation: RRID:Addgene_48215 Copy
Species: Homo sapiens
Genetic Insert: dCas9(D10A;H840A)
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:2799; Vector Backbone:pCR8/GW/TOPO; Vector Types:CRISPR, Gateway Cloning; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:23979020
Comments: For more information including protocols and
updates, please go to http://www.crispr-on.org
Proper citation: RRID:Addgene_48218 Copy
Species: Homo sapiens
Genetic Insert: dCas9(D10A;H840A) fusion with VP160 activation domain
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:2799; Vector Backbone:pCR8/GW/TOPO; Vector Types:CRISPR, Gateway Cloning; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:23979020
Comments: For more information including protocols and
updates, please go to http://www.crispr-on.org
Proper citation: RRID:Addgene_48221 Copy
Vector Backbone Description: Backbone Size:4785; Vector Backbone:pET; Vector Types:Bacterial Expression; Bacterial Resistance:Spectinomycin
Comments: This plasmid is an empty vector to be used with a LIC cloning protocol.
It has a TEV cleavable His6-MBP fusion tag on the N-terminus and a Spec resistance.
To clone into this vector, add LIC fusion tags to the 5' end of your PCR primers.
Forward - 5'TACTTCCAATCCAATGCA3'
Reverse - 5'TTATCCACTTCCAATGTTATTA3'
Linearize the plasmid with SspI and gel purify.
When digesting the DNA with T4 polymerase for LIC, use dCTP for insert and dGTP for vector. The 13-series vectors were designed to enable rapid cloning for a co-expression system. Vectors are based on Novagen's Duet system. Each gene is expressed on its own vector, which has been optimized so that each gene expresses at approximately equal levels. The 13-series vectors are all compatible with 2-series transfer vectors, so if cotransformation fails, there is a readily available backup in polycistronic expression.
13S CDF origin SpecR13K ColA origin KanR2-series transfer ColE1 origin AmpR13S vectors must be cotransformed with a 2-series vector for optimal expression (that is, the presence of an empty 2A-T vector enhances expression of a gene in 13S). For triple expressions, the proteins all seem to express at approximately equal levels. Genes in the CDF vector consistently express at very slightly lower levels, so if you're trying to pull down stoichiometric complexes, it's probably best to put His6-fusion protein in this vector. More information on this vector can be found through http://qb3.berkeley.edu/qb3/macrolab/
Proper citation: RRID:Addgene_48325 Copy
Vector Backbone Description: Backbone Size:3666; Vector Backbone:pET; Vector Types:Bacterial Expression; Bacterial Resistance:Spectinomycin
Comments: This plasmid is an empty vector to be used with a LIC cloning protocol.
It has a TEV cleavable His6-StrepII fusion tag on the N-terminus and a Spec resistance.
To clone into this vector, add LIC fusion tags to the 5' end of your PCR primers.
Forward - 5'TACTTCCAATCCAATGCA3'
Reverse - 5'TTATCCACTTCCAATGTTATTA3'
Linearize the plasmid with SspI and gel purify.
When digesting the DNA with T4 polymerase for LIC, use dCTP for insert and dGTP for vector. The 13-series vectors were designed to enable rapid cloning for a co-expression system. Vectors are based on Novagen's Duet system. Each gene is expressed on its own vector, which has been optimized so that each gene expresses at approximately equal levels. The 13-series vectors are all compatible with 2-series transfer vectors, so if cotransformation fails, there is a readily available backup in polycistronic expression.
13S CDF origin SpecR13K ColA origin KanR2-series transfer ColE1 origin AmpR13S vectors must be cotransformed with a 2-series vector for optimal expression (that is, the presence of an empty 2A-T vector enhances expression of a gene in 13S). For triple expressions, the proteins all seem to express at approximately equal levels. Genes in the CDF vector consistently express at very slightly lower levels, so if you're trying to pull down stoichiometric complexes, it's probably best to put His6-fusion protein in this vector. More information on this vector can be found through http://qb3.berkeley.edu/qb3/macrolab/
Proper citation: RRID:Addgene_48326 Copy
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