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Species: Mus musculus
Genetic Insert: PKC eta
Vector Backbone Description: Backbone Size:5400; Vector Backbone:pHACE; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments: Osada, S., Mizuno, K., Saido, T.C., Akita, Y., Suzuki, K., Kuroki, T., and Ohno, S. 1990. A phorbol ester receptor/protein kinase, nPKC eta, a new member of the protein kinase C family predominantly expressed in lung and skin. The Journal of biological chemistry 265(36): 22434-22440.
Proper citation: RRID:Addgene_21245 Copy
Species: Mus musculus
Genetic Insert: PKC epsilon
Vector Backbone Description: Backbone Size:5400; Vector Backbone:pHACE; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_21240 Copy
Species: Rattus norvegicus
Genetic Insert: GFP-N2-PKCdelta-C1
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4596; Vector Backbone:N2; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
References:
Comments:
Proper citation: RRID:Addgene_21216 Copy
Species: Homo sapiens
Genetic Insert: Mek2
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:2200; Vector Backbone:pENTR1A; Vector Types:Subclone vector; Bacterial Resistance:Kanamycin
References:
Comments: Constitutive active protein has the following aa sequence deleted - EQQKKRLE, residues 48 to 55.
Proper citation: RRID:Addgene_21211 Copy
Species: Homo sapiens
Genetic Insert: Phosphoglycerate kinase promoter
Vector Backbone Description: Backbone Size:7226; Vector Backbone:SIN18; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
References:
Comments: There are several mismatches between depositor's and Addgene sequence. These mutations do not affect eGFP expression.
Proper citation: RRID:Addgene_21210 Copy
Species: Homo sapiens
Genetic Insert: DUX4
Vector Backbone Description: Backbone Size:4095; Vector Backbone:pCS2; Vector Types:Mammalian Expression, Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_21176 Copy
Species: Rattus norvegicus
Genetic Insert: GFP-pcDNA3-PKCgamma-c2
Vector Backbone Description: Backbone Size:5429; Vector Backbone:pcDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_21215 Copy
Species: Homo sapiens
Genetic Insert: JMJD3
Vector Backbone Description: Backbone Size:6300; Vector Backbone:pMSCVpuro; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_21214 Copy
Species:
Genetic Insert: PM-GFP
Vector Backbone Description: Backbone Size:5419; Vector Backbone:pcDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_21213 Copy
Species: Homo sapiens
Genetic Insert: PLCdelta
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4700; Vector Backbone:C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
References:
Comments: This construct encodes a PI(4,5)P2 lipid selective PH domain that can be used as a fluorescent translocation biosensor to monitor changes or local differences in the concentration of plasma membrane PI(4,5)P2 lipids (Stauffer TP, Ahn S, Meyer T; Curr Biol. 1998 8:343-6). It is best used in combination with a second fluorescent protein that uniformly marks the plasma membrane (the last 20 amino acids "CAAX" region of KRas is often used as a PM marker). Relative changes in cell-wide or local plasma membrane PI(4,5)P2 can be determined by using a mask for the PM region and by dividing the two background subtracted PH domain and CAAX images. At high expression levels, this construct interferes with PI(4,5)P2 signaling and alters cortical actin (Raucher D et al Cell. 2000 100:221-8).
Proper citation: RRID:Addgene_21179 Copy
Species: Homo sapiens
Genetic Insert: JMJD3
Vector Backbone Description: Backbone Size:6300; Vector Backbone:pMSCVpuro; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_21212 Copy
Species: Homo sapiens
Genetic Insert: E2F transcription factor 1
Vector Backbone Description: Backbone Marker:Promega; Backbone Size:5256; Vector Backbone:pGL3-Control; Vector Types:Mammalian Expression, Luciferase; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_21170 Copy
Species: Homo sapiens
Genetic Insert: DUX4
Vector Backbone Description: Backbone Size:4095; Vector Backbone:pCS2; Vector Types:Mammalian Expression, Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_21172 Copy
Species: Homo sapiens
Genetic Insert: E2F 3'UTR Mutant
Vector Backbone Description: Backbone Marker:Promega; Backbone Size:5256; Vector Backbone:pGL3-Control; Vector Types:Mammalian Expression, Luciferase; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_21171 Copy
Species: Mus musculus
Genetic Insert: alpha myosin heavy chain promoter
Vector Backbone Description: Backbone Size:8400; Vector Backbone:SIN18; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_21229 Copy
Species: Mus musculus
Genetic Insert: alpha myosin heavy chain promoter
Vector Backbone Description: Backbone Size:7338; Vector Backbone:SMPU; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
References:
Comments: There is an extra C in the promoter sequence when compared to the available reference sequence in NCBI. The mismatch is due to older sequencing artifact of an erroneous insertion of a "C" adjacent to "GGG". The construct should be correct.
Proper citation: RRID:Addgene_21228 Copy
Species:
Genetic Insert: GateWay conversion cassette
Vector Backbone Description: Backbone Size:14820; Vector Backbone:pRosa26-PA; Vector Types:Mammalian Expression; Bacterial Resistance:Chloramphenicol and Ampicillin
References:
Comments: pRosa26-DEST was made by inserting a GateWay conversion cassette into the XhoI site of pBigT, followed by transfer of PacI / AscI fragment of the resulting vector to the pRosa26-PA vector (see http://www.biomedcentral.com/1471-213X/1/4
for details on these vectors). The resulting vector resembles the construct that was used for the R26R Cre reporter mouse made by Phil Soriano in which the expression of lacZ is coupled to the endogenous Rosa26 locus via a splice acceptor site, but only after removal of the lox-STOP-lox cassette (which is actually the neo-R cassette used during the targeting). The only thing changed from the pBigT vectors and the R26R reporter is the cloning strategy, so the resulting knock-in should be as good as these systems, and as good (or as bad) as the use of the Rosa26 locus in any situation.
Stbl3 cells are advisable to use after users have done the LR recombination reaction.
See http://www.hgu.mrc.ac.uk/people/n.hastie_rosa.html
for more information and protocols for this plasmid.
Further advice from the depositing lab:
"Grow single colonies in Amp + Cam at 30 C, definitively not 37. Including the Cam should ensure that spontaneous recombinants that loose the Gateway cassette don't survive...
- Do minipreps and test with the double digest PacI / AscI. This should give 4.9 kb + 9.8 kb. If this is okay, the vector should be okay"
Please note that there are discrepancies between Addgene's quality control sequence and the theoretical sequence supplied by the depositing lab. These discrepancies should not affect the function of the construct.
Proper citation: RRID:Addgene_21189 Copy
Species: Rattus norvegicus
Genetic Insert: CAMKIIalpha-T286A
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:0; Vector Backbone:C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
References:
Comments:
Proper citation: RRID:Addgene_21220 Copy
Species: Rattus norvegicus
Genetic Insert: CAMKIIalpha
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4725; Vector Backbone:C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
References:
Comments:
Proper citation: RRID:Addgene_21226 Copy
Species: Rattus norvegicus
Genetic Insert: CaMKIIbeta(A303R)
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:0; Vector Backbone:C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
References:
Comments:
Proper citation: RRID:Addgene_21224 Copy
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