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Species: Homo sapiens
Genetic Insert: gRNA targeting human cGAS
Vector Backbone Description: Vector Backbone:pLCRISPR; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26229115
Proper citation: RRID:Addgene_102610 Copy
Species: Chimera
Genetic Insert: KcsA-Kv1.3
Vector Backbone Description: Backbone Size:2546; Vector Backbone:pBEST; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: Please visit https://www.biorxiv.org/content/early/2017/01/30/104455 for bioRxiv preprint.
Proper citation: RRID:Addgene_102457 Copy
Vector Backbone Description: Vector Backbone:pTRIP; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26229115
Proper citation: RRID:Addgene_102611 Copy
Species: Other
Genetic Insert: EGFP
Vector Backbone Description: Backbone Marker:MPI-CBG PEP facility; Vector Backbone:pOEM1; Vector Types:Mammalian Expression, Insect Expression, Baculoviral rescue shuttle vector; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29198566
Proper citation: RRID:Addgene_102782 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: Please visit https://www.biorxiv.org/content/early/2017/09/23/192963 for bioRxiv preprint.
Proper citation: RRID:Addgene_102667 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: tryptophanyl tRNA synthetase
Vector Backbone Description: Vector Backbone:pRST11B; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29120463
Proper citation: RRID:Addgene_102789 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: Please visit https://www.biorxiv.org/content/early/2017/09/23/192963 for bioRxiv preprint.
Proper citation: RRID:Addgene_102669 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: 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: 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: 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: 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
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
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
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
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