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Species: Streptomyces noursei
Genetic Insert: natMX4
Vector Backbone Description: Vector Backbone:pFA6; Vector Types:Yeast replicating; Bacterial Resistance:Ampicillin
Defining Citation: PMID:10514571
Proper citation: RRID:Addgene_35121 Copy
Vector Backbone Description: Backbone Marker:Ji-Wu Wang and Brian McCabe; Backbone Size:10033; Vector Backbone:pBID-G; Vector Types:Drosophila Transgene Expression; Bacterial Resistance:Chloramphenicol and Ampicillin
Defining Citation: PMID:22848718
Comments: Generation of ϕC31 transgenic Drosophila. Gateway cloning cassette is flanked by gypsy insulator sequences to allow uniform transgene expression between insertion sites. Transgene selection using white gene. loxP site facilitates elimination of transgene markers via Cre recombinase-mediated excision with ZH-attP landing sites. Ampicillin resistant. Chloramphenicol resistant for amplification.
Please see the associated publication for more information: Wang J-W, Beck ES, McCabe BD (2012) A Modular Toolset for Recombination Transgenesis and Neurogenetic Analysis of Drosophila. PLoS ONE 7(7): e42102 http://www.plosone.org/article/info:doi%2F10.1371%2Fjournal.pone.0042102
Proper citation: RRID:Addgene_35195 Copy
Species: Synthetic
Genetic Insert: PCP-2x-yeGFP
Vector Backbone Description: Vector Backbone:pURA; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21512033
Comments: This plasmid expresses a chimeric RNA-binding protein fused to GFP used to detect mRNAs that contain multiple copies of an RNA sequence (see Addgene plasmid #'s 35191, 35192 & 35196). See associated publication for detailed methods and use of this plasmid.
Please note that Addgene's sequencing results identified discrepancies when compared to the assembled sequence provided by the depositing laboratory. The first GFP construct in this plasmid (bp#441-1154) is yeast codon optimized and contains a E124K mutation and amino acid 206 is A (not K, as suggested by the depositor's sequence). The second GFP construct in this plasmid (bp# 1212-1928) is NOT yeast codon optimized.
This construct has successfully been used by the depositing laboratory as detailed in the associated publication and the sequencing discrepancies are not a concern for the function of the plasmid.
Proper citation: RRID:Addgene_35194 Copy
Species: Synthetic
Genetic Insert: GLT1 POL1 24xPP7SL
Vector Backbone Description: Vector Backbone:pHIS; Vector Types:; Bacterial Resistance:Ampicillin and Kanamycin
Defining Citation: PMID:21512033
Comments: Note that Addgene NGS results found 22xPP7 stem loop repeats rather than expected 24. It is not known how this impacts plasmid function.
This plasmid was designed to construct mRNAs that contain multiple copies of an RNA sequence (the 24xPP7 stem-loops) that can be recognized by a chimeric RNA-binding protein fused to GFP, encoded in plasmid pDZ276 (Addgene plasmid #35194). See associated publication for detailed methods and use of this plasmid.
Please note that the LacZa-ccdB fusion in this plasmid is not functional and was originally present in the vector backbone to aid in screening for insert.
Proper citation: RRID:Addgene_35196 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: fcy1PR
Vector Backbone Description: Vector Backbone:pFA6-kanMX3; Vector Types:Yeast replicating; Bacterial Resistance:Ampicillin
Defining Citation: PMID:16088873
Proper citation: RRID:Addgene_35104 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: fcy1MX4
Vector Backbone Description: Vector Backbone:pFA6-kanMX4; Vector Types:Yeast replicating; Bacterial Resistance:Ampicillin
Defining Citation: PMID:16088873
Proper citation: RRID:Addgene_35102 Copy
Species: Homo sapiens
Genetic Insert: PSF
Vector Backbone Description: Backbone Size:3800; Vector Backbone:pCMVmyc; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:16024807
Proper citation: RRID:Addgene_35183 Copy
Vector Backbone Description: Backbone Marker:Ji-Wu Wang and Brian McCabe; Backbone Size:8286; Vector Backbone:pBID; Vector Types:Drosophila Transgene Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22848718
Comments: Generation of ϕC31transgenic Drosophila. Multiple cloning site is flanked by gypsy insulator sequences to allow uniform transgene expression between insertion sites. MCS shares similarity to pUAST. Transgene selection using white gene. loxP site facilitates elimination of transgene markers via Cre recombinase-mediated excision with ZH-attP landing sites. Ampicillin resistant.
Please see the associated publication for more information: Wang J-W, Beck ES, McCabe BD (2012) A Modular Toolset for Recombination Transgenesis and Neurogenetic Analysis of Drosophila. PLoS ONE 7(7): e42102 http://www.plosone.org/article/info:doi%2F10.1371%2Fjournal.pone.0042102
Proper citation: RRID:Addgene_35190 Copy
Species: Homo sapiens
Genetic Insert: RPBI
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5400; Vector Backbone:pcDNA3.1(-); Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15037767
Comments: α-amanitin-resistant form of the largest subunit of pol II
Proper citation: RRID:Addgene_35176 Copy
Species: Mus musculus
Genetic Insert: MmnSR100
Vector Backbone Description: Backbone Size:7032; Vector Backbone:pLKO.1-puro; Vector Types:Mammalian Expression, Lentiviral, RNAi; Bacterial Resistance:Ampicillin
Defining Citation: PMID:19737518
Proper citation: RRID:Addgene_35174 Copy
Species: Synthetic
Genetic Insert: RFP
Vector Backbone Description: Backbone Size:4904; Vector Backbone:pBb; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:21933410
Comments: JBEI Part ID: JPUB_000050; Origin: SC101
Proper citation: RRID:Addgene_35284 Copy
Species: Synthetic
Genetic Insert: RFP
Vector Backbone Description: Backbone Size:3476; Vector Backbone:pBb; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:21933410
Comments: JBEI Part ID: JPUB_000056; Origin: p15A
Proper citation: RRID:Addgene_35290 Copy
Species: Synthetic
Genetic Insert: RFP
Vector Backbone Description: Backbone Size:3596; Vector Backbone:pBb; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21933410
Comments: JBEI Part ID: JPUB_000081; Origin: colE1
Proper citation: RRID:Addgene_35315 Copy
Species: Synthetic
Genetic Insert: RFP
Vector Backbone Description: Backbone Size:3281; Vector Backbone:pBb; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21933410
Comments: JBEI Part ID: JPUB_000036; Origin: colE1
Proper citation: RRID:Addgene_35270 Copy
Species: Synthetic
Genetic Insert: RFP
Vector Backbone Description: Backbone Size:3340; Vector Backbone:pBb; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21933410
Comments: JBEI Part ID: JPUB_000034; Origin: colE1
Proper citation: RRID:Addgene_35268 Copy
Species: Synthetic
Genetic Insert: RFP
Vector Backbone Description: Backbone Size:5360; Vector Backbone:pBb; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21933410
Comments: JBEI Part ID: JPUB_000067; Origin: SC101
Proper citation: RRID:Addgene_35301 Copy
Species: Danio rerio
Genetic Insert: Zinc finger array targeting zgc:111821
Vector Backbone Description: Backbone Size:5493; Vector Backbone:MG414; Vector Types:Zebrafish Targeting; Bacterial Resistance:Kanamycin
Defining Citation: PMID:18657511
Comments: This plasmid encodes a zinc finger array targeting half of a target sequence in the zebrafish gene zgc:111821. Please note that this plasmid does NOT contain the zgc:111821 sequence.
Users must order the complementary plasmid zgc:111821_R (OZ598) [Addgene plasmid 35238] in order to create a zinc finger nuclease (ZFN) pair to introduce targeted mutations into this specific zebrafish gene.
Users will also need to clone the zinc finger (ZF) insert of this plasmid into a zinc finger nuclease (ZFN) vector to express a FOKI fusion product. Examples of possible ZFN expression vectors that can be used are: pST1374 (Addgene plasmid 13426), pMLM290/292 (Addgene plasmids 21872 & 21873), pMLM800/802 (Addgene plasmids 27202 & 27203).
The difference between pMLM800/802 and pMLM290/292 is the length of the “spacer” sequence in the full ZFN target site. If the spacer is 7 bp, scientists should use pMLM800/802. If the spacer is 5 or 6 bps, scientists should use pMLM290/292.
pMLM290/292 is identical to pST1374 except that it harbors two mutations in the FokI nuclease domain (Q486E, I499L; aka the “-“ mutation see Miller et al., Nat. Biotech 2007, PMID 17603475) which confers heterodimeric behavior on these domains
This zinc finger array was tested for binding activity to the sequence 5'-GCTGCTGCC-3' in a bacterial two hybrid assay, and resulted in 3.61 fold activation. However, this array has not yet been tested for activity as a zinc finger nuclease (i.e. for its ability to induce mutations at the intended locus).
Scientists using this zinc finger array in a publication should notify [email protected] and acknowledge NIH grant number R01 GM088040 in the publication.
Other Articles: "Oligomerized pool engineering (OPEN): an 'open-source' protocol for making customized zinc-finger arrays." Maeder ML et al. (Nat Protoc. 2009 Sept 17. 4(10):1471-1501. Pubmed ID: 19798082)
"Targeted mutagenesis in zebrafish using customized zinc-finger nucleases." Foley JE et al. (Nat Protoc. 2009 Dec 3. 4(12):1855-1867. Pubmed ID: 20010934)
Proper citation: RRID:Addgene_35237 Copy
Species: Synthetic
Genetic Insert: RFP
Vector Backbone Description: Backbone Size:2777; Vector Backbone:pBb; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21933410
Comments: JBEI Part ID: JPUB_000090; Origin: colE1
Proper citation: RRID:Addgene_35324 Copy
Species: Synthetic
Genetic Insert: RFP
Vector Backbone Description: Backbone Size:2764; Vector Backbone:pBb; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21933410
Comments: JBEI Part ID: JPUB_000093; Origin: p15A
Proper citation: RRID:Addgene_35327 Copy
Species: Synthetic
Genetic Insert: RFP
Vector Backbone Description: Backbone Size:2600; Vector Backbone:pBb; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:21933410
Comments: JBEI Part ID: JPUB_000092; Origin: p15A
Proper citation: RRID:Addgene_35326 Copy
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