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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

http://www.addgene.org/102740

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_102740 Copy   

•   Source: Addgene

http://www.addgene.org/102741

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_102741 Copy   

•   Source: Addgene

http://www.addgene.org/102742

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_102742 Copy   

•   Source: Addgene

http://www.addgene.org/102759

Species: Mus musculus
Genetic Insert: p27
Vector Backbone Description: Vector Backbone:lentiCRISPR v2; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:28581527

Proper citation: RRID:Addgene_102759 Copy   

•   Source: Addgene

http://www.addgene.org/102750

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_102750 Copy   

•   Source: Addgene

http://www.addgene.org/102755

Species: Homo sapiens
Genetic Insert: EphA2
Vector Backbone Description: Vector Backbone:pCLXSN; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24607842

Proper citation: RRID:Addgene_102755 Copy   

•   Source: Addgene

http://www.addgene.org/102758

Species: Homo sapiens
Genetic Insert: c-Jun
Vector Backbone Description: Vector Backbone:pCLXSN; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24607842

Proper citation: RRID:Addgene_102758 Copy   

•   Source: Addgene

http://www.addgene.org/102752

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_102752 Copy   

•   Source: Addgene

http://www.addgene.org/102754

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_102754 Copy   

•   Source: Addgene

http://www.addgene.org/102680

Genetic Insert: J23106:B0034:amilCP:rrnBT1-T7TE
Vector Backbone Description: Backbone Marker:BioBricks Foundation; Backbone Size:2039; Vector Backbone:pSB1C3; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:29293531

Proper citation: RRID:Addgene_102680 Copy   

•   Source: Addgene

http://www.addgene.org/102724

Species: Homo sapiens
Genetic Insert: EFNA1
Vector Backbone Description: Vector Backbone:pAd Easy1; Vector Types:Adenoviral; Bacterial Resistance:Kanamycin
Defining Citation: PMID:17079451
Comments: This construct is a adenoviral vector which carries human EFNA1 cDNA and ready to produce adenovirus which express Ephrin-A1 by tranfection into 293A cells followed by PacI digestion.

Proper citation: RRID:Addgene_102724 Copy   

•   Source: Addgene