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Species: Homo sapiens
Genetic Insert: FLAG-HaloTag-TERT Exon 1 and part of Exon 2
Vector Backbone Description: Vector Backbone:pFastBac; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:27523609
Proper citation: RRID:Addgene_86843 Copy
Vector Backbone Description: Backbone Size:7267; Vector Backbone:pET; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Comments: This plasmid is an empty vector. Your gene can be inserted with a LIC cloning protocol. This plasmid can be used as a single-expression vector, and it is also compatible with our 2-series polycistronic destination vectors (2D, 2E, and 2Z), if co-expression with other genes is desired.
To clone into this vector, add LIC tags to the 5' end of your PCR primers.
Forward - 5' TACTTCCAATCCAATGCA 3'
Reverse - 5' CTCCCACTACCAATGCC 3'
Do NOT include a stop codon with your reverse primer.
Linearize the plasmid with SspI, then gel purify.
When digesting the DNA with T4 polymerase, use dGTP for the insert and dCTP for your linearized vector.
More information on this vector can be found through http://qb3.berkeley.edu/qb3/macrolab/
Proper citation: RRID:Addgene_86933 Copy
Species: Mus musculus
Genetic Insert: postLib-mRFP-4xboxB
Vector Backbone Description: Vector Backbone:unknown; Vector Types:Mammalian Expression, Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:28449067
Proper citation: RRID:Addgene_87006 Copy
Species: Mus musculus
Genetic Insert: preCLAP
Vector Backbone Description: Vector Backbone:unknown; Vector Types:Mammalian Expression, Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:28449067
Proper citation: RRID:Addgene_87003 Copy
Species: Mus musculus
Genetic Insert: postCLAP
Vector Backbone Description: Vector Backbone:unknown; Vector Types:Mammalian Expression, Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:28449067
Proper citation: RRID:Addgene_87004 Copy
Species: Staphylococcus aureus
Genetic Insert: S.aureus SrtA 5M
Vector Backbone Description: Backbone Size:5400; Vector Backbone:pET30b; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:28088451
Comments: Pentamutant sortase A was originally described by David R. Liu's laboratory (Harvard University) in: A General Strategy for the Evolution of Bond-Forming Enzymes Using Yeast Display. Chen I, Dorr BM, Liu DR. Proc Natl Acad Sci U S A. 2011 Jul 12;108(28):11399-404 (PubMed ID: 21697512). As noted in this publication, the cloned sortase A gene lacks the N-terminal 59 amino acids, which do not impact catalytic activity, but these amino acids are still included in the numbering for the mutations.
Proper citation: RRID:Addgene_86962 Copy
Species: Yersinia enterocolitica
Genetic Insert: irp3 WT
Vector Backbone Description: Backbone Marker:novagen; Backbone Size:5368; Vector Backbone:pET28b; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:23066849
Comments: Biochemistry 55 (2016) 5423-5433
Proper citation: RRID:Addgene_87106 Copy
Species: Pseudomonas aeruginosa PAO1
Genetic Insert: pchG WT
Vector Backbone Description: Backbone Marker:novagen; Backbone Size:5368; Vector Backbone:pET28b; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:27601130
Proper citation: RRID:Addgene_87107 Copy
Species: Pseudomonas aeruginosa PAO1
Genetic Insert: pchA WT
Vector Backbone Description: Backbone Marker:novagen; Backbone Size:5368; Vector Backbone:pET28b; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:23942051
Comments: Archives of Biochemistry and Biophysics, 539 (2013) 70-80, PMC3836504
JACS 138 (2016) 9277-9293, PMC 5029964
Proper citation: RRID:Addgene_87102 Copy
Species: Synthetic
Genetic Insert: NADPH Alanine Dehydrogenase
Vector Backbone Description: Backbone Marker:Lucigen (Middleton, WI); Backbone Size:1800; Vector Backbone:pSMART-HCKan (AF532107.1); Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:34600151
Comments: Read the preprint at bioRxiv: https://www.biorxiv.org/content/10.1101/2020.08.30.274290v1.full.
Proper citation: RRID:Addgene_87172 Copy
Species: Caenorhabditis elegans
Genetic Insert: smo-1
Vector Backbone Description: Backbone Size:5365; Vector Backbone:pHISTEV30a; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:25475837
Proper citation: RRID:Addgene_87054 Copy
Species: Caenorhabditis elegans
Genetic Insert: UBC-9
Vector Backbone Description: Backbone Size:5365; Vector Backbone:PpHISTEV30a; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:25475837
Proper citation: RRID:Addgene_87055 Copy
Species: Homo sapiens
Genetic Insert: Spinophilin
Vector Backbone Description: Vector Backbone:pDONR221; Vector Types:Donor vector; Bacterial Resistance:Kanamycin
Defining Citation: PMID:28634551
Proper citation: RRID:Addgene_87123 Copy
Species: Homo sapiens
Genetic Insert: TZAP
Vector Backbone Description: Vector Backbone:sv272; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:28082411
Proper citation: RRID:Addgene_87185 Copy
Species: Homo sapiens
Genetic Insert: NUP133
Vector Backbone Description: Backbone Marker:Clontech; Vector Backbone:pEGFP-C1-KSENB; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:27613095
Proper citation: RRID:Addgene_87339 Copy
Species: Homo sapiens
Genetic Insert: NUP88
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4761; Vector Backbone:pEGFP-C1X; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:27613095
Proper citation: RRID:Addgene_87338 Copy
Species: Homo sapiens
Genetic Insert: NUP93
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4761; Vector Backbone:pEGFP-C1X; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:27613095
Proper citation: RRID:Addgene_87336 Copy
Species: Homo sapiens
Genetic Insert: TOMM20 Homology Arms with linker-mEGFP
Vector Backbone Description: Vector Backbone:pUC57; Vector Types:Mammalian Expression, CRISPR, Donor Template; Bacterial Resistance:Kanamycin
Defining Citation: PMID:28814507
Comments: This plasmid has been used with locus-specific CRISPR/Cas9 to add a mEGFP tag to the C-terminus of human TOMM20 in WTC human induced pluripotent stem cells by the Allen Institute for Cell Science. Linker (AA) sequence: GGSGDPPVAT. After protein tagging using this donor template plasmid and CRISPR/Cas9 reagents, transfected cells may exhibit varying intensity levels of fluorescence, likely due to editing precision. To obtain cells of uniform intensity levels, see our protocol for fluorescence-assisted cell sorting and subcloning of transfected cells (https://www.allencell.org/instructional-videos-and-tutorials-for-cell-methods.html). Further, we recommend PCR-based assays for identifying precisely edited clones as previously described (https://www.molbiolcell.org/doi/abs/10.1091/mbc.e17-03-0209). For more information on the entire plasmid collection, please see https://www.addgene.org/allen-institute-cell-science/ .
Please visit https://www.biorxiv.org/content/early/2017/03/31/123042 for bioRxiv preprint.
Proper citation: RRID:Addgene_87423 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: CUS2
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:7300; Vector Backbone:pGBKT7; Vector Types:Bacterial Expression, Yeast Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:28062854
Proper citation: RRID:Addgene_87558 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: MUD2
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:7300; Vector Backbone:pGBKT7; Vector Types:Bacterial Expression, Yeast Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:28062854
Proper citation: RRID:Addgene_87588 Copy
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