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Species: Saccharomyces cerevisiae
Genetic Insert: dropout cassette containing toxic genes ccdB and TPK2
Vector Backbone Description: Vector Backbone:pYTK001; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:34860007
Comments: Constructed using the Yeast Modular Cloning Toolkit (Lee et al. ACS Synth Biol. 2015).
Proper citation: RRID:Addgene_176179 Copy
Species: Synthetic
Genetic Insert: SR6v15
Vector Backbone Description: Backbone Size:5639; Vector Backbone:pETx; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:34535642
Proper citation: RRID:Addgene_176212 Copy
Species: Synthetic
Genetic Insert: 41 bp non-coding DNA spacer from pYTK048
Vector Backbone Description: Vector Backbone:pYTK001; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:34860007
Comments: Constructed using the Yeast Modular Cloning Toolkit (Lee et al. ACS Synth Biol. 2015).
Proper citation: RRID:Addgene_176178 Copy
Species: Synthetic
Genetic Insert: SR6v15.t
Vector Backbone Description: Backbone Size:5639; Vector Backbone:pETx; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:34535642
Proper citation: RRID:Addgene_176214 Copy
Species: Synthetic
Genetic Insert: SR6v15.d
Vector Backbone Description: Backbone Size:5639; Vector Backbone:pETx; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:34535642
Proper citation: RRID:Addgene_176213 Copy
Species: Other
Genetic Insert: GFP
Vector Backbone Description: Backbone Size:5050; Vector Backbone:pAAV-CBA-w; Vector Types:Mammalian Expression, AAV; Bacterial Resistance:Ampicillin
Defining Citation: PMID:32184391
Proper citation: RRID:Addgene_176215 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: 5' homology for integration site XI-3
Vector Backbone Description: Vector Backbone:pYTK001; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:34860007
Comments: Constructed using the Yeast Modular Cloning Toolkit (Lee et al. ACS Synth Biol. 2015). Homologous region amplified from EasyClone-MarkerFree toolkit (Jessop-Fabre et al. Biotechnol J. 2016).
Proper citation: RRID:Addgene_176170 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: 5' homology for integration site XI-5
Vector Backbone Description: Vector Backbone:pYTK001; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:34860007
Comments: Constructed using the Yeast Modular Cloning Toolkit (Lee et al. ACS Synth Biol. 2015). Homologous region amplified from EasyClone-MarkerFree toolkit (Jessop-Fabre et al. Biotechnol J. 2016).
Proper citation: RRID:Addgene_176172 Copy
Species: Homo sapiens
Genetic Insert: HLA-DRa promoter
Vector Backbone Description: Backbone Marker:Dr. Didier Trono; Backbone Size:0; Vector Backbone:SIN LV pRLLhPGK.GFP Sin-18 (modified); Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:11781219
Comments: SIN lentiviral vector that expresses GFP controlled by the promoter of the human HLA-DRalpha gene, which is selectively expressed in antigen-presenting cells (APCs).
We constructed DR.GFP by using the human (MHC II) HLA-DRalpha promoter (nucleotide 182 to 480, as numbered in GenBank accession X00274) to replace the PGK promoter (XhoI to BamHI sites in the PGK.GFP vector). This 300-base pair promoter sequence has been shown to confer selective expression of a transgene in MHC II+ cells.
Recombinant LVs were generated using the 3-plasmid system by cotransfection of 293T cells through calcium phosphate precipitation. The ratio of a transducing vector, pMD.G, and pCMVDeltaR8.91 was fixed at 1.5:0.5:2 µg for 10^6 293T cells.
Proper citation: RRID:Addgene_17617 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: 3' homology for integration site XII-2
Vector Backbone Description: Vector Backbone:pYTK001; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:34860007
Comments: Constructed using the Yeast Modular Cloning Toolkit (Lee et al. ACS Synth Biol. 2015). Homologous region amplified from EasyClone-MarkerFree toolkit (Jessop-Fabre et al. Biotechnol J. 2016).
Proper citation: RRID:Addgene_176175 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: 5' homology for integration site XII-2
Vector Backbone Description: Vector Backbone:pYTK001; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:34860007
Comments: Constructed using the Yeast Modular Cloning Toolkit (Lee et al. ACS Synth Biol. 2015). Homologous region amplified from EasyClone-MarkerFree toolkit (Jessop-Fabre et al. Biotechnol J. 2016).
Proper citation: RRID:Addgene_176174 Copy
Species: Homo sapiens
Genetic Insert: mCherry-INBoxAAA-eDHFR
Vector Backbone Description: Vector Backbone:pEM791; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:33904910
Proper citation: RRID:Addgene_176207 Copy
Species: Synthetic
Genetic Insert: VHH template2-3XMyc-Spacer
Vector Backbone Description: Backbone Size:2561; Vector Backbone:pBx1; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:34535642
Proper citation: RRID:Addgene_176209 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: 5' homology for integration site XI-1
Vector Backbone Description: Vector Backbone:pYTK001; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:34860007
Comments: Constructed using the Yeast Modular Cloning Toolkit (Lee et al. ACS Synth Biol. 2015). Homologous region amplified from EasyClone-MarkerFree toolkit (Jessop-Fabre et al. Biotechnol J. 2016).
Proper citation: RRID:Addgene_176166 Copy
Species: E. coli
Genetic Insert: mCherry-eDHFR
Vector Backbone Description: Backbone Marker:Dr. Eugene Makeyev; Backbone Size:-1; Vector Backbone:pEM791; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:33904910
Comments: Please note that there are minor differences between the Addgene NGS result and the depositor's sequence. These differences do not alter plasmid function.
Proper citation: RRID:Addgene_176201 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: 3' homology for integration site XI-1
Vector Backbone Description: Vector Backbone:pYTK001; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:34860007
Comments: Constructed using the Yeast Modular Cloning Toolkit (Lee et al. ACS Synth Biol. 2015). Homologous region amplified from EasyClone-MarkerFree toolkit (Jessop-Fabre et al. Biotechnol J. 2016).
Proper citation: RRID:Addgene_176167 Copy
Species: E. coli E2348/69 O127:H6
Genetic Insert: gfcD
Vector Backbone Description: Backbone Marker:from Dr Frank Collart; Vector Backbone:pBH31; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:34780538
Comments: encodes (pelC signal sequence)-His10-GfcD(22-698)
Proper citation: RRID:Addgene_176200 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: 3' homology for integration site XI-2
Vector Backbone Description: Vector Backbone:pYTK001; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:34860007
Comments: Constructed using the Yeast Modular Cloning Toolkit (Lee et al. ACS Synth Biol. 2015). Homologous region amplified from EasyClone-MarkerFree toolkit (Jessop-Fabre et al. Biotechnol J. 2016).
Proper citation: RRID:Addgene_176169 Copy
Species: Homo sapiens
Genetic Insert: mCherry-INBox-eDHFR
Vector Backbone Description: Vector Backbone:pEM791; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:33904910
Proper citation: RRID:Addgene_176202 Copy
Species: Mus musculus
Genetic Insert: delFasL
Vector Backbone Description: Backbone Size:0; Vector Backbone:EF.CMV.GFP; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:12788657
Comments: The CMV promoter was inserted immediately upstream of GFP in the EF.GFP LV to make the intermediate LV, EF.V-CMV.GFP, with a unique EcoRV site available to insert a gene of interest under the control of the EF promoter. Noncleavable mouse delFasL cDNA was constructed using PCR cloning to delete the exon 2 region, which contains the metalloproteinase cleavage site. The delFasL gene was then cloned into the EcoRV site of EF.CMV.GFP to make the EF.delFasL-CMV.GFP LV.
Recombinant LVs were produced by transient transfection of the transducing vector into 293T cells with two packaging vectors: pMD.G, a plasmid expressing the VSV-G envelope gene, and pCMVDeltaR8.91, a plasmid expressing the HIV-1 gag/pol, tat, and rev genes.
Proper citation: RRID:Addgene_17620 Copy
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