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Species: Homo sapiens
Genetic Insert: Cre recombinase
Vector Backbone Description: Backbone Marker:Michael Brenner (PMID: 7952266); Backbone Size:5500; Vector Backbone:pgfa2-lac2; Vector Types:Mammalian Expression, Mouse Targeting, Cre/Lox; Bacterial Resistance:Ampicillin
Defining Citation: PMID:11668683
Comments: This plasmid encodes a transgene to express the Cre recombinase in astrocytes of transgenic mice. A DNA fragment encoding the Cre recombinase was inserted into an expression cassette containing the gfa2 promoter (Besnard et al., 1991), a 2.2-kb 5' flanking region from the human GFAP (hGFAP) gene. The nuclear targeting signal from the SV40 large T antigen was inserted at the amino terminal end of the coding region, because it reportedly increases the efficiency of DNA excision by the recombinase (Gu et al., 1993). A heterologous intron and polyadenylation signal were provided by sequences from the mouse protamine 1 gene at the 3' end.
The hGFAP-cre transgene was constructed by excising the lacZ coding region from pgfa2-lac2 (Brenner et al., 1994) by BamHI digestion, and replacing it by blunt end ligation with a nuclear-targeted Cre coding sequence. The latter was obtained as a 1.1-kb SalI-MluI fragment from plasmid pTZ19RCreNLS, which was kindly provided by Drs. Raul Torres and Klaus Rajewsky. Both junctions were verified by DNA sequencing.
Proper citation: RRID:Addgene_40591 Copy
Species: Homo sapiens
Genetic Insert: PLXNA4A
Vector Backbone Description: Backbone Marker:SGC; Vector Backbone:pET28-MHL; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Comments: http://www.thesgc.org/structures/4E74/
Proper citation: RRID:Addgene_40748 Copy
Species: Mus musculus
Genetic Insert: Syndecan-1 shRNA(UTR)
Vector Backbone Description: Backbone Size:10130; Vector Backbone:FUGW-H1; Vector Types:Lentiviral, RNAi; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22936997
Comments: This shRNA plasmid was generated by inserting the hairpin oligonucleotides into the FUGW-H1
lentiviral construct.
Proper citation: RRID:Addgene_40623 Copy
Species: Homo sapiens
Genetic Insert: Wnt reporter sequences
Vector Backbone Description: Backbone Marker:addgene; Vector Backbone:pSicoR PGK Puro; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22177620
Comments: This lentiviral Wnt reporter has 7 repeats of Tcf/Lef binding sites, which was cloned by Xiaojun (Lance) Lian.
Methods for Plasmid construction, lentiviral production and infection of hESCs:
The 7 TCF/LEF binding site sequences were obtained by direct PCR of plasmid M50 (Addgene plasmid 12456) and the GreenFire (GF) sequences were obtained by direct PCR of the pGreenFire1-mCMV Plasmid (System Bioscience TR010PA-1). These two sequences were cloned into pSicoR PGK puro (Addgene plasmid 11586) digested by Xba I (NEB) and Xho I (NEB). The constructed Wnt reporter plasmid was named 7TGFP and verified by sequencing. This 7TGFP vector was cotransfected with the helper plasmids psPAX2 and pMD2.G (Addgene plasmids 12260 and 12259) into HEK-293TN cells (System Biosciences) for virus production. Virus-containing medium was collected at 48 and 72 hours after transfection and used for infection of hESCs in the presence of 6μg/mL polybrene (Sigma). Transduced cells were cultured in mTeSR1 on Matrigel for three days and then clonally isolated in mTeSR1 with 1 μg/mL puromycin.
Proper citation: RRID:Addgene_40588 Copy
Species: Homo sapiens
Genetic Insert: TCF7
Vector Backbone Description: Backbone Size:5400; Vector Backbone:pcDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22949634
Proper citation: RRID:Addgene_40620 Copy
Species: Trypanosoma Brucei
Genetic Insert: Trypanosoma Brucei Adenylosuccinate Lyase
Vector Backbone Description: Backbone Marker:SGC; Vector Backbone:pET15-MHL; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: http://www.thesgc.org/structures/4EFC/
Tb427tmp.160.5560:MAC049-C01:C223709
Proper citation: RRID:Addgene_40749 Copy
Species: Arabidopsis thaliana
Genetic Insert: VP16-CIB1
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:7500; Vector Backbone:pGADT7rec; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22922268
Proper citation: RRID:Addgene_40615 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: HIS3
Vector Backbone Description: Backbone Marker:Miller et al (1998) Nucleic Acids Res 26(15):3577-3583 ; Backbone Size:6512; Vector Backbone:pBEVY-U; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24357599
Proper citation: RRID:Addgene_40610 Copy
Species: Synthetic
Genetic Insert: Aurora B FRET-sensor
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:5125; Vector Backbone:pIRES-puro2b; Vector Types:Mammalian Expression, Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22801782
Proper citation: RRID:Addgene_40731 Copy
Species: Synthetic
Genetic Insert: Leucine Zipper prime - C super positive Green Fluorescent Protein
Vector Backbone Description: Backbone Size:4496; Vector Backbone:pMRBAD; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:22692102
Proper citation: RRID:Addgene_40730 Copy
Species: Homo sapiens
Genetic Insert: PA1
Vector Backbone Description: Backbone Marker:System Biosciences (SBI); Backbone Size:8295; Vector Backbone:pCDH-EF1-MCS-IRES-Neo; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22949634
Proper citation: RRID:Addgene_40618 Copy
Species: Homo sapiens
Genetic Insert: RPRD1B
Vector Backbone Description: Backbone Marker:SGC; Vector Backbone:pET15-MHL; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: http://www.thesgc.org/structures/4FLA/
Proper citation: RRID:Addgene_40739 Copy
Species: Homo sapiens
Genetic Insert: L3MBTL3
Vector Backbone Description: Backbone Marker:SGC; Vector Backbone:pNIC-CH; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Comments: http://www.thesgc.org/structures/4FL6/
Proper citation: RRID:Addgene_40738 Copy
Species: Mus musculus
Genetic Insert: AMPK-γ1
Vector Backbone Description: Backbone Size:4300; Vector Backbone:pCMV-Tag4A; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:17012231
Comments: AMPK-γ1 insert has Kozak sequence embedded at 5'-terminus and a stop codon at the 3'-terminus. The FLAG of the pCMV-Tag4a backbone is out of frame.
Proper citation: RRID:Addgene_40605 Copy
Species: Mus musculus
Genetic Insert: AMPK-β2
Vector Backbone Description: Backbone Marker:Stratagene; Backbone Size:4300; Vector Backbone:pCMV-Tag4A; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:17012231
Proper citation: RRID:Addgene_40603 Copy
Species: Mus musculus
Genetic Insert: AMPK-β1
Vector Backbone Description: Backbone Size:4300; Vector Backbone:pCMV-Tag4A; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:17012231
Proper citation: RRID:Addgene_40602 Copy
Species: Photinus pyralis
Genetic Insert: Firefly Luciferase
Vector Backbone Description: Backbone Marker:Miller et al (1998) Nucleic Acids Res 26(15):3577-3583 ; Backbone Size:6518; Vector Backbone:pBEVY-U; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24357599
Proper citation: RRID:Addgene_40601 Copy
Species: Homo sapiens
Genetic Insert: LRRK2
Vector Backbone Description: Backbone Marker:Emerald; Vector Backbone:pEMB108; Vector Types:Baculovirus; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_40567 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: Renilla Luciferase
Vector Backbone Description: Backbone Marker:Miller et al (1998) Nucleic Acids Res 26(15):3577-3583 ; Backbone Size:6518; Vector Backbone:pBEVY-U; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24357599
Proper citation: RRID:Addgene_40600 Copy
Species: Homo sapiens
Genetic Insert: LRRK2
Vector Backbone Description: Backbone Marker:Emerald; Vector Backbone:pEMB52; Vector Types:Baculovirus; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_40565 Copy
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