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Vector Backbone Description: Backbone Marker:Addgene plasmid 46881; Backbone Size:6733; Vector Backbone:pDL276; Vector Types:Bacterial Expression, Other, E. coli/Gram-positive Bacterial Shuttle Vector; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:1409970
Comments: This plasmid may contain unstable elements. Prolonged growth in E. coli selects for deletions, but not in E. faecalis. Test multiple clones upon to ensure the full plasmid is selected.
Proper citation: RRID:Addgene_46882 Copy
Vector Backbone Description: Backbone Size:8353; Vector Backbone:pMSP3535; Vector Types:Bacterial Expression, Nisin-inducible expression; Gram-positive Bacterial Shuttle Vector; Bacterial Resistance:Erythromycin
Defining Citation: PMID:10964628
Comments: This plasmid utilizes the NIsin Controlled Expression (NICE) system. NICE® is a registered trademark of NIZO food research BV, P.O.Box 20, 6710 BA Ede, The Netherlands.
pMSP3535 contains the Nisin-inducible PnisA promoter, and the pAMB1 replicon for expression in gram-positive bacteria.
For protein expression in E. faecalis, induce with 10-25 ng/mL nisin (Sigma, N-5764). See associated article for instructions on how to make nisin solution.
Note: Nucleotide sequence encoding the signal sequence and one-third of the mature NisA (nisin) peptide remains associated with the PnisA promoter. The depositing laboratory recommends adding of a Stop codon to the 5' end of inserted sequences to avoid unintentional gene fusions.
Addgene's quality control sequencing finds discrepancies with the depositor's provided sequencew which includes a mutation in RepE- F229L. This however is thought to not affect plasmid function.
Proper citation: RRID:Addgene_46886 Copy
Vector Backbone Description: Backbone Size:8278; Vector Backbone:pMSP3535VA; Vector Types:Bacterial Expression, Nisin-inducible expression; Gram-positive Bacterial Shuttle Vector; Bacterial Resistance:Kanamycin
Defining Citation: PMID:10964628
Comments: This plasmid utilizes the NIsin Controlled Expression (NICE) system. NICE® is a registered trademark of NIZO food research BV, P.O.Box 20, 6710 BA Ede, The Netherlands.
pMSP3535VA contains the Nisin-inducible PnisA promoter, and the pVA380-1 replicon for expression in gram-positive bacteria.
For protein expression in E. faecalis, induce with 10-25 ng/mL nisin (Sigma, N-5764). See associated article for instructions on how to make nisin solution.
Note: Nucleotide sequence encoding the signal sequence and one-third of the mature NisA (nisin) peptide remains associated with the PnisA promoter. The depositing laboratory recommends adding of a Stop codon to the 5' end of inserted sequences to avoid unintentional gene fusions.
Note: The theoretical full sequence uploaded here is NOT fully accurate - the NheI site at position 4414 has been lost, along with approximately 500bp of sequence in this region (compare the depositor's uploaded maps). The actual size of this plasmid is 8278 as listed here. The depositing lab has not resequenced the region in question, but the discrepancy does not affect plasmid function.
Note: This plasmid has been shown to accumulate deletions when replicated in E. coli (or gram-negative bacteria). Screening transformants by restriction digest is recommended.
Addgene's quality control sequencing finds discrepancies with the depositor's provided sequence, but they are not thought to affect plasmid function.
Proper citation: RRID:Addgene_46887 Copy
Genetic Insert: dCas9
Vector Backbone Description: Backbone Size:6900; Vector Backbone:pJED103; Vector Types:Yeast Expression, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:23849981
Comments: The Cas9 contains a N612K mutation that does not effect the function of the enzyme.
Proper citation: RRID:Addgene_46920 Copy
Vector Backbone Description: Backbone Marker:Homemade; Backbone Size:6990; Vector Backbone:pMA3211; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22637478
Proper citation: RRID:Addgene_46880 Copy
Genetic Insert: sgGFP-NT1
Vector Backbone Description: Backbone Marker:Addgene; Backbone Size:8200; Vector Backbone:pSICO derivative; Vector Types:Mammalian Expression, Lentiviral, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:23849981
Comments: gRNA target sequence GAATAGCTCAGAGGCCGAGG
Proper citation: RRID:Addgene_46914 Copy
Genetic Insert: sgGAL4-1
Vector Backbone Description: Backbone Marker:Addgene; Backbone Size:8200; Vector Backbone:pSICO derivative; Vector Types:Mammalian Expression, Lentiviral, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:23849981
Comments: gRNA target sequence GTTGGAGCACTGTCCTCCGAACGT
Proper citation: RRID:Addgene_46915 Copy
Vector Backbone Description: Backbone Marker:Homemade; Backbone Size:6642; Vector Backbone:pMA3211; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22637478
Proper citation: RRID:Addgene_46879 Copy
Species: Homo sapiens
Genetic Insert: dCas9-p65AD-BFP fusion
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:6300; Vector Backbone:MSCV-puro; Vector Types:Mammalian Expression, Retroviral, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:23849981
Proper citation: RRID:Addgene_46913 Copy
Species: Homo sapiens
Genetic Insert: sgCD71 -2
Vector Backbone Description: Backbone Marker:Addgene; Backbone Size:8200; Vector Backbone:pSICO derivative; Vector Types:Mammalian Expression, Lentiviral, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:23849981
Comments: gRNA target sequence GGACGCGCTAGTGTGAGTGC
Proper citation: RRID:Addgene_46918 Copy
Genetic Insert: sgGAL4-4
Vector Backbone Description: Backbone Marker:Addgene; Backbone Size:8200; Vector Backbone:pSICO derivative; Vector Types:Mammalian Expression, Lentiviral, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:23849981
Comments: gRNA target sequence GAACGACTAGTTAGGCGTGTA
Proper citation: RRID:Addgene_46916 Copy
Species: Mus musculus
Genetic Insert: becn1
Vector Backbone Description: Vector Backbone:pqcxih; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:23685627
Proper citation: RRID:Addgene_46994 Copy
Species: Rattus norvegicus
Genetic Insert: soluble guanylate cyclase 1alpha3
Vector Backbone Description: Backbone Marker:Homemade; Backbone Size:6642; Vector Backbone:pMA3211; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22637478
Comments: Please note that though there are some differences between Addgene's quality control sequence and the depositor's assembled sequence, this plasmid should function as reported.
Proper citation: RRID:Addgene_46874 Copy
Species: Mus musculus
Genetic Insert: becn1
Vector Backbone Description: Vector Backbone:pqcxih; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:23685627
Proper citation: RRID:Addgene_46995 Copy
Species: Drosophila melanogaster
Genetic Insert: Lis1
Vector Backbone Description: Vector Backbone:CS2; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:23097494
Proper citation: RRID:Addgene_47047 Copy
Species: Rattus norvegicus
Genetic Insert: soluble guanylate cyclase1 beta 3
Vector Backbone Description: Backbone Size:5684; Vector Backbone:pMA1662; Vector Types:Retroviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22637478
Proper citation: RRID:Addgene_46877 Copy
Species: Drosophila melanogaster
Genetic Insert: Asunder
Vector Backbone Description: Vector Backbone:CS2; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:23097494
Proper citation: RRID:Addgene_47041 Copy
Species: Drosophila melanogaster
Genetic Insert: Lis1
Vector Backbone Description: Vector Backbone:CS2; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:23097494
Proper citation: RRID:Addgene_47045 Copy
Species: Mus musculus
Genetic Insert: Asunder
Vector Backbone Description: Vector Backbone:CS2; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:23097494
Proper citation: RRID:Addgene_47044 Copy
Species: Homo sapiens
Genetic Insert: Tau E14 P301L
Vector Backbone Description: Backbone Marker:Clontech; Vector Backbone:pRK5; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21172610
Comments: WT htau construct (Addgene plasmid 46904) was used as a template to make mutations and contains human four-repeat tau lacking the N-terminal sequences (4R0N) and contains exons 1, 4 and 5, 7, and 9–13, intron 13, and exon 14.
Proper citation: RRID:Addgene_46909 Copy
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