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Genetic Insert: J23106:B0034:spisPink:rrnBT1-T7TE
Vector Backbone Description: Backbone Marker:Self-made; Backbone Size:2195; Vector Backbone:derived from pSB1K3; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:29293531
Proper citation: RRID:Addgene_102703 Copy
Species: Homo sapiens
Genetic Insert: APPL1
Vector Backbone Description: Backbone Size:4722; Vector Backbone:mCherry-C3; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Comments: *Note: Addgene's quality control sequencing finds additional residues KPNSAVDGTAGPGSTGSR appended to the C-terminus of the APPL1 protein. These additional residues are not thought to affect protein function.
Proper citation: RRID:Addgene_102784 Copy
Species: Homo sapiens
Genetic Insert: Rab5a
Vector Backbone Description: Backbone Size:4731; Vector Backbone:EGFP-C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Comments: mCherry-C2-Rab5a encodes wild-type human Rab5a,(NM_004162) as a fusion protein with mCherry. Rab5a was inserted between the Eco R1 and Sal 1 restriction sites. mCherry-C2 encodes a monomeric variant of DsRed. The backbone plasmid is pEGFP-C1 (Clontech). The C2 version of mCherry was created by insertion of a short piece of DNA between Xho 1 and EcoR1 restriction sites. In the resulting multiple cloning site, the Sac 1 restriction site has been eliminated and the Eco R1 restriction site is in frame.
Proper citation: RRID:Addgene_102785 Copy
Species: Other
Genetic Insert: reverse transcription xenopolymerase with 3'-5' deficient exonuclease
Vector Backbone Description: Vector Backbone:modified pET21; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:27339990
Proper citation: RRID:Addgene_102786 Copy
Species: Homo sapiens
Genetic Insert: Zinc finger protein 57
Vector Backbone Description: Backbone Marker:NEB; Vector Backbone:NEB DHFR control plasmid; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29159284
Proper citation: RRID:Addgene_102716 Copy
Species: Synthetic
Genetic Insert: HaloTag-bActinCDS-bActin3'UTR-MS2V5
Vector Backbone Description: Backbone Size:6250; Vector Backbone:pUbC; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:27791158
Comments: Please note, due to the AT rich sequence within the Actin 3'UTR, there are some discrepancies between the Addgene NGS result and the provided full plasmid reference sequence.
Proper citation: RRID:Addgene_102718 Copy
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_102670 Copy
Species: Other
Genetic Insert: Green Fluorescent Protein
Vector Backbone Description: Backbone Marker:Novagen; Vector Backbone:pACYC; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:29120463
Proper citation: RRID:Addgene_102791 Copy
Species: Other
Genetic Insert: Green Fluorescent Protein
Vector Backbone Description: Backbone Marker:Novagen; Vector Backbone:pACYC; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:29120463
Proper citation: RRID:Addgene_102792 Copy
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
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
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
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
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
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
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
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
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
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
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
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