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Species: Synthetic
Genetic Insert: 5'hSOX2se-Snrp-Tomato-polyA-Pgk-PURO-3'hSOX2se
Vector Backbone Description: Backbone Marker:invitrogen; Backbone Size:4583; Vector Backbone:topo; Vector Types:Mouse Targeting, Synthetic Biology; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26406378
Proper citation: RRID:Addgene_70155 Copy
Species: Synthetic
Genetic Insert: 5'hDAZL-snrp-GFP-polyA-pgK-PURO-3'hDAZL
Vector Backbone Description: Backbone Marker:invitrogen; Backbone Size:4258; Vector Backbone:topo; Vector Types:Mouse Targeting, Synthetic Biology; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26406378
Proper citation: RRID:Addgene_70144 Copy
Species: Synthetic
Genetic Insert: 5'hGapdh-Snrp-GFP-polyA-PGK-PURO-3'hGapdh
Vector Backbone Description: Backbone Marker:invitrogen; Backbone Size:3952; Vector Backbone:topo; Vector Types:Mouse Targeting, Synthetic Biology; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26406378
Proper citation: RRID:Addgene_70148 Copy
Species: Synthetic
Genetic Insert: alpha Sarcin (N-terminal portion)
Vector Backbone Description: Backbone Size:5428; Vector Backbone:pcDNA3.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24481070
Proper citation: RRID:Addgene_70230 Copy
Species: Synthetic
Genetic Insert: sfCherry(1-10)
Vector Backbone Description: Backbone Size:5500; Vector Backbone:pcDNA3.1(+); Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26988139
Proper citation: RRID:Addgene_70222 Copy
Species: Synthetic
Genetic Insert: amyEopt
Vector Backbone Description: Backbone Marker:Fagan and Fairweather 2009; Backbone Size:7216; Vector Backbone:pRPF185; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:27333161
Comments: Please note: the depositing lab has determined that a particular IS element present in E. coli strains such as DH5alpha and its derivatives has a high propensity to insert into this plasmid. The depositing lab recommends isolating the plasmid DNA and transforming into a strain similar to MC1061.
Publication describing the backbone vector pRPF185 as well as the sequencing primers:
Clostridium difficile has two parallel and essential Sec secretion systems. Fagan RP, Fairweather NF. J Biol Chem. 2011 Aug 5;286(31):27483-93. doi: 10.1074/jbc.M111.263889. Epub 2011 Jun 9.
PMID: 21659510
Proper citation: RRID:Addgene_70188 Copy
Species: Synthetic
Genetic Insert: eGFP
Vector Backbone Description: Backbone Size:5428; Vector Backbone:pcDNA3.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24481070
Proper citation: RRID:Addgene_70225 Copy
Species: Synthetic
Genetic Insert: alpha Sarcin (N-terminal portion)
Vector Backbone Description: Backbone Size:5428; Vector Backbone:pcDNA3.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24481070
Proper citation: RRID:Addgene_70229 Copy
Species: Synthetic
Genetic Insert: Cas9
Vector Backbone Description: Backbone Size:9080; Vector Backbone:pFUGW; Vector Types:Mammalian Expression, Lentiviral, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26375006
Comments: Subcloned Venus in place of Blast in lentiCas9-Blast.
Improved vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nat Methods. 2014 Aug;11(8):783-4. doi: 10.1038/nmeth.3047. 10.1038/nmeth.3047 PubMed 25075903
Canver MC, Smith EC, Sher F, Pinello L, Sanjana NE, Shalem O, Chen DD, Schupp PG, Vinjamur DS, Garcia SP, Luc S, Kurita R, Nakamura Y, Fujiwara Y, Maeda T, Yuan G, Zhang F, Orkin SH, Bauer DE. BCL11A enhancer dissection by Cas9-mediated in situ saturating mutagenesis. Nature. 2015 September 16; in press; doi:10.1038/nature15521.
Proper citation: RRID:Addgene_70267 Copy
Species: Synthetic
Genetic Insert: S. pyogenes sgRNA cassette
Vector Backbone Description: Backbone Size:9482; Vector Backbone:Custom; Vector Types:Mammalian Expression, Lentiviral, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26375006
Comments: Cloned in Crimson Reporter in place of PURO reporter into lentiGuide-Puro.
Improved vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nat Methods. 2014 Aug;11(8):783-4. doi: 10.1038/nmeth.3047. 10.1038/nmeth.3047 PubMed 25075903
Canver MC, Smith EC, Sher F, Pinello L, Sanjana NE, Shalem O, Chen DD, Schupp PG, Vinjamur DS, Garcia SP, Luc S, Kurita R, Nakamura Y, Fujiwara Y, Maeda T, Yuan G, Zhang F, Orkin SH, Bauer DE. BCL11A enhancer dissection by Cas9-mediated in situ saturating mutagenesis. Nature. 2015 September 16; in press; doi:10.1038/nature15521.
Proper citation: RRID:Addgene_70683 Copy
Species: Synthetic
Genetic Insert: Channelrhodopsin-2
Vector Backbone Description: Backbone Size:5601; Vector Backbone:pAAV; Vector Types:Mammalian Expression, AAV; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26443033
Proper citation: RRID:Addgene_70762 Copy
Species: Synthetic
Genetic Insert: bar
Vector Backbone Description: Backbone Size:7132; Vector Backbone:pPZP201; Vector Types:Plant Expression; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:15034747
Comments: This vector is suitable for plant transformation and has been used as standard service empty vector control.
Proper citation: RRID:Addgene_73933 Copy
Species: Synthetic
Genetic Insert: bar
Vector Backbone Description: Backbone Size:7132; Vector Backbone:pPZP201; Vector Types:Plant Expression; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:15034747
Comments: This vector is suitable hpRNA-mediated gene silencing in tissue (seed) specific manner.
Please see the Plant Transformation Core Facility website for more information on using this plasmid: http://plantsci.missouri.edu/muptcf/pzy101.html
The vector has following features:
1. It has two replicate origins that allow to replicate in both E. coli and Agrobacterium tumefaciens cells (at higher copy numbers).
2. It also has bom site that allows you to use tri-parental mating to mobilize the vector to Agrobacterium cells.
3. The aadA locus confers Spectinomycin (100 mg/L) or streptomycin (100 mg/L) resistance and thus enable to use either or both of these two antibiotics for the vector selection.
4. The multiple cloning site (MCS) adjacent to the double CaMV35S promoter will allow you to insert your cassette. This CaMV35S promoter is used to drive the bar ORF and, therefore, is not used in front to drive your gene of interest. You need to assemble your own expression cassette including promoter, gene coding sequence, and terminator, and then insert to anywhere between Hind III and EcoR I within the MCS.
Special notes:
1. Insert your gene expression cassette in such a way that your promoter is adjacent to the promoter driving the bar gene selectable marker. This is to prevent transcription read-through from your gene expression cassette.
2. In order to use the EcoR I site within the MCS, a minimal ratio of this enzyme versus DNA is recommended to avoid star activity. When HindIII and EcoRI are in combined use, even less of each enzyme is required, and reaction should be conducted in a large volume to avoid excessive enzymatic activity.
Proper citation: RRID:Addgene_73932 Copy
Species: Synthetic
Genetic Insert: Golden Gate destination vector - Level 2, 2 TU's with secondary module and colE1 origin
Vector Backbone Description: Backbone Marker:Terrence Lai (this study); Backbone Size:1878; Vector Backbone:colE1 origin and chloramphenicol resistance; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:27096716
Proper citation: RRID:Addgene_74091 Copy
Species: Synthetic
Genetic Insert: TL9 RBS
Vector Backbone Description: Backbone Marker:iGEM; Backbone Size:2044; Vector Backbone:pSB1C3; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:27096716
Comments: This plasmid contains the TL9 RBS upstream of GFP.
Proper citation: RRID:Addgene_74098 Copy
Species: Synthetic
Genetic Insert: Golden Gate destination vector - Level 2, 3 TU's with secondary module and p15A origin
Vector Backbone Description: Backbone Marker:Terrence Lai (this study); Backbone Size:2024; Vector Backbone:p15A origin and chloramphenicol resistance; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:27096716
Proper citation: RRID:Addgene_74089 Copy
Species: Synthetic
Genetic Insert: sgRNA pyk (NT)
Vector Backbone Description: Vector Backbone:pAL374; Vector Types:Bacterial Expression, CRISPR; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:26829286
Proper citation: RRID:Addgene_74072 Copy
Species: Synthetic
Genetic Insert: sgRNA pyk (T)
Vector Backbone Description: Vector Backbone:pAL374; Vector Types:Bacterial Expression, CRISPR, Synthetic Biology; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:26829286
Proper citation: RRID:Addgene_74070 Copy
Species: Synthetic
Genetic Insert: sgRNA rfp (T)
Vector Backbone Description: Vector Backbone:pAL374; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:26829286
Proper citation: RRID:Addgene_74075 Copy
Species: Synthetic
Genetic Insert: MBIC5-mCer3
Vector Backbone Description: Backbone Size:4095; Vector Backbone:pCS2+; Vector Types:Mammalian Expression, mRNA generation for microinjection.; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26808669
Proper citation: RRID:Addgene_74117 Copy
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