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Species: Yarrowia lipolytica
Genetic Insert: Contains gRNA targeting TF27 (YALI1_E22758g) and homologous arm matching TF27
Vector Backbone Description: Vector Backbone:Our own backbone; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:40460129
Comments: This plasmid contains gRNA targeting TF27 (YALI1_E22758g) and homologous arm matching TF27, which can be used for library construction by inserting promoters using a golden gate or directly knocking TF27 out.
Proper citation: RRID:Addgene_219892 Copy
Species: Yarrowia lipolytica
Genetic Insert: Promoter FBA
Vector Backbone Description: Vector Backbone:TOPO Vector; Vector Types:Yeast Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:40460129
Proper citation: RRID:Addgene_219925 Copy
Species: Yarrowia lipolytica
Genetic Insert: Contains gRNA targeting TF19 (YALI1_A20855g) and homologous arm matching TF19
Vector Backbone Description: Vector Backbone:Our own backbone; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:40460129
Comments: This plasmid contains gRNA targeting TF19 (YALI1_A20855g) and homologous arm matching TF19, which can be used for library construction by inserting promoters using a golden gate or directly knocking TF19 out.
Proper citation: RRID:Addgene_219884 Copy
Species: Yarrowia lipolytica
Genetic Insert: Contains gRNA targeting TF18 (YALI1_D15792g) and homologous arm matching TF18
Vector Backbone Description: Vector Backbone:Our own backbone; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:40460129
Comments: This plasmid contains gRNA targeting TF18 (YALI1_D15792g) and homologous arm matching TF18, which can be used for library construction by inserting promoters using a golden gate or directly knocking TF18 out.
Proper citation: RRID:Addgene_219883 Copy
Species: Homo sapiens
Genetic Insert: Sp-nCas9(D10A)-SpRY_P2A-mCherry
Vector Backbone Description: Vector Backbone:pCMV; Vector Types:Mammalian Expression, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:38773305
Comments: QTC290; could also be used for dual-nicking or other editing modalities
Proper citation: RRID:Addgene_219935 Copy
Species: Mus musculus
Genetic Insert: IgK
Vector Backbone Description: Vector Backbone:pAPtag5; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:38574730
Proper citation: RRID:Addgene_219813 Copy
Species: Homo sapiens
Genetic Insert: Sp-nCas9(D10A)-NG_P2A-mCherry
Vector Backbone Description: Vector Backbone:pCMV; Vector Types:Mammalian Expression, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:38773305
Comments: QTC24; could also be used for dual-nicking or other editing modalities
Proper citation: RRID:Addgene_219934 Copy
Species: Yarrowia lipolytica
Genetic Insert: Contains gRNA targeting TF29 (YALI1_C09133g) and homologous arm matching TF29
Vector Backbone Description: Vector Backbone:Our own backbone; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:40460129
Comments: This plasmid contains gRNA targeting TF29 (YALI1_C09133g) and homologous arm matching TF29, which can be used for library construction by inserting promoters using a golden gate or directly knocking TF29 out.
Proper citation: RRID:Addgene_219894 Copy
Species: Mus musculus
Genetic Insert: IgK
Vector Backbone Description: Vector Backbone:pZac2.1; Vector Types:Mammalian Expression, AAV; Bacterial Resistance:Ampicillin
Defining Citation: PMID:38574730
Proper citation: RRID:Addgene_219811 Copy
Species: Yarrowia lipolytica
Genetic Insert: Contains gRNA targeting TF34 (YALI1_E37455g) and homologous arm matching TF34
Vector Backbone Description: Vector Backbone:Our own backbone; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:40460129
Comments: This plasmid contains gRNA targeting TF34 (YALI1_E37455g) and homologous arm matching TF34, which can be used for library construction by inserting promoters using a golden gate or directly knocking TF34 out.
Proper citation: RRID:Addgene_219899 Copy
Species: Mus musculus
Genetic Insert: IgK
Vector Backbone Description: Vector Backbone:pZac2.1; Vector Types:Mammalian Expression, AAV; Bacterial Resistance:Ampicillin
Defining Citation: PMID:38574730
Proper citation: RRID:Addgene_219810 Copy
Species: Yarrowia lipolytica
Genetic Insert: Contains gRNA targeting TF33 (YALI1_F14650g) and homologous arm matching TF33
Vector Backbone Description: Vector Backbone:Our own backbone; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:40460129
Comments: This plasmid contains gRNA targeting TF33 (YALI1_F14650g) and homologous arm matching TF33, which can be used for library construction by inserting promoters using a golden gate or directly knocking TF33 out.
Proper citation: RRID:Addgene_219898 Copy
Species: Homo sapiens
Genetic Insert: von Hippel-Lindau tumor suppressor
Vector Backbone Description: Backbone Size:8400; Vector Backbone:pLEX_307 (Plasmid #41392); Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:38514884
Proper citation: RRID:Addgene_220147 Copy
Species: Homo sapiens
Genetic Insert: Bcl2
Vector Backbone Description: Backbone Size:4702; Vector Backbone:pEMD-C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Proper citation: RRID:Addgene_220301 Copy
Species: Homo sapiens
Genetic Insert: von Hippel-Lindau tumor suppressor
Vector Backbone Description: Backbone Size:8400; Vector Backbone:pLEX_307 (Plasmid #41392); Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:38514884
Proper citation: RRID:Addgene_220149 Copy
Species: Homo sapiens
Genetic Insert: Bcl2
Vector Backbone Description: Backbone Size:4702; Vector Backbone:pEMD-C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Proper citation: RRID:Addgene_220304 Copy
Species: Homo sapiens
Genetic Insert: Bcl2
Vector Backbone Description: Backbone Size:4702; Vector Backbone:pEMD-C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Proper citation: RRID:Addgene_220300 Copy
Vector Backbone Description: Backbone Size:4457; Vector Backbone:pERASE; Vector Types:Synthetic Biology; Bacterial Resistance:Erythromycin
Defining Citation: PMID:38911550
Comments: This plasmid allows to obtain scarless gene deletions in S. pyogenes and potentially other Gram-positive bacteria. The multiple cloning site (EcoRI, XbaI, SpeI, PstI) contains a Plac_mrfp cassette, allowing for red-white screening of clones in E. coli (DH5a). When the Plac_mrfp cassette is replaced with the insert of interest, colonies will appear white on LB agar plates. See the associated publication for more information.
Resource Information: The pUC origin for replication in E. coli was taken from pUC19 (Plasmid #50005). The erythromycin resistance cassette was taken from p7INT. The MCS containing the mrfp cassette was taken from pSpy0C4. The coding sequence for the counterselection marker pheS* was taken from the native pheS coding sequence found in S. pyogenes SF370. This sequence was adapted to avoid recombination events of the plasmid at the phis locus due to high DNA sequence homology. Two mutations were introduced into the adapted pheS sequence to facilitate incorporation of the toxic phenylalanine analogue PCPA (4-chlor-DL-phenylalanine) into proteins, leading to bacterial cell death.
p7INT was a gift from Prof. Michael Federle (University of Illinois Chicago, USA). The p7INT plasmid integrates into the 3' end of the tmRNA locus in several S. pyogenes strains. It was originally described in:
McShan, W.M., McLaughlin, R.E., Nordstrand, A. et al. Vectors containing streptococcal bacteriophage integrases for site-specific gene insertion. Methods Cell Sci 20, 51–57 (1998). https://doi.org/10.1023/A:1009773309163
Please download the detailed plasmid map using the file linked below.
Proper citation: RRID:Addgene_220306 Copy
Species: Escherichia coli
Genetic Insert: PhoB promoter only
Vector Backbone Description: Vector Backbone:pBAC (Addgene Plasmid #13423); Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:38687030
Comments: Please visit https://www.biorxiv.org/content/10.1101/2023.11.28.568972v1 for bioRxiv preprint.
Proper citation: RRID:Addgene_220163 Copy
Species: Escherichia coli
Genetic Insert: PhoP promoter only
Vector Backbone Description: Vector Backbone:pBAC (Addgene Plasmid #13423); Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:38687030
Comments: Please visit https://www.biorxiv.org/content/10.1101/2023.11.28.568972v1 for bioRxiv preprint.
Proper citation: RRID:Addgene_220164 Copy
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