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Species: Synthetic
Genetic Insert: Zebrafish optimized Cas9 (nls-zcas9-nls from Wenbiao Chen Lab plasmid pCS2-nCas9n Addgene #47929)
Vector Backbone Description: Backbone Marker:Eric Kowarz Lab (Addgene #60511); Backbone Size:6640; Vector Backbone:pSBbi-GP; Vector Types:CRISPR, Transposon; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_165484 Copy
Species:
Genetic Insert:
Vector Backbone Description: Backbone Marker:Novagen; Vector Backbone:pTriEx-1.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments: Primers for LIC cloning: Upstream: add TTAAGAAGGAGATATACT to the 5' end of gene of interest. Downstream: add GATTGGAAGTAGAGGTTCTCTGC to the 3' end of GOI. For vector: digest with BfuAI (BspMI) before LIC-treatment. Detailed cloning method available in the paper Bruni R, Kloss B. High-throughput cloning and expression of integral membrane proteins in Escherichia coli. Curr Protoc Protein Sci. 2013 Nov 5;74:29.6.1-29.6.34.
Proper citation: RRID:Addgene_165480 Copy
Species:
Genetic Insert:
Vector Backbone Description: Backbone Marker:Novagen; Vector Backbone:pTriEx-1.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments: Primers for LIC cloning: Upstream: add TTAAGAAGGAGATATACT to the 5' end of gene of interest. Downstream: add GATTGGAAGTAGAGGTTCTCTGC to the 3' end of GOI. For vector: digest with BfuAI (BspMI) before LIC-treatment. Detailed cloning method available in the paper Bruni R, Kloss B. High-throughput cloning and expression of integral membrane proteins in Escherichia coli. Curr Protoc Protein Sci. 2013 Nov 5;74:29.6.1-29.6.34.
Proper citation: RRID:Addgene_165481 Copy
Species: Homo sapiens
Genetic Insert: Membrane bound O-acyltransferase domain containing 7
Vector Backbone Description: Backbone Marker:Stratagene /Agilent; Backbone Size:5200; Vector Backbone:pCMV-3Tag-8 modified MCS; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments: Backbone contains new MCS (multiple cloning site). CDS without stop codon was inserted with XhoI-NotI and it expresses in frame with 3xFLAG epitope tag. Construct insertion = Xhoi-GCC-[MBOAT7 CDS 1416 bp, no stop]-Noti-A-Clai-GTCGAG-3xFLAG-STOP
Proper citation: RRID:Addgene_165474 Copy
Species:
Genetic Insert:
Vector Backbone Description: Backbone Marker:Novagen; Vector Backbone:pET23; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments: Primers for LIC cloning: Upstream: add TTAAGAAGGAGATATACT to the 5' end of gene of interest. Downstream: add TGAAAATAGAGGTTTTCGGC to the 3' end of GOI. Detailed cloning method available in the paper Bruni R, Kloss B. High-throughput cloning and expression of integral membrane proteins in Escherichia coli. Curr Protoc Protein Sci. 2013 Nov 5;74:29.6.1-29.6.34.
Proper citation: RRID:Addgene_165477 Copy
Species:
Genetic Insert:
Vector Backbone Description: Backbone Marker:Novagen; Vector Backbone:pET23; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments: Primers for LIC cloning: Upstream: add TATTTTCAATCCTACGTA to the 5' end of gene of interest. Downstream: add CCCTCAATATTATACGGG to the 3' end of GOI. Detailed cloning method available in the paper Bruni R, Kloss B. High-throughput cloning and expression of integral membrane proteins in Escherichia coli. Curr Protoc Protein Sci. 2013 Nov 5;74:29.6.1-29.6.34.
Proper citation: RRID:Addgene_165478 Copy
Species: Homo sapiens
Genetic Insert: Adenosine deaminase RNA specific B2 (inactive)
Vector Backbone Description: Backbone Marker:Stratagene /Agilent; Backbone Size:5200; Vector Backbone:pCMV-3Tag-8 modified MCS; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments: Backbone contains new MCS (multiple cloning site). MCS is changed to: ctcgagctgaagcttcatggatccaaagcggccgcaatcgat (XhoI-ctg-HindIII-cat-BamHI-aaa-NotI-a-ClaI). CDS without stop codon was inserted into MCS at unkown restriction enzyme sites, and it expresses in frame with 3xFLAG epitope tag. Xhoi was probably the 5' cloning site.
Proper citation: RRID:Addgene_165470 Copy
Species: Mus musculus
Genetic Insert: Mettl3 C-terminus + linker + FKBP-V
Vector Backbone Description: Vector Backbone:None; Vector Types:Mouse Targeting; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_165421 Copy
Species: Homo sapiens
Genetic Insert: PPAR gamma
Vector Backbone Description: Backbone Marker:Amersham; Backbone Size:5000; Vector Backbone:pGEX-2TK; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments: Note that there are some discrepancies between the insert in this plasmid and the canonical GenBank PPARgamma. The Vogelstein lab maintains that this plasmid worked as specified when they made and used it.
Proper citation: RRID:Addgene_16549 Copy
Species: Homo sapiens
Genetic Insert: gRNA against PGC-1a variant 1
Vector Backbone Description: Backbone Size:9580; Vector Backbone:pSico; Vector Types:Mammalian Expression, Lentiviral, CRISPR; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_165425 Copy
Species: Homo sapiens
Genetic Insert: p73
Vector Backbone Description: Backbone Size:5210; Vector Backbone:pBI-MCS-EGFP; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments: Tet-responsive p73 expression construct.
Proper citation: RRID:Addgene_16545 Copy
Species:
Genetic Insert:
Vector Backbone Description: Backbone Size:5210; Vector Backbone:pBI-MCS-EGFP; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments: The tet-responsive reporter plasmid pBI-EGFP was purchased from Clontech and modified by the inclusion of a polylinker to create pBI-MCS-EGFP.
Proper citation: RRID:Addgene_16542 Copy
Species:
Genetic Insert: tTA
Vector Backbone Description: Backbone Size:6000; Vector Backbone:pMk10-59; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments: tTA is a a tetracycline-controlled transactivator generated by fusing the tet repressor with the activating domain of virion protein 16 of herpes simplex virus. tTA-IRES-Neo was generated by cloning PCR-amplified tTA cDNA from pUHD15-1 (Gossen, M. & Bujard, H., 1992, PNAS 89, 5547-5551) into the XbaI site of plasmid pMk10-59 (Kobayashi, M et al., 1996, Biotechniques 21, 398-402).
Proper citation: RRID:Addgene_16541 Copy
Species: Mus musculus
Genetic Insert: TOX2
Vector Backbone Description: Backbone Marker:Tannishtha Reya Lab; Backbone Size:6400; Vector Backbone:MSCV-IRES-eGFP; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_165428 Copy
Species: Homo sapiens
Genetic Insert: gRNA against human Total PGC-1a variants
Vector Backbone Description: Backbone Size:9580; Vector Backbone:pSico; Vector Types:Mammalian Expression, Lentiviral, CRISPR; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_165426 Copy
Species: Rattus norvegicus
Genetic Insert: iGluSnFR extracellular domain fused to TARP gamma-8 via NETO2 TM domain
Vector Backbone Description: Backbone Size:3950; Vector Backbone:pAAV; Vector Types:AAV; Bacterial Resistance:Ampicillin
References:
Comments: Chimera of iGluSnFR (Addgene plasmid # 41732), Neto2 and Gamma-8
pAAV backbone is based on Addgene plasmid # 61463
Please visit https://doi.org/10.1101/2021.01.21.427382 for bioRxiv preprint.
Proper citation: RRID:Addgene_165498 Copy
Species: Synthetic
Genetic Insert: CT81 gene circuit
Vector Backbone Description: Backbone Size:9311; Vector Backbone:pMMB206; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
References:
Comments: pMMB206 backbone is very low copy, <10 copies.
Proper citation: RRID:Addgene_165411 Copy
Species: Synthetic
Genetic Insert: Glyco-Venus-bPAC(S27A)-Myc
Vector Backbone Description: Backbone Size:3009; Vector Backbone:pGEM HE; Vector Types:Bacterial Expression, cDNA expression, Xenopus Oocyte; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_165490 Copy
Species: Homo sapiens
Genetic Insert: PPAR alpha
Vector Backbone Description: Backbone Marker:Amersham; Backbone Size:5000; Vector Backbone:pGEX-2TK; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_16550 Copy
Species: Homo sapiens
Genetic Insert: Human histone H2B
Vector Backbone Description: Backbone Marker:-; Backbone Size:8023; Vector Backbone:pRRL-cPPT-hPGKTMPrtTA-WPRE; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
References:
Comments: ABOUT BACKBONE:
The lentiviral backbone was originally designed and constructed by Barde I et al. (Barde I et al. Mol Ther. 2006. 13(2): 382-90. PMID: 16275162).
ABOUT TEST DIGESTION:
Since Xba I restriction site is lost during cloning, we recommend checking the plasmid using the enzymes: Xho I and Nhe I. You should get two fragments around 1600 bp (insert+vector) and 7500 bp (vector).
Proper citation: RRID:Addgene_165494 Copy
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