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Species: Synthetic
Genetic Insert: GoldyTALEN
Vector Backbone Description: Backbone Size:4000; Vector Backbone:pBSK; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:23027955
Comments: For further reference, this plasmid was also used in the Bedell et al Nature 2012 publication (PMID 23000899).
For more information on Carlson TALEN Add-On Plasmids please refer to: http://www.addgene.org/TALeffector/goldengate/add-ons/#carlson
Proper citation: RRID:Addgene_38143 Copy
Species: M. anisopliae
Genetic Insert: Carboxypeptidase A
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:6526; Vector Backbone:pFastBac-1; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21073956
Proper citation: RRID:Addgene_38150 Copy
Species: Rattus norvegicus
Genetic Insert: 4EBP1
Vector Backbone Description: Backbone Marker:Broad Insitute; Backbone Size:5000; Vector Backbone:pCW57.1; Vector Types:Mammalian Expression, Lentiviral, Doxycycline inducible; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22552098
Proper citation: RRID:Addgene_38240 Copy
Species: Mus musculus
Genetic Insert: Smo
Vector Backbone Description: Vector Backbone:pGE; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:12414725
Proper citation: RRID:Addgene_38094 Copy
Species: Rattus norvegicus
Genetic Insert: microtubule-associated protein 1 light chain 3 beta
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:18443221
Proper citation: RRID:Addgene_38195 Copy
Species: Mus musculus
Genetic Insert: a novel protein kinase structurally related to C. elegans UNC-51
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:18443221
Proper citation: RRID:Addgene_38197 Copy
Species: Mus musculus
Genetic Insert: a novel protein kinase structurally related to C. elegans UNC-51
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:18443221
Proper citation: RRID:Addgene_38198 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: HIS3
Vector Backbone Description: Backbone Size:6614; Vector Backbone:pCENBEVY-U; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24357599
Proper citation: RRID:Addgene_38228 Copy
Species: Photinus pyralis
Genetic Insert: Firefly Luciferase
Vector Backbone Description: Backbone Size:7122; Vector Backbone:pCENBEVY-U; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24357599
Proper citation: RRID:Addgene_38223 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: Saccharomyces cerevisiae
Genetic Insert: GCN4-based single uORF-containing 5'-UTR
Vector Backbone Description: Backbone Size:6614; Vector Backbone:pCENBEVY-U; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24357599
Proper citation: RRID:Addgene_38220 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: Mus musculus
Genetic Insert: PTK2 protein tyrosine kinase 2
Vector Backbone Description: Backbone Marker:Dr. Toshio Kitamura of the University of Tokyo ; Backbone Size:6878; Vector Backbone:pMXs-puro; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:18443221
Proper citation: RRID:Addgene_38194 Copy
Species: Photinus pyralis
Genetic Insert: Firefly Luciferase
Vector Backbone Description: Backbone Size:6616; Vector Backbone:pCENBEVY-U; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24357599
Proper citation: RRID:Addgene_38219 Copy
Species: Photinus pyralis
Genetic Insert: Firefly Luciferase
Vector Backbone Description: Backbone Size:6616; Vector Backbone:pCENBEVY-U; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24357599
Proper citation: RRID:Addgene_38217 Copy
Species: Photinus pyralis
Genetic Insert: Firefly Luciferase
Vector Backbone Description: Backbone Size:6616; Vector Backbone:pCENBEVY-U; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24357599
Proper citation: RRID:Addgene_38213 Copy
Species: Homo sapiens
Genetic Insert: BCR GAP domain encoding residues 1004-1271 with R1090A point mutation
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.
pcDNA3.1Xpress-BcrGAP plasmid(AddGene #38175) was digested by BstEII x EcoRI and a 1.2 kb fragment containing the R1090 wt site, TAG and 3’ UT sequences removed. The wt fragment was replaced by a 0.6 kb BstEII x EcoRI fragment containing the R1090A mutation isolated from EGFP-BCR R1090A pEGFP (AddGene #38173). The construct contains the natural TAG and 3’ UT sequences. The difference between wt and mutant plasmids is in the 3’ UT region. The construct includes amino acid residues 1004-1271.
Proper citation: RRID:Addgene_38176 Copy
Species: Homo sapiens
Genetic Insert: ABR DH domain N282AE283A
Vector Backbone Description: Backbone Size:5000; Vector Backbone:pGEX 3X; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17116687
Comments: Constructed by Young Jin Cho.
Mutagenesis done using Stratagene Quick Change kit.
0.35 kb Bgl IIxSstI fragment containing the N282A/E283A double mutation from Abr EYFP (AddGene #36419) was ligated into the "ABR DH domain pGEX" plasmid (AddGene #38181) digested with Bgl II x SstI and removal of the 0.35 kb fragment containing the wt sequence N282E283. The amino acid numbering is as in NM_021962 (isoform A).
Proper citation: RRID:Addgene_38182 Copy
Genetic Insert: Bxb1-Excisionase
Vector Backbone Description: Backbone Size:2800; Vector Backbone:J64100; Vector Types:Synthetic Biology, BioBrick cloning vector; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:22615351
Comments: In the reset circuit, PBAD controls expression of a polycistron encoding excisionase with a strong RBS followed by Bxb1 integrase with a GTG start codon to decrease its translation rate
and increase the excisionase to integrase ratio. This construct was cloned in a plasmid
with a regulated ColE1 origin of replication (J64100, 50-70 copies, chloramphenicol resistance).
Proper citation: RRID:Addgene_38207 Copy
Genetic Insert: Bxb1 Integrase
Vector Backbone Description: Vector Backbone:pSB3K1; Vector Types:Synthetic Biology, BioBrick cloning vector; Bacterial Resistance:Kanamycin
Defining Citation: PMID:22615351
Comments: The Bxb1 integrase was cloned downstream of the PBAD/AraC promoter (BBa_I0500) on pSB3K1 plasmid bearing a p15A origin of replication (15-20 copies). This version of the integrase has a 6-His-tag which we found to stabilize the protein. Therefore, a weak RBS (BBa_B0031) and a LAA ssrA tag was added to reduce the basal expression of the enzyme.
Proper citation: RRID:Addgene_38209 Copy
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