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Species: Homo sapiens
Genetic Insert: cyclin B1
Vector Backbone Description: Backbone Marker:unknown; Vector Backbone:pVenus-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:25217616
Proper citation: RRID:Addgene_61851 Copy
Species: Homo sapiens
Genetic Insert: Cyclin B1
Vector Backbone Description: Backbone Marker:unknown; Vector Backbone:pCerulean N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Proper citation: RRID:Addgene_61852 Copy
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21925267
Comments: Genotype = cyo::kan++ ilvE (Kan cassette in the chromosomal DNA)
Precursor strain = CLY (Genotype cyo::kan)
Selective amino acid labeling (and/or requirement) = Ile, Leu*
CLY strain was derived from the C43(DE3)strain by transferring the cyo deletion (with kanamycin resistance cassette kanR) from an E. coli B strain to C43(DE3) strain by phage P1 [J. Biol. Chem. 282, 8777-8785 (2007)]. Thus, the kanR casette CANNOT be removed because there are NO FRT sites flanking kanR.
In the presence of 0.4-1 mM Tyr, tyrB is repressed and Leu is required for growth in minimal medium. This strategy can only be applicable for a short-term cultivation but not suitable for a long-term cultivation for heterologous expression of foreign genes.
Each target gene was deleted from the chromosome of C43 (DE3) E. coli strain using λ-Red recombination system.
Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
Proper citation: RRID:Addgene_61906 Copy
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21925267
Comments: Genotype = cyo::kan hisG (Kan cassette in the chromosomal DNA)
Precursor strain = CLY (Genotype cyo::kan)
Selective amino acid labeling (and/or requirement) = His
CLY strain was derived from the C43(DE3)strain by transferring the cyo deletion (with kanamycin resistance cassette kanR) from an E. coli B strain to C43(DE3) strain by phage P1 [J. Biol. Chem. 282, 8777-8785 (2007)]. Thus, the kanR casette CANNOT be removed because there are NO FRT sites flanking kanR.
Each target gene was deleted from the chromosome of C43 (DE3) E. coli strain using λ-Red recombination system.
Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
Proper citation: RRID:Addgene_61907 Copy
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21925267
Comments: Genotype = cyo::kan++ ilvE avtA (Kan cassette in the chromosomal DNA)
Precursor strain = ML2 (Genotype cyo::kan)
Selective amino acid labeling (and/or requirement) = Ile, Leu*, Val
CLY strain was derived from the C43(DE3)strain by transferring the cyo deletion (with kanamycin resistance cassette kanR) from an E. coli B strain to C43(DE3) strain by phage P1 [J. Biol. Chem. 282, 8777-8785 (2007)]. Thus, the kanR casette CANNOT be removed because there are NO FRT sites flanking kanR.
In the presence of 0.4-1 mM Tyr, tyrB is repressed and Leu is required for growth in minimal medium. This strategy can only be applicable for a short-term cultivation but not suitable for a long-term cultivation for heterologous expression of foreign genes.
Each target gene was deleted from the chromosome of C43 (DE3) E. coli strain using λ-Red recombination system.
Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
Proper citation: RRID:Addgene_61908 Copy
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21925267
Comments: Genotype = cyo::kan argH (Kan cassette in the chromosomal DNA)
Precursor strain = CLY (Genotype cyo::kan)
Selective amino acid labeling (and/or requirement) = Arg
CLY strain was derived from the C43(DE3)strain by transferring the cyo deletion (with kanamycin resistance cassette kanR) from an E. coli B strain to C43(DE3) strain by phage P1 [J. Biol. Chem. 282, 8777-8785 (2007)]. Thus, the kanR casette CANNOT be removed because there are NO FRT sites flanking kanR.
Each target gene was deleted from the chromosome of C43 (DE3) E. coli strain using λ-Red recombination system.
Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
Proper citation: RRID:Addgene_61909 Copy
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21925267
Comments: Genotype = cyo::kan ilvE avtA aspC (Kan cassette in the chromosomal DNA)
Precursor strain = ML6 (Genotype cyo::kan)
Selective amino acid labeling (and/or requirement) = (Ala#), Ile, Leu*, Tyr#, Val
CLY strain was derived from the C43(DE3)strain by transferring the cyo deletion (with kanamycin resistance cassette kanR) from an E. coli B strain to C43(DE3) strain by phage P1 [J. Biol. Chem. 282, 8777-8785 (2007)]. Thus, the kanR casette CANNOT be removed because there are NO FRT sites flanking kanR.
*In the presence of 0.4-1 mM Tyr, tyrB is repressed and Leu is required for growth in minimal medium. This strategy can only be applicable for a short-term cultivation but not suitable for a long-term cultivation for heterologous expression of foreign genes [Methods 55, 370-378 (2011)].
#In the presence of 0.4-1 mM Tyr, tyrB is repressed and Tyr is required for growth in minimal medium. Under these conditions, this strain can also be used for selective labeling of the input Tyr and/or Ala label(s), although minor diffusion of the input label(s) can occur (see Tables 2, 3). Note that this strategy can only be applicable for a short-term cultivation but not suitable for a long-term cultivation for heterologous expression of foreign geness [Methods 55, 370-378 (2011)].
Each target gene was deleted from the chromosome of C43 (DE3) E. coli strain using λ-Red recombination system.
Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
Proper citation: RRID:Addgene_61910 Copy
Species: Homo sapiens
Genetic Insert: Rab4B
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4700; Vector Backbone:pAcGFP1-C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:25416943
Proper citation: RRID:Addgene_61801 Copy
Species: Homo sapiens
Genetic Insert: Rtn4a
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4700; Vector Backbone:pAcGFP1-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:18442980
Comments: This plasmid has been found to run as a multimer. Plasmid multimerization often does not impact plasmid function, but may reduce transformation efficiencies. You may need to screen multiple colonies to isolate the monomeric version of this plasmid. If you still have trouble isolating the monomeric version, you might consider linearizing, gel extracting, re-ligating, and transforming the plasmid.
Proper citation: RRID:Addgene_61807 Copy
Species: Homo sapiens
Genetic Insert: UBL4A
Vector Backbone Description: Backbone Marker:Origene; Backbone Size:4929; Vector Backbone:pCMV6-ENTRY; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:24424410
Proper citation: RRID:Addgene_61809 Copy
Species: Homo sapiens
Genetic Insert: AICDA
Vector Backbone Description: Backbone Marker:Clontech; Vector Backbone:pEGFP-C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:24324754
Proper citation: RRID:Addgene_61926 Copy
Species: Thermoplasma volcanium
Genetic Insert: TvoVMAN11 intein
Vector Backbone Description: Backbone Marker:Novagen; Backbone Size:3559; Vector Backbone:pRSFDuet; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:24574026
Proper citation: RRID:Addgene_61813 Copy
Species: Pyrococcus horikoshii
Genetic Insert: PhoRadAdC6 intein
Vector Backbone Description: Backbone Marker:Novagen; Backbone Size:3559; Vector Backbone:pRSFDuet; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:24574026
Proper citation: RRID:Addgene_61817 Copy
Species: Homo sapiens
Genetic Insert: cyclin A
Vector Backbone Description: Backbone Marker:Pines Lab; Vector Backbone:pt7 venus cks1 (1843); Vector Types:in vitro transcription; Bacterial Resistance:Kanamycin
Defining Citation: PMID:20733051
Proper citation: RRID:Addgene_61839 Copy
Species: Homo sapiens
Genetic Insert: cyclin A
Vector Backbone Description: Backbone Marker:unknown; Vector Backbone:pCeru-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Proper citation: RRID:Addgene_61842 Copy
Species: Homo sapiens
Genetic Insert: Tmem214
Vector Backbone Description: Backbone Marker:Clontech; Vector Backbone:pEGFP-N2; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:20876400
Proper citation: RRID:Addgene_62026 Copy
Species: E. coli
Genetic Insert: lacZ alpha
Vector Backbone Description: Vector Backbone:pEGFP; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Proper citation: RRID:Addgene_61972 Copy
Species: Homo sapiens
Genetic Insert: Tmem194
Vector Backbone Description: Backbone Marker:Clontech; Vector Backbone:pEGFP-N2; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:20876400
Proper citation: RRID:Addgene_62027 Copy
Species: Homo sapiens
Genetic Insert: NET33
Vector Backbone Description: Backbone Marker:unknown; Vector Backbone:pmRFP-N2; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:20091084
Proper citation: RRID:Addgene_61979 Copy
Species: Homo sapiens
Genetic Insert: Tmem41A
Vector Backbone Description: Backbone Marker:unknown; Vector Backbone:pEGPF-N2; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:20693407
Proper citation: RRID:Addgene_62023 Copy
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