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Species: Homo sapiens
Genetic Insert: API5 apoptosis inhibitor 5
Vector Backbone Description: Backbone Size:5496; Vector Backbone:pFRT/TO/HIS/FLAG/HA-DEST; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22681889
Proper citation: RRID:Addgene_38074 Copy
Species: Homo sapiens
Genetic Insert: KIAA1967
Vector Backbone Description: Backbone Size:5539; Vector Backbone:pFRT/TO/HIS/FLAG/HA-DEST; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22681889
Proper citation: RRID:Addgene_38082 Copy
Species: Homo sapiens
Genetic Insert: MYBBP1A MYB binding protein (P160) 1a
Vector Backbone Description: Backbone Size:5499; Vector Backbone:pFRT/TO/HIS/FLAG/HA-DEST; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22681889
Proper citation: RRID:Addgene_38084 Copy
Species: Homo sapiens
Genetic Insert: HNRNPR heterogeneous nuclear ribonucleoprotein R
Vector Backbone Description: Backbone Size:5538; Vector Backbone:pFRT/TO/HIS/FLAG/HA-DEST; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22681889
Proper citation: RRID:Addgene_38067 Copy
Species: Homo sapiens
Genetic Insert: HNRNPU heterogeneous nuclear ribonucleoprotein U (scaffold attachment factor A)
Vector Backbone Description: Backbone Size:5496; Vector Backbone:pFRT/TO/HIS/FLAG/HA-DEST; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22681889
Proper citation: RRID:Addgene_38068 Copy
Species: Homo sapiens
Genetic Insert: MYEF2 myelin expression factor 2
Vector Backbone Description: Backbone Size:5508; Vector Backbone:pFRT/TO/HIS/FLAG/HA-DEST; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22681889
Comments: Digest and NGS results suggest that the original Gateway entry vector is still present in the DNA stock from which the bacterial stocks are created. Because the entry vector does not contain a promoter for the insert or selection marker, it should not interfere with function.
To attempt to remove the entry vector, recipient scientists could try to cure the plasmid from isolated colonies. Alternatively, they can linearize the entry vector with BmtI, HpaI or NheI digest, which will not cut the expression vector, and retransform into bacteria.
Proper citation: RRID:Addgene_38069 Copy
Species: Homo sapiens
Genetic Insert: HNRNPD heterogeneous nuclear ribonucleoprotein D (AU-rich element RNA binding protein 1, 37kDa)
Vector Backbone Description: Backbone Size:5496; Vector Backbone:pFRT/TO/HIS/FLAG/HA-DEST; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22681889
Proper citation: RRID:Addgene_38066 Copy
Species: Homo sapiens
Genetic Insert: BCR/ABL P190
Vector Backbone Description: Backbone Size:7400; Vector Backbone:pLEF; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:18070886
Comments: Constructed by Suparna Mishra. Generated in a 3-way ligation consisting of 5' 0.75 kb BamHI/Sal I + 3' 6.3 kb Sal I/EcoRI BCR/ABL fragments into pLEF digested with BamHIxEcoRI. The ABL part of the insert contains the natural TAG and 3' UT sequences.
Addgene NGS identified a sequence discrepancy at nucleotide 3168 resulting in a T117M mutation in the ABL-1 sequence, relative to Genbank ID NP_005148.2 and Addgene Plasmid #38158. This is not a known polymorphism and the effect of this, if any, on the protein is not known.
Proper citation: RRID:Addgene_38159 Copy
Species: Homo sapiens
Genetic Insert: Abr
Vector Backbone Description: Backbone Marker:Don Kohn [email protected]; Backbone Size:6600; Vector Backbone:pCCL-cppt178-MNDU3; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17116687
Comments: Constructed by Jess Cunnick. Abr was first subcloned into pEYFP-C1 Bgl II x KpnI as a 0.4 kb 5' BamHI/BstEII + 2.2 kb BstEII- KpnI fragment. The EYFP-Abr fusion was isolated as a 5' AgeI- 3' KpnI fragment and inserted into pCCL-cppt178-MNDU3-X3 digested with Xma I x Kpn I.
Proper citation: RRID:Addgene_38155 Copy
Species: Homo sapiens
Genetic Insert: ABR
Vector Backbone Description: Backbone Size:7400; Vector Backbone:pLEF; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:18070886
Comments: Constructed by Suparna Mishra. Generated in a 3-way ligation consisting of 5' 0.4 kb BamHI/BstEII + 3' 2.2 kb BstEII/EcoRI ABR fragments into pLEF digested with BamHIxEcoRI. The ABR insert contains the natural TAG and 3' UT sequences.
Addgene sequencing found an E24G point mutation--this does not alter plasmid function.
Proper citation: RRID:Addgene_38157 Copy
Species: Homo sapiens
Genetic Insert: Bcr
Vector Backbone Description: Backbone Size:7400; Vector Backbone:pLEF; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:14614449
Comments: Constructed by Anja Reichert. Generated in a 3-way ligation consisting of 5' 0.6 kb BamHI/SalI + 3' 3.5 kb SalI/XbaI BCR fragments into pLEF digested with BamHIxXbaI. The BCR insert contains the natural TAG and 3' UT sequences. At the 3' end there are polylinker sequences from pSK.
Proper citation: RRID:Addgene_38160 Copy
Species: Homo sapiens
Genetic Insert: Bcr/Abl P210
Vector Backbone Description: Backbone Marker:Pharmingen; Backbone Size:8530; Vector Backbone:pAcG2T; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Comments: Constructed by Nora Heisterkamp. Made using a 3-way ligation with 0.6 kb 5' BamHI-SalI from BCR + 3' 7.5 kb Sal I-EcoRI into pAcG2T. Not evaluated for protein production.
Proper citation: RRID:Addgene_38161 Copy
Species: Homo sapiens
Genetic Insert: MITF-A
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4733; Vector Backbone:pEGFP-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:22692423
Proper citation: RRID:Addgene_38132 Copy
Species: Homo sapiens
Genetic Insert: cIAP1 H588A
Vector Backbone Description: Backbone Marker:Baltimore Lab; Backbone Size:5350; Vector Backbone:pEBB; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:19243308
Proper citation: RRID:Addgene_38233 Copy
Species: Homo sapiens
Genetic Insert: BCR
Vector Backbone Description: Backbone Size:4805; Vector Backbone:pKH3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Comments: Constructed by Young Jin Cho.
A human BCR cDNA was inserted into pKH3 (Addgene plasmid #12555; obtained from Martin Schwartz) digested with BamHI x EcoRI in a three-way ligation (1.7 kb BamHI-SstI + 2.6 kb SstI-EcoRI). The 1.7 kb BamHI-SstI fragment was isolated from Bcr in pAcG2T (AddGene plasmid #38163). The 5’ sequence is
GGA TCC CCC GGG CTG CAG GAA TTC GAT GGC CGC GCC ATG-start BCR.
Proper citation: RRID:Addgene_38189 Copy
Species: Homo sapiens
Genetic Insert: BCR/ABL P190
Vector Backbone Description: Backbone Size:3000; Vector Backbone:pSK; Vector Types:construct for making transgenic mice; Bacterial Resistance:Ampicillin
Defining Citation: PMID:2179728
Comments: The DNA construct was made by Guido Jenster and Nora Heisterkamp in several cloning and subcloning steps. The transgenic construct contains human BCR exon 1, intron sequences of human BCR and ABL genes, human ABL exon 2 and ABL cDNA sequences. The transgene expression is controlled by a 200-bp Sst I- Bgl II fragment of the murine metallothionein-1 (Mt1) promoter lacking glucocorticoid-responsive elements (PMID 8541551). This BCR/ABL fusion gene produces a P190 protein and is a model for Philadelphia chromosome-positive acute lymphoblastic leukemia (ALL) in humans, which is caused by a chromosomal translocation that fuses the ABL proto-oncogene with breakpoint cluster region (BCR) sequences. Leukemia development is relatively rapid on inbred C57Bl/6J and CBA backgrounds and is slower on mixed genetic backgrounds. Mice transgenic for this construct on an inbred C57Bl/6J background are also available from Jackson labs stock 017833. The insert can be separated from the vector by digestion with Sst II.
Proper citation: RRID:Addgene_38185 Copy
Species: Homo sapiens
Genetic Insert: CRKL
Vector Backbone Description: Backbone Size:3500; Vector Backbone:pSL1180; Vector Types:mouse transgenic; Bacterial Resistance:Ampicillin
Defining Citation: PMID:11245441
Comments: This construct was made by Arnoud van Wijk. It can be used to make transgenic mice. The DNA construct contains 7.5 kb of human CRKL locus 5’ sequences including the promoter of the human v-crk sarcoma virus CT10 oncogene homolog (CRKL) gene, joined to CRKL exon 1. Part of the intron between exons 1 and 2 encompassing around 8-kb is included and exons 2-3 as cDNA. The entire insert is around 17 kb and can be isolated free from plasmid sequences in a Kpn I x Mlu I digestion. Jackson labs stock no. 017834 is one of our CRKL transgenics containing approximately 5 copies of this transgene. CRKL overexpressing transgenics are viable but mice with high copy numbers have reduced fertility.
How the CRKL transgenic construct was made: (verbatim from PMID 11245441): Human genomic clone 70 (PMID 7905853) contains 12 kb of 5’ sequences, exon 1, and 6.6 kb of intron 1 of the CRKL gene. A 350-bp RsaI fragment isolated from a CRKL cDNA (GenBank accession no. X59656) was used as a probe to specifically isolate a phage clone, CR-4, containing CRKL exon 2 flanked by 6.5 kb of intron 1 and 9.5 kb of intron 2. CRKL is located on human chromosome 22 and has been entirely sequenced; introns 1 and 2 are both around 15.5 kb. To generate a transgenic DNA construct, a 7-kb SalI-BamHI fragment from CR-4, which included a SalI site at the 5’ end from the phage polylinker and the 3’ BamHI site located in exon 2, was ligated with a 0.78-kb BamHI-EcoRI cDNA fragment, including exons 2, 3, and the 3’ untranslated region, into pSK digested with SalIxEcoRI. The insert was removed as a 7.7-kb SalI-NotI fragment and ligated with a 1.2-kb SstII-SalI fragment from intron 1 in clone 70 into pSK digested with SstII x NotI. The resulting insert was removed by digestion with SstII x NotI. The 5’ promoter and exon 1 sequences were isolated on an 8-kb EcoRI-SstII fragment that was subcloned into pSK digested with EcoRI x SstII; the insert was removed as an 8-kb KpnI-SstII fragment. The 8-kb KpnI-SstII fragment plus the 7.7-kb SstII-NotI fragment were ligated into pSL1180 digested with KpnI x NotI.
Proper citation: RRID:Addgene_38186 Copy
Species: Homo sapiens
Genetic Insert: autophagy-related genes 13
Vector Backbone Description: Backbone Marker:Dr. Toshio Kitamura of the University of Tokyo ; Backbone Size:5847; Vector Backbone:pMXs-IP; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:19211835
Proper citation: RRID:Addgene_38191 Copy
Species: Homo sapiens
Genetic Insert: ABR
Vector Backbone Description: Backbone Size:4805; Vector Backbone:pKH3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Comments: Constructed by Young Jin Cho.
A human ABR cDNA was inserted into pKH3 (Addgene plasmid 12555; obtained from Martin Schwartz) digested with BamHI x EcoRI in a three-way ligation (0.4 kb BamHI-BstEII + 2.2 kb BstEII-EcoRI). The 5’ sequence is
GGA TCC ATG-start ABR. Note: This plasmid may contain an E24G mutation -gag to ggg- introduced by Taq polymerase at an earlier cloning step.
Proper citation: RRID:Addgene_38190 Copy
Species: Homo sapiens
Genetic Insert: Abr GAP domain
Vector Backbone Description: Backbone Size:5500; Vector Backbone:pcDNA3.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17116687
Comments: Constructed by Young Jin Cho.
An 0.8 kb BamHI xEcoRI fragment from AbrGAP in pGEX3x (AddGene #36407) was inserted into pcDNA BamHI xEcoRI. The insert starts at a natural PvuII site in the ABR sequence. The fragment contains the natural TAG and 3’ UT sequences and encodes amino acid residues 597-859 (Abr isoform B).
Proper citation: RRID:Addgene_38177 Copy
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