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http://www.addgene.org/102672

Species: Synthetic
Genetic Insert: GFPmut2
Vector Backbone Description: Backbone Marker:Genewiz; Vector Backbone:pUC57 (ori changed to p15A); Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29084263
Comments: Additional Insert: TetR . Please visit https://www.biorxiv.org/content/early/2017/09/23/192963 for bioRxiv preprint.

Proper citation: RRID:Addgene_102672 Copy   

•   Source: Addgene

http://www.addgene.org/102790

Species: T7 phage
Genetic Insert: T7 RNA polymerase
Vector Backbone Description: Backbone Marker:Qiagen; Vector Backbone:pQE; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:29120463

Proper citation: RRID:Addgene_102790 Copy   

•   Source: Addgene

http://www.addgene.org/102799

Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
Defining Citation: PMID:33289521
Comments: This work is supported in part by JSPS-NSF International Collaborations in Chemistry (ICC) research grant. Genotype= aspC tyrB trpA trpB glyA serB Precursor strain = RF14 Modified from the parent Escherichia coli BL21(DE3) strain Selective amino acid labeling (and/or requirement) = Asp, Tyr, Trp, (Phe), Gly, Ser+++++ +++++ RF15 has knockouts in aspC, tyrB, trpA, trpB, glyA and serB genes and requires the presence of L-Asp, L-Tyr, L-Trp, L-Gly plus L-Ser for growth in M63 minimal medium, but it does NOT grow in the presence of L-Asp, L-Tyr, L-Trp, L-Gly, L-Ser plus L-Cys (either in the presence or absence of L-Ala) (i.e., L-Cys inhibits the growth of RF15) Please visit the following links for additional details on this strain and selective amino acid labeling- http://www2.nms.ac.jp/fesworld/EcoliStrains.html http://www2.nms.ac.jp/fesworld/EcoliStrainsSuppl.html Note that these strains are NOT competent cells and one needs to make them competent before use. 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. Supporting References: Lin, M. T., Fukazawa, R., Miyajima-Nakano, Y., Matsushita, S., Choi, S. K., Iwasaki, T., and Gennis, R. B. (2015) Escherichia coliauxotroph host strains for amino acid-selective isotope labeling of recombinant proteins. Methods Enzymol. (Isotope Labeling of Biomolecules - Labeling Methods), 565, 45-66. Iwasaki, T., Fukazawa, R., Miyajima-Nakano, Y., Baldansuren, A., Matsushita, S., Lin, M. T., Gennis, R. B., Hasegawa, K., Kumasaka, T., and Dikanov, S. A. (2012) Dissection of hydrogen bond interaction network around an iron-sulfur cluster by site-specific isotope labeling of hyperthermophilic archaeal Rieske-type ferredoxin. J. Am. Chem. Soc. 134, 19731-19738. Lin, M. T., Sperling, L. J., Frericks Schmidt, H. L., Tang, M., Samoilova, R. I., Kumasaka, T., Iwasaki, T., Dikanov, S. A., Rienstra, C. M., and Gennis, R. B. (2011) A rapid and robust method for selective isotope labeling of proteins. Methods 55, 370-378.

Proper citation: RRID:Addgene_102799 Copy   

•   Source: Addgene

http://www.addgene.org/102714

Genetic Insert: Linker sequence
Vector Backbone Description: Backbone Marker:Self-made; Backbone Size:2267; Vector Backbone:derived from pSB1K3; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:29293531

Proper citation: RRID:Addgene_102714 Copy   

•   Source: Addgene

http://www.addgene.org/102674

Species: Synthetic
Genetic Insert: GFPmut2
Vector Backbone Description: Backbone Marker:Genewiz; Vector Backbone:pUC57 (ori changed to p15A); Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29084263
Comments: Additional Insert: TetR . Please visit https://www.biorxiv.org/content/early/2017/09/23/192963 for bioRxiv preprint.

Proper citation: RRID:Addgene_102674 Copy   

•   Source: Addgene

http://www.addgene.org/102677

Species: Homo sapiens
Genetic Insert: RIPK1
Vector Backbone Description: Backbone Size:7166; Vector Backbone:pMIG; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26195820
Comments: The RIPK1 insert has a K87Q polymorphism which has no effect on protein activity.

Proper citation: RRID:Addgene_102677 Copy   

•   Source: Addgene

http://www.addgene.org/102710

Genetic Insert: Linker sequence
Vector Backbone Description: Backbone Marker:Self-made; Backbone Size:2267; Vector Backbone:derived from pSB1K3; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:29293531

Proper citation: RRID:Addgene_102710 Copy   

•   Source: Addgene

http://www.addgene.org/102808

Genetic Insert: HP1cs-Frb1x
Vector Backbone Description: Vector Backbone:pLenti; Vector Types:Mammalian Expression, Lentiviral, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:28916764

Proper citation: RRID:Addgene_102808 Copy   

•   Source: Addgene

http://www.addgene.org/102809

Genetic Insert: HP1cs-Frb2x
Vector Backbone Description: Vector Backbone:pLenti; Vector Types:Mammalian Expression, Lentiviral, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:28916764

Proper citation: RRID:Addgene_102809 Copy   

•   Source: Addgene

http://www.addgene.org/102803

Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
Defining Citation: PMID:33289521
Comments: This work is supported in part by JSPS-NSF International Collaborations in Chemistry (ICC) research grant. Genotype= aspC tyrB ilvE avtA yfbQ(alaA) yfdZ(alaC) Precursor strain = RF18 Modified from the parent Escherichia coli BL21(DE3) strain Selective amino acid labeling (and/or requirement) = Asp, Tyr, Phe, Ile, Leu, Val#### #### RF21 has knockouts in the four general transaminase genes of E. coli (aspC, tyrB, ilvE, and avtA) and is found to require the presence of L-Asp, L-Tyr, L-Phe, L-Ile, L-Leu plus L-Val for slow growth in M63 minimal medium. Although RF21 strain has further knockouts in yfbQ (alaA) and yfdZ (alaC) genes, it is NOT an L-Ala auxotroph, either (requiring the presence of L-Asp, L-Tyr, L-Phe, L-Ile, L-Leu plus L-Val for slow growth in M63 minimal medium, like RF18). Please visit the following links for additional details on this strain and selective amino acid labeling- http://www2.nms.ac.jp/fesworld/EcoliStrains.html http://www2.nms.ac.jp/fesworld/EcoliStrainsSuppl.html Note that these strains are NOT competent cells and one needs to make them competent before use. 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. Supporting References: Lin, M. T., Fukazawa, R., Miyajima-Nakano, Y., Matsushita, S., Choi, S. K., Iwasaki, T., and Gennis, R. B. (2015) Escherichia coliauxotroph host strains for amino acid-selective isotope labeling of recombinant proteins. Methods Enzymol. (Isotope Labeling of Biomolecules - Labeling Methods), 565, 45-66. Iwasaki, T., Fukazawa, R., Miyajima-Nakano, Y., Baldansuren, A., Matsushita, S., Lin, M. T., Gennis, R. B., Hasegawa, K., Kumasaka, T., and Dikanov, S. A. (2012) Dissection of hydrogen bond interaction network around an iron-sulfur cluster by site-specific isotope labeling of hyperthermophilic archaeal Rieske-type ferredoxin. J. Am. Chem. Soc. 134, 19731-19738. Lin, M. T., Sperling, L. J., Frericks Schmidt, H. L., Tang, M., Samoilova, R. I., Kumasaka, T., Iwasaki, T., Dikanov, S. A., Rienstra, C. M., and Gennis, R. B. (2011) A rapid and robust method for selective isotope labeling of proteins. Methods 55, 370-378.

Proper citation: RRID:Addgene_102803 Copy   

•   Source: Addgene

http://www.addgene.org/102806

Genetic Insert: MS2-Fkbp
Vector Backbone Description: Vector Backbone:pLenti; Vector Types:Mammalian Expression, Lentiviral, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:28916764

Proper citation: RRID:Addgene_102806 Copy   

•   Source: Addgene

http://www.addgene.org/102646

Vector Backbone Description: Backbone Marker:Markus Ralser (Addgene plasmid # 19407), Clontech = pTRE-T2; Vector Backbone:pTRE-Tight2; Vector Types:Mammalian Expression, RNAi; Bacterial Resistance:Ampicillin
Comments: In fusion cloning was used to insert a synthetic DNA template containing the mIR cassette from the pCDNA 6.2-GW/miR into the EcoRI and XbaI site of pTRE-T2-Tight. I engineered two BsaI sites into the miR cassette so users can digest the vector with BsaI for directional cloning of Blockit microRNAs using the Block-IT RNAi designer engine from Invitrogen. To make the BsaI sites unique I mutated out Bsa I sites in the original pTRE-T2 Tight vector and also removed XhoI and XbaI sites by site-directed mutagenesis from the pTRE-T2 vector and the corresponding puromycin/hygromycin resistance genes. I also dropped in a unique NheI and EagI restriction site between the ampicillin gene and the pMB101 origin of replication sequence so that the puromycin and hygromin genes could be placed into the pTRE-T2-miR vector with the infusion cloning method. Investigators can remove the miR cassette from the pTRE-T2 puromycin/hygromycin vector and using cloning methods to insert any cDNA of interest into the site for doxycline-inducible expression of the target cDNA. A single vector can be utilized from the production of tetracycline inducible miRNAs or cDNAs. For puromycin use .25 micrograms/ml to .50 micrograms/ml to get clones after a couple of weeks. Growth response curve tested in 293HEK cells. Suggest methylation deficient cells from NEB to alleviate methylation sensitive restriction sites like Xba I. However not recommended for long-term propagation and storage.

Proper citation: RRID:Addgene_102646 Copy   

•   Source: Addgene

http://www.addgene.org/102802

Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
Defining Citation: PMID:33289521
Comments: This work is supported in part by JSPS-NSF International Collaborations in Chemistry (ICC) research grant. Genotype= aspC tyrB ilvE avtA Precursor strain = RF17 Modified from the parent Escherichia coli BL21(DE3) strain Selective amino acid labeling (and/or requirement) = Asp, Tyr, Phe, Ile, Leu, Val#### ####RF18 has knockouts in the four general transaminase genes of E. coli (aspC, tyrB, ilvE, and avtA) and is found to require the presence of L-Asp, L-Tyr, L-Phe, L-Ile, L-Leu plus L-Val for slow growth in M63 minimal medium. Please visit the following links for additional details on this strain and selective amino acid labeling- http://www2.nms.ac.jp/fesworld/EcoliStrains.html http://www2.nms.ac.jp/fesworld/EcoliStrainsSuppl.html Note that these strains are NOT competent cells and one needs to make them competent before use. 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. Supporting References: Lin, M. T., Fukazawa, R., Miyajima-Nakano, Y., Matsushita, S., Choi, S. K., Iwasaki, T., and Gennis, R. B. (2015) Escherichia coliauxotroph host strains for amino acid-selective isotope labeling of recombinant proteins. Methods Enzymol. (Isotope Labeling of Biomolecules - Labeling Methods), 565, 45-66. Iwasaki, T., Fukazawa, R., Miyajima-Nakano, Y., Baldansuren, A., Matsushita, S., Lin, M. T., Gennis, R. B., Hasegawa, K., Kumasaka, T., and Dikanov, S. A. (2012) Dissection of hydrogen bond interaction network around an iron-sulfur cluster by site-specific isotope labeling of hyperthermophilic archaeal Rieske-type ferredoxin. J. Am. Chem. Soc. 134, 19731-19738. Lin, M. T., Sperling, L. J., Frericks Schmidt, H. L., Tang, M., Samoilova, R. I., Kumasaka, T., Iwasaki, T., Dikanov, S. A., Rienstra, C. M., and Gennis, R. B. (2011) A rapid and robust method for selective isotope labeling of proteins. Methods 55, 370-378.

Proper citation: RRID:Addgene_102802 Copy   

•   Source: Addgene

http://www.addgene.org/102643

Vector Backbone Description: Backbone Marker:Invitrogen; Vector Backbone:pcDNA3_neo; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Comments: No insert, but contains 3 HA epitopes for c-terminal tagged proteins

Proper citation: RRID:Addgene_102643 Copy   

•   Source: Addgene

http://www.addgene.org/102644

Vector Backbone Description: Backbone Marker:Invitrogen; Vector Backbone:pcDNA3_neo; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:27365400
Comments: No insert, but contains a single N-terminus Strepavidin binding peptide and Flag epitopes

Proper citation: RRID:Addgene_102644 Copy   

•   Source: Addgene

http://www.addgene.org/102770

Species: Other
Genetic Insert: ChrimsonR::FusionRed::Kv2.1
Vector Backbone Description: Backbone Size:5366; Vector Backbone:pAAV; Vector Types:AAV; Bacterial Resistance:Ampicillin
Defining Citation: PMID:33495637

Proper citation: RRID:Addgene_102770 Copy   

•   Source: Addgene

http://www.addgene.org/102771

Species: Other
Genetic Insert: C1V1::FusionRed::Kv2.1
Vector Backbone Description: Backbone Size:5366; Vector Backbone:pAAV; Vector Types:AAV; Bacterial Resistance:Ampicillin

Proper citation: RRID:Addgene_102771 Copy   

•   Source: Addgene

http://www.addgene.org/102810

Genetic Insert: MS2-HP1cs
Vector Backbone Description: Vector Backbone:pLenti; Vector Types:Mammalian Expression, Lentiviral, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:28916764

Proper citation: RRID:Addgene_102810 Copy   

•   Source: Addgene

http://www.addgene.org/102748

Species: Homo sapiens
Genetic Insert: EPHA3
Vector Backbone Description: Backbone Marker:Invitrogen; Vector Backbone:pcDNA3.1(+)/myc-His B; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22829656
Comments: Please note last 10 amino acids in C-terminal are omitted in these constructs. These series of EPHA3 constructs carry missense mutations found in lung and colon cancer.

Proper citation: RRID:Addgene_102748 Copy   

•   Source: Addgene

http://www.addgene.org/102745

Species: Homo sapiens
Genetic Insert: EPHA3
Vector Backbone Description: Backbone Marker:Invitrogen; Vector Backbone:pcDNA3.1(+)/myc-His B; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22829656
Comments: Please note last 10 amino acids in C-terminal are omitted in these constructs. These series of EPHA3 constructs carry missense mutations found in lung and colon cancer.

Proper citation: RRID:Addgene_102745 Copy   

•   Source: Addgene