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Species: Bos taurus
Genetic Insert: Bovine 70 kDa heat shock cognate protein fragment amino acids 1-554
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:2950; Vector Backbone:pRSET_A; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_12533 Copy
Species: Bos taurus
Genetic Insert: bovine 70 kDa heat shock cognate protein full length wild type
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:2950; Vector Backbone:pRSET_A; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_12532 Copy
Species: Bos taurus
Genetic Insert: p110-CAAX
Vector Backbone Description: Backbone Size:5558; Vector Backbone:pBabe hygro; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
References:
Comments: Myc tagged p110-CAAX was cut out of pBabe-puro p110-CAAX using BamHI and ligated into BamHI cut pBabe-hygro.
Proper citation: RRID:Addgene_12591 Copy
Species: Bos taurus
Genetic Insert: gRNA-GFPDROPOUT-for Sp Cas9 bu6_20N_cr2
Vector Backbone Description: Vector Backbone:MTK0_027; Vector Types:Mammalian Expression; Bacterial Resistance:Chloramphenicol
References:
Comments: Type 234 parts are coupled promoter-coding sequence-terminator such as guide RNA cassettes, GFP dropouts in the MTK Please visit https://www.biorxiv.org/content/10.1101/506188v2 for BioRxiv preprint
Proper citation: RRID:Addgene_123920 Copy
Species: Bos taurus
Genetic Insert: bovine GRK2-CAAX
Vector Backbone Description: Vector Backbone:pRK5; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_166224 Copy
Species: Bos taurus
Genetic Insert: Rap1b N17
Vector Backbone Description: Backbone Marker:invitrogen; Vector Backbone:pcDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments: site directed mutagenesis
Proper citation: RRID:Addgene_118322 Copy
Species: Bos taurus
Genetic Insert: PKC alpha
Vector Backbone Description: Backbone Size:4650; Vector Backbone:pCMV6; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments: Constitutively active. Myr sequence from p60src.
Proper citation: RRID:Addgene_10807 Copy
Species: Bos taurus
Genetic Insert: PKC alpha
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5446; Vector Backbone:pcDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_10806 Copy
Species: Bos taurus
Genetic Insert: p110 KD
Vector Backbone Description: Backbone Marker:BD Biosciences; Backbone Size:6620; Vector Backbone:pLNCX; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
References:
Comments: A pair of overlapping oligomers were synthesized based on the c-src myristoylation signal peptide sequence. A Kozak sequence was engineered at the 5 of the c-src myristoylation signal sequence (MetGly-Ser-Ser-Lys-Ser-Lys-Pro-Lys-Asp-Pro-Ser-Gln-Arg-Arg-ArgArg-Ile-Arg-Thr). The fragment was ligated to the 5 end of bovine p110 sequence and cloned into a retroviral expression vector (pLNCX) by a three piece ligation reaction. A 6 histidine tag was incorporated on the 3 end of p110 by PCR. The kinase dead mutant has an additional BamHI site at D915S.
Proper citation: RRID:Addgene_10837 Copy
Species: Bos taurus
Genetic Insert: p110
Vector Backbone Description: Backbone Marker:BD Biosciences; Backbone Size:6620; Vector Backbone:pLNCX; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
References:
Comments: A pair of overlapping oligomers were synthesized based on the c-src myristoylation signal peptide sequence. A Kozak sequence was engineered at the 5 of the c-src myristoylation signal sequence (MetGly-Ser-Ser-Lys-Ser-Lys-Pro-Lys-Asp-Pro-Ser-Gln-Arg-Arg-ArgArg-Ile-Arg-Thr). The fragment was ligated to the 5 end of bovine p110 sequence and cloned into a retroviral expression vector (pLNCX) by a three piece ligation reaction. A 6 histidine tag was incorporated on the 3 end of p110 by PCR.
Proper citation: RRID:Addgene_10836 Copy
Species: Bos taurus
Genetic Insert: p110alpha
Vector Backbone Description: Backbone Size:5100; Vector Backbone:pWZL Blast; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_10889 Copy
Species: Bos taurus
Genetic Insert: 14-3-3 zeta
Vector Backbone Description: Backbone Marker:Amersham; Backbone Size:4969; Vector Backbone:pGEX-4T1; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments: Insert PCR'd and cloned into pGEX-4T1 (via either EcoRI and/or BamHI?) by Tom Roberts lab.
Uses stop codons of pGEX backbone rather than 14-3-3 and contains a small extra C-terminal extension. This has been corrected in the HA-tagged constructs.
Proper citation: RRID:Addgene_13278 Copy
Species: Bos taurus
Genetic Insert: pI3K p110 CAAX
Vector Backbone Description: Backbone Size:5169; Vector Backbone:pBabe puro; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
References:
Comments: Plasma membrane-targeted, constitutively activated PI3K p110. Please note that the myc tag is NONFUNCTIONAL.
Proper citation: RRID:Addgene_13339 Copy
Species: Bos taurus
Genetic Insert: BCA-Q11
Vector Backbone Description: Vector Backbone:pET28a; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
References:
Comments:
Proper citation: RRID:Addgene_185781 Copy
Species: Bos taurus
Genetic Insert: BCA-P(11)4
Vector Backbone Description: Vector Backbone:pET28a; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
References:
Comments:
Proper citation: RRID:Addgene_185780 Copy
Species: Bos taurus
Genetic Insert: Bovine GRK2[D110A K220R]
Vector Backbone Description: Vector Backbone:pEGFP-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
References:
Comments:
Proper citation: RRID:Addgene_137779 Copy
Species: Bos taurus
Genetic Insert: PI3K catalytic subunit p110-alpha
Vector Backbone Description: Backbone Size:5000; Vector Backbone:pCDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_1397 Copy
Species: Bos taurus
Genetic Insert: PIK3CA E545K
Vector Backbone Description: Backbone Marker:Jeff Engelman; Backbone Size:0; Vector Backbone:JP1520; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
References:
Comments: Hemagglutinin (HA)–tagged bovine PIK3CA cDNA was subcloned into the pDNR-Dual vector (Clontech, Mountain View, CA), and site-directed mutagenesis (Quick Change, Stratagene, La Jolla, CA) was used to generate the E545K mutation. The cDNA was shuttled into the JP1520 retroviral vector using the BD Creator System (Clontech).
Proper citation: RRID:Addgene_14571 Copy
Species: Bos taurus
Genetic Insert: BARK1 minigene
Vector Backbone Description: Backbone Size:4700; Vector Backbone:pRK5; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments: The cDNA encoding bovine BARK1 residues Gly495 to Leu689 was amplified using PCR to construct a minigene casette with newly engineered 5' and 3' ends (see Author's map). The 5' end contained an EcoRI site for subcloning, followed by the kozak sequence 5'-GCCGCCACCATGG-3'. A bclI site was inserted at the 3' end immediately following the stop codon. The final BARK1 minigene construct contains the 3'UTR of the human beta-globin gene and a polyadenylation signal.
Proper citation: RRID:Addgene_14695 Copy
Species: Bos taurus
Genetic Insert: PKA R1alpha regulatory subunit
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:2900; Vector Backbone:pRSET B; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_14922 Copy
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