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Species: Homo sapiens
Genetic Insert: Ataxin-1
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:8123; Vector Backbone:pDEST27; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:18337722
Proper citation: RRID:Addgene_21758 Copy
Species: Homo sapiens
Genetic Insert: Ataxin-1
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:8123; Vector Backbone:pDEST27; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:18337722
Proper citation: RRID:Addgene_21756 Copy
Vector Backbone Description: Backbone Size:3508; Vector Backbone:pMLM36; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:19798082
Comments: Combinatorial libraries of zinc finger arrays made by OPEN are cloned into this plasmid (see Maeder et al., Mol Cel 2008, PMID 18657511). Ligate zinc finger array fragments with BbsI-digested pBR-UV5-GP-FD2 vector backbone to create plasmids that express the zinc finger array as a FLAG-tagged Gal11P fusion in the B2H system.
The lacUV5 promoter which drives expression of the Gal11P fusion protein contains a lac operator and therefore can be repressed by lac repressor.
This plasmid is a low copy number plasmid and therefore we recommend that minipreps be performed from a 4 to 10 ml culture.
Annotations for pMLM36:
12-17: lacUV5 promoter -35 box / 36-41: lacUV5 promoter -10 box / 48: lacUV5 promoter transcription start point / 86-88: Start codon for Gal11P protein fragment / 89-358: Coding sequence for Gal11P a.a. 263-352 (with additional translationally silent mutations) / 407-412: BbsI site #1 / 433-438: BbsI site #2 / 568-1024: f1 phage origin of replication / 1156-2013: beta-lactamase gene cassette (confers resistance to ampicillin and carbenicillin) / 2016-2956: ColE1 origin of replication (plasmid origin) / 3203-3390: rop gene (associated with regulation of copy number)
Proper citation: RRID:Addgene_21871 Copy
Genetic Insert: CSH100 bacterial Strain
Vector Backbone Description: Backbone Size:0; Vector Backbone:n/a; Vector Types:; Bacterial Resistance:None
Defining Citation: PMID:19798082
Comments: F’ lac proA+proB+(lacIq lacPL8)/ara- ∆(gpt-lac)5
This strain will be shipped as bacteria in an LB stab. Upon receipt, requesting scientists should restreak the strain on an M9 minimal plate. After restreaking on M9 to confirm the presence of the F', scientists can grow the strain in liquid LB.
M9 minimal medium agar plates: To prepare 500 ml, autoclave 439 ml H2O with 7.5 g Bacto-agar and a stir bar. When agar has cooled to approximately 65°C, add 50 ml 10X M9 salts, 1 ml 1 M MgSO4, 10 ml 20% (w/v) glucose and 0.5 ml 100mM CaCl2 and then pour plates. Plates can be stored indefinitely at 4°C in sealed plastic bags. (Alternatively, M9 plates can be purchased from Teknonva: https://www.teknova.com/content/teknova/us/en/products/product-page.html/m1260.html).
Proper citation: RRID:Addgene_21875 Copy
Genetic Insert: KJ1C bacterial Strain
Vector Backbone Description: Backbone Size:0; Vector Backbone:n/a; Vector Types:; Bacterial Resistance:Tetracycline
Defining Citation: PMID:19798082
Comments: F- ∆hisB463 ∆(gpt-proAB-arg-lac)XIII zaj::Tn10
Proper citation: RRID:Addgene_21874 Copy
Vector Backbone Description: Backbone Size:5725; Vector Backbone:pMLM292; Vector Types:Mammalian Expression, zebrafish expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20010934
Comments: Zinc fingers are cloned in between the unique XbaI and BamHI sites. This results in an expression vector which encodes a ZFN with the OPEN zinc finger array fused to the mutant FokI nuclease domain.
pMLM292 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
235- 822: CMV promoter / 771-775: CAAT box / 804-808: TATA box / 819: 3' end of hCMV / 904-906: ZFN start codon / 910-930: SV40 nuclease localization signal (NLS) / 940- 963: FLAG tag / 964- 969: XbaI site / 980- 985: BamHI site / 986-1573: FokI nuclease domain / 1292&1294: Q486E heterodimer FokI mutation / 1331: I499L heterodimer FokI mutation / 1574-1576: ZFN stop codon / 1686-1910: BGH polyadenylation sequence / 1956-2384: f1 origin / 2411-2720: SV40 early promoter and origin / 2775-2841: EM-7 Promoter / 2842-3240: Blasticidin resistance gene / 3398-3528: SV40 late polyA signal / 3955-4574: pUC origin / 4729-5589: ampicillin (bla) resistance gene (complementary strand) / 5590-5688: bla promoter (complementary strand)
Proper citation: RRID:Addgene_21873 Copy
Species: Homo sapiens
Genetic Insert: SHP-1
Vector Backbone Description: Backbone Size:7767; Vector Backbone:pcDNA-Dest53; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17575265
Proper citation: RRID:Addgene_21802 Copy
Species: Mus musculus
Genetic Insert: PSMD14
Vector Backbone Description: Backbone Marker:Clontech (?); Backbone Size:4200; Vector Backbone:pEYFP; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:16973439
Comments: Note that though there is a Kan resistance gene in this plasmid according to the author's sequence, the gene is not functional. This plasmid does NOT confer Kan resistance.
Proper citation: RRID:Addgene_21801 Copy
Species: Mus musculus
Genetic Insert: PSMD14
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5446; Vector Backbone:pcDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:16973439
Proper citation: RRID:Addgene_21800 Copy
Genetic Insert: ECFP -EYFP linked by flexible linker containing 7 SGGSGG repeats
Vector Backbone Description: Backbone Marker:Novagen; Backbone Size:5200; Vector Backbone:pET28; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:17073440
Proper citation: RRID:Addgene_21766 Copy
Species: Homo sapiens
Genetic Insert: SHP-1
Vector Backbone Description: Backbone Size:7420; Vector Backbone:pDest472; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17575265
Proper citation: RRID:Addgene_21806 Copy
Species: Homo sapiens
Genetic Insert: SHP-1
Vector Backbone Description: Backbone Size:7420; Vector Backbone:pDest472; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17575265
Proper citation: RRID:Addgene_21804 Copy
Genetic Insert: ECFP -EYFP linked by flexible linker containing two SGGSGG repeats
Vector Backbone Description: Backbone Marker:Novagen; Backbone Size:5200; Vector Backbone:pET28; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:17073440
Proper citation: RRID:Addgene_21761 Copy
Genetic Insert: ECFP -EYFP linked by flexible one SGGSGG repeats
Vector Backbone Description: Backbone Marker:Novagen; Backbone Size:5200; Vector Backbone:pET28; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:17073440
Proper citation: RRID:Addgene_21760 Copy
Species: Mus musculus
Genetic Insert: IRE1b
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5446; Vector Backbone:pcDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Comments: Mouse IRE1b, wild-type, 9E10-tagged at C-term, assembled from lambda-phage clones 37 and 26, in pCDNA3.
Proper citation: RRID:Addgene_21880 Copy
Genetic Insert: ECFP -EYFP linked by flexible linker containing 4 SGGSGG repeats
Vector Backbone Description: Backbone Marker:Novagen; Backbone Size:5200; Vector Backbone:pET28; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:17073440
Proper citation: RRID:Addgene_21763 Copy
Genetic Insert: ECFP -EYFP linked by flexible linker containing 3 SGGSGG repeats
Vector Backbone Description: Backbone Marker:Novagen; Backbone Size:5200; Vector Backbone:pET28; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:17073440
Proper citation: RRID:Addgene_21762 Copy
Species: Mus musculus
Genetic Insert: ATF4
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4733; Vector Backbone:pEGFP-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:15479734
Comments: These pEGFP-based plasmids have NEOr marker (and not the conventional AMPr marker). They should be propagated in the presence of 30mcg/ml kanamycin (and not on ampicillin)
Proper citation: RRID:Addgene_21857 Copy
Species: Mus musculus
Genetic Insert: ATF4
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4733; Vector Backbone:pEGFP-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:15479734
Comments: These pEGFP-based plasmids have NEOr marker (and not the conventional AMPr marker). They should be propagated in the presence of 30mcg/ml kanamycin (and not on ampicillin)
Proper citation: RRID:Addgene_21856 Copy
Species: Mus musculus
Genetic Insert: ATF4
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4733; Vector Backbone:pEGFP-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:15479734
Comments: These pEGFP-based plasmids have NEOr marker (and not the conventional AMPr marker). They should be propagated in the presence of 30mcg/ml kanamycin (and not on ampicillin)
Proper citation: RRID:Addgene_21855 Copy
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