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http://www.addgene.org/105807

Vector Backbone Description: Backbone Marker:Sammarco & Grabczyk (PMID: 16212928); Vector Backbone:BI-16 (constructed based on pcDNA5/FRT/TO from Invitrogen); Vector Types:Mammalian Expression, RNAi, Flp-In competent; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29590189
Comments: This vector is a member of the pKK-RNAi series which belongs to the pKK vector family. It enables concomitant expression of two genes: 1) a cassette encoding miRNAs that target the gene of interest, and 2) an allele of the gene of interest with the protein coding sequence harbouring silent mutations that make the mRNA insensitive to the miRNAs. As a result, the endogenous alleles of the gene of interest are silenced, whereas the exogenous copy is expressed. Furthermore, the miRNAs are cotranscriptionally expressed with a fluorescent protein reporter so that analysis can be narrowed down to cells that express miRNA, provided that applied assay can distinguish cells expressing reporter. The reporter of miRNA expression has 3 repeats of the nuclear localisation signal of SV40 large T antigen, thus, it localizes to nucleus. This vector is a derivate of the BI16 plasmid (PMID: 16212928) and is suitable for stable cell line generation using the Flp-In system. Transcription of cloned DNA is driven by a tetracycline responsive bidirectional CMV promoter (see Szczesny RJ et al. for detailed description and protocols; bioRxiv, doi: https://doi.org/10.1101/160101).

Proper citation: RRID:Addgene_105807 Copy   

•   Source: Addgene
  

http://www.addgene.org/105804

Vector Backbone Description: Backbone Marker:Sammarco & Grabczyk (PMID: 16212928); Vector Backbone:BI-16 (constructed based on pcDNA5/FRT/TO from Invitrogen); Vector Types:Mammalian Expression, Flp-In competent; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29590189
Comments: Addgene has found the VN173 mutation (I152L) is not present in this plasmid. This vector is a member of the pKK vector family. It enables concomitant expression of two genes, which transcription is driven by a tetracycline responsive bidirectional CMV promoter. This vector is a derivate of the BI16 plasmid (PMID: 16212928) and is suitable for stable cell line generation using the Flp-In system (see Szczesny RJ et al. for detailed description and protocols; bioRxiv, doi: https://doi.org/10.1101/160101). pKK-BiFC-Venus vector encodes fragments of Venus (VN173, VC155) that become fluorescent when they reassemble into full fluorescent protein. Thus, it is useful for protein-protein interactions studies involving bimolecular fluorescence complementation approach (BiFC) (PMID: 16454041, 18846096). Here, an improved version of VN173 fragment is used (I152L) to inhibit self-assembly (PMID:21091444). Venus fragments coding sequences were cloned from Addgene ID: 22010 and 22011 and VN173 was subjected to site-directed mutagenesis to introduce mutation I152L.

Proper citation: RRID:Addgene_105804 Copy   

•   Source: Addgene
  

http://www.addgene.org/105802

Vector Backbone Description: Backbone Marker:Sammarco & Grabczyk (PMID: 16212928); Vector Backbone:BI-16 (constructed based on pcDNA5/FRT/TO from Invitrogen); Vector Types:Mammalian Expression, Flp-In competent; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29590189
Comments: This vector is a member of the pKK-BI16 series which belongs to the pKK vector family. It enables concomitant expression of two genes, which transcription is driven by a tetracycline responsive bidirectional CMV promoter. This vector is a derivate of the BI16 plasmid (PMID: 16212928) and is suitable for stable cell line generation using the Flp-In system. ORF2 is cloned with the universal SLIC protocol (just three universal PCR primers are required to clone a given coding sequence into all vectors from pKK family by a ligation-independent DNA cloning method), ORF1 can be cloned using SLIC with specifically designed primers (applicable enzymes: Bsp120I, ApaI, MluI, NotI, BspTI, Eco47III, MunI) or conventional restriction-ligase based cloning (see Szczesny RJ et al. for detailed description and protocols; bioRxiv, doi: https://doi.org/10.1101/160101). mClover3 is a brighter derivative of mEGFP. mRuby3 is brighter than mCherry and has slightly shifted excitation and emission spectra. The mClover3-mRuby3 pair is superior to EGFP-mCherry in FRET analysis (PMID: 26879144). Useful for experiments requiring co-expression of two proteins (e.g. FRET measurements).

Proper citation: RRID:Addgene_105802 Copy   

•   Source: Addgene
  

http://www.addgene.org/105803

Vector Backbone Description: Backbone Marker:Sammarco & Grabczyk (PMID: 16212928); Vector Backbone:BI-16 (constructed based on pcDNA5/FRT/TO from Invitrogen); Vector Types:Mammalian Expression, Flp-In competent; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29590189
Comments: This vector is a member of the pKK-BI16 series which belongs to the pKK vector family. It enables concomitant expression of two genes, which transcription is driven by a tetracycline responsive bidirectional CMV promoter. This vector is a derivate of the BI16 plasmid (PMID: 16212928) and is suitable for stable cell line generation using the Flp-In system. ORF2 is cloned with the universal SLIC protocol (just three universal PCR primers are required to clone a given coding sequence into all vectors from pKK family by a ligation-independent DNA cloning method), ORF1 can be cloned using SLIC with specifically designed primers (applicable enzymes: Bsp120I, ApaI, MluI, NotI, BspTI, Eco47III, MunI, Psp5II) or conventional restriction-ligase based cloning (see Szczesny RJ et al. for detailed description and protocols; bioRxiv, doi: https://doi.org/10.1101/160101). mClover3 is a brighter derivative of mEGFP (PMID: 26879144). FLAG is a short tag, less likely to interfere with protein function (PMID: 11694294).

Proper citation: RRID:Addgene_105803 Copy   

•   Source: Addgene
  

http://www.addgene.org/105800

Vector Backbone Description: Backbone Marker:Sammarco & Grabczyk (PMID: 16212928); Vector Backbone:BI-16 (constructed based on pcDNA5/FRT/TO from Invitrogen); Vector Types:Mammalian Expression, Flp-In competent; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29590189
Comments: This vector is a member of the pKK-BI16 series which belongs to the pKK vector family. It enables concomitant expression of two genes, which transcription is driven by a tetracycline responsive bidirectional CMV promoter. This vector is a derivate of the BI16 plasmid (PMID: 16212928) and is suitable for stable cell line generation using the Flp-In system. One coding sequence is cloned with the universal SLIC protocol (just three universal PCR primers are required to clone a given coding sequence into all vectors from pKK family by a ligation-independent DNA cloning method), the other one can be cloned using SLIC with specifically designed primers (applicable enzymes: Bsp120I, ApaI, MluI, NotI, BspTI, Eco47III, MunI, Psp5II) or conventional restriction-ligase based cloning (see Szczesny RJ et al. for detailed description and protocols; bioRxiv, doi: https://doi.org/10.1101/160101).

Proper citation: RRID:Addgene_105800 Copy   

•   Source: Addgene
  

http://www.addgene.org/105801

Vector Backbone Description: Backbone Marker:Sammarco & Grabczyk (PMID: 16212928); Vector Backbone:BI-16 (constructed based on pcDNA5/FRT/TO from Invitrogen); Vector Types:Mammalian Expression, Flp-In competent; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29590189
Comments: This vector is a member of the pKK-BI16 series which belongs to the pKK vector family. It enables concomitant expression of two genes, which transcription is driven by a tetracycline responsive bidirectional CMV promoter. This vector is a derivate of the BI16 plasmid (PMID: 16212928) and is suitable for stable cell line generation using the Flp-In system. ORF2 is cloned with the universal SLIC protocol (just three universal PCR primers are required to clone a given coding sequence into all vectors from pKK family by a ligation-independent DNA cloning method), ORF1 can be cloned using SLIC with specifically designed primers (applicable enzymes: MluI, BspTI, Eco47III, MunI, Psp5II) or conventional restriction-ligase based cloning (see Szczesny RJ et al. for detailed description and protocols; bioRxiv, doi: https://doi.org/10.1101/160101). mClover3 is a brighter derivative of mEGFP (PMID: 26879144). FLAG is a short tag, less likely to interfere with protein function (PMID: 11694294).

Proper citation: RRID:Addgene_105801 Copy   

•   Source: Addgene
  

http://www.addgene.org/105922

Genetic Insert: mKate2
Vector Backbone Description: Vector Backbone:pAAV; Vector Types:AAV; Bacterial Resistance:Ampicillin
Comments: Entire transgene cassette can be removed with NotI/XbaI digest; CBh promoter can be removed by XbaI/AgeI digest; mKate2 can be removed with SaI/EcoRI digest; IRES can be removed by EcoRI/FseI digest

Proper citation: RRID:Addgene_105922 Copy   

•   Source: Addgene
  

http://www.addgene.org/105851

Species: Mus musculus
Genetic Insert: IgG1 heavy chain, IgG3 heavy chain
Vector Backbone Description: Backbone Marker:Invivogen; Backbone Size:4483; Vector Backbone:pFUSEss-CHIg-mG1 (modified); Vector Types:Mammalian Expression; Bacterial Resistance:Bleocin (Zeocin)
Defining Citation: PMID:29875771

Proper citation: RRID:Addgene_105851 Copy   

•   Source: Addgene
  

http://www.addgene.org/105852

Species: Mus musculus
Genetic Insert: IgG1 heavy chain
Vector Backbone Description: Backbone Marker:Invivogen; Backbone Size:4474; Vector Backbone:pFUSEss-CHIg-mG1 (modified); Vector Types:Mammalian Expression; Bacterial Resistance:Bleocin (Zeocin)
Defining Citation: PMID:29875771

Proper citation: RRID:Addgene_105852 Copy   

•   Source: Addgene
  

http://www.addgene.org/105850

Species: Mus musculus
Genetic Insert: IgG1 heavy chain, IgG3 heavy chain
Vector Backbone Description: Backbone Marker:Invivogen; Backbone Size:4483; Vector Backbone:pFUSEss-CHIg-mG1 (modified); Vector Types:Mammalian Expression; Bacterial Resistance:Bleocin (Zeocin)
Defining Citation: PMID:29875771

Proper citation: RRID:Addgene_105850 Copy   

•   Source: Addgene
  

http://www.addgene.org/105858

Species: Mus musculus
Genetic Insert: IgG3 heavy chain
Vector Backbone Description: Backbone Marker:Invivogen; Backbone Size:4483; Vector Backbone:pFUSEss-CHIg-mG3 (modified); Vector Types:Mammalian Expression; Bacterial Resistance:Bleocin (Zeocin)
Defining Citation: PMID:29875771

Proper citation: RRID:Addgene_105858 Copy   

•   Source: Addgene
  

http://www.addgene.org/105856

Species: Mus musculus
Genetic Insert: IgG1, IgG3
Vector Backbone Description: Backbone Marker:Invivogen; Backbone Size:4474; Vector Backbone:pFUSEss-CHIg-mG3 (modified); Vector Types:Mammalian Expression; Bacterial Resistance:Bleocin (Zeocin)
Defining Citation: PMID:29875771

Proper citation: RRID:Addgene_105856 Copy   

•   Source: Addgene
  

http://www.addgene.org/105854

Species: Mus musculus
Genetic Insert: IgG1 heavy chain, IgG3 heavy chain
Vector Backbone Description: Backbone Marker:Invivogen; Backbone Size:4483; Vector Backbone:pFUSEss-CHIg-mG1 (modified); Vector Types:Mammalian Expression; Bacterial Resistance:Bleocin (Zeocin)
Defining Citation: PMID:29875771

Proper citation: RRID:Addgene_105854 Copy   

•   Source: Addgene
  

http://www.addgene.org/105862

Species: Mus musculus
Genetic Insert: IgG3 heavy chain
Vector Backbone Description: Backbone Marker:Invivogen; Backbone Size:4483; Vector Backbone:pFUSEss-CHIg-mG3; Vector Types:Mammalian Expression; Bacterial Resistance:Bleocin (Zeocin)
Defining Citation: PMID:29875771

Proper citation: RRID:Addgene_105862 Copy   

•   Source: Addgene
  

http://www.addgene.org/105860

Species: Mus musculus
Genetic Insert: IgG3 F(ab')2
Vector Backbone Description: Backbone Marker:Invivogen; Backbone Size:4549; Vector Backbone:pFUSEss-CHIg-mG3 (modified); Vector Types:Mammalian Expression; Bacterial Resistance:Bleocin (Zeocin)
Defining Citation: PMID:29875771

Proper citation: RRID:Addgene_105860 Copy   

•   Source: Addgene
  

http://www.addgene.org/105868

Species: Homo sapiens
Genetic Insert: Hspa1a (synonym Hsp68) MiniPromoter
Vector Backbone Description: Vector Backbone:unknown; Vector Types:; Bacterial Resistance:Ampicillin
Defining Citation: PMID:30765420
Comments: Please see the annotated GenBank file and the additional image file for the specific location of features in this plasmid, including the MiniPromoter.

Proper citation: RRID:Addgene_105868 Copy   

•   Source: Addgene
  

http://www.addgene.org/105866

Species: Arabidopsis thaliana
Genetic Insert: U6-26p::sgRNA scaffold
Vector Backbone Description: Backbone Marker:Promega; Backbone Size:3017; Vector Backbone:pGEM T-easy; Vector Types:Plant Expression, CRISPR, Synthetic Biology, Subcloning; Bacterial Resistance:Ampicillin
Defining Citation: PMID:34541122
Comments: The new version of pFH6 (pFH6_new) provides > 10 times higher mutation efficiencies than the original version. In combination with pUB-Cas9, homozygous knockout plants can be created in Arabidopsis in the T2 generation at efficiencies of 50-100%. For cloning procedure, please use the attached protocol which is derived from Hahn et al., 2017 ( http://www.bio-protocol.org/e2384 ).

Proper citation: RRID:Addgene_105866 Copy   

•   Source: Addgene
  

http://www.addgene.org/105865

Species: Synthetic
Genetic Insert: K-GECO1
Vector Backbone Description: Vector Backbone:pcDNA3.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29338710
Comments: bioRxiv--A genetically encoded Ca2+ indicator based on circularly permutated sea anemone red fluorescent protein (https://doi.org/10.1101/213082)

Proper citation: RRID:Addgene_105865 Copy   

•   Source: Addgene
  

http://www.addgene.org/105839

Species: E. coli
Genetic Insert: hydroperoxidase II catalase
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:4100; Vector Backbone:pBad; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: Please visit https://openlibrary-repo.ecampusontario.ca/jspui/handle/123456789/838 for Chemical Biology & Biochemistry Laboratory Using Genetic Code Expansion Manual.

Proper citation: RRID:Addgene_105839 Copy   

•   Source: Addgene
  

http://www.addgene.org/105796

Vector Backbone Description: Backbone Marker:Invitrogen; Vector Backbone:pcDNA5/FRT/TO; Vector Types:Mammalian Expression, Flp-In competent; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29590189
Comments: This vector is a member of the pKK vector family. Its cloning site was optimized for the SLIC approach, however, a traditional restriction and ligation based approach is also possible (see Szczesny RJ et al. for detailed protocols; bioRxiv, doi: https://doi.org/10.1101/160101). This vector is a derivate of the pcDNA5/FRT/TO plasmid (Invitrogen) and is suitable for stable cell line generation using the Flp-In system. Transcription of the gene of interest is driven by a tetracycline responsive CMV promoter. A given coding sequence can be cloned into all pKK vectors using just three universal PCR primers and a ligation-independent DNA cloning method. mCherry descends from the first truly monomeric red fluorescent protein mRFP1, which was obtained by mutating DsRed form Discosoma sp. Note that EGFP and mCherry come from different organisms and their sequences differ greatly. However, GFP-type termini were introduced into mCherry to increase tolerance of N- and C-terminal fusions in mammalian cells (PMID: 15558047). Thus, terminal sequences of EGFP and mCherry are the same.

Proper citation: RRID:Addgene_105796 Copy   

•   Source: Addgene