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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: 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
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
Vector Backbone Description: Backbone Size:6700; Vector Backbone:pcDNA3.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17116687
Comments: Constructed by Young Jin Cho.
A 1.2 kb BamHI xEcoRI fragment from BcrGAP in pGEX3x (AddGene #36405) was inserted into pcDNA BamHI xEcoRI. Contains natural TAG and 3’ UT sequences. The construct includes amino acid residues 1004-1271.
Proper citation: RRID:Addgene_38175 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
Species: Synthetic
Genetic Insert: mKate2
Vector Backbone Description: Vector Backbone:Phi80; Vector Types:Synthetic Biology, Integration vector; Bacterial Resistance:Kanamycin
Defining Citation: PMID:22615351
Comments: This strain contains Addgene plasmid 38200 chromosomally integrated at the Phi80 (Haldimann et al., J of Bacteriology 2001) site of DH5alphaZ1 strain.
Proper citation: RRID:Addgene_38203 Copy
Species: Mus musculus
Genetic Insert: Unc-51 like kinase 2
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_38206 Copy
Species: Homo sapiens
Genetic Insert: FER
Vector Backbone Description: Backbone Marker:Pharmingen; Backbone Size:8530; Vector Backbone:pAcG2T; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Comments: Constructed by Leena Haataja. 4-way ligation of BamHI/NsiI + NsiI/KpnI + KpnI/EcoRI fragments into pAcG2T digested with BamHI x EcoRI. The construct makes Fer protein but not from amplified virus stocks. Large-scale yield of protein is relatively low.
Proper citation: RRID:Addgene_38167 Copy
Species: Synthetic
Genetic Insert: mKate2
Vector Backbone Description: Vector Backbone:Phi80; Vector Types:Synthetic Biology, Integration vector; Bacterial Resistance:Kanamycin
Defining Citation: PMID:22615351
Comments: This construct has attL and attR sites (LR).
To integrate on your favorite E.coli strain, transform in strain containing the pAH123 plasmid (Haldimann et al., J of Bacteriology 2001). Contains a R6K origin of replication, will only replicate in pir2+ strains.
Proper citation: RRID:Addgene_38200 Copy
Species: Mus musculus
Genetic Insert: Unc-51 like kinase 2
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_38201 Copy
Species: Synthetic
Genetic Insert: mKate2
Vector Backbone Description: Vector Backbone:Phi80; Vector Types:Synthetic Biology, Integration vector; Bacterial Resistance:Kanamycin
Defining Citation: PMID:22615351
Comments: This strain contains Addgene plasmid 38170 chromosomally integrated at the Phi80 (Haldimann et al., J of Bacteriology 2001) site of DH5alphaZ1 strain.
Proper citation: RRID:Addgene_38202 Copy
Species: Homo sapiens
Genetic Insert: ABR
Vector Backbone Description: Backbone Marker:Pharmingen; Backbone Size:8530; Vector Backbone:pAcG2T; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Comments: Constructed by Leena Haataja. 4-way ligation of BamHI/StuI + StuI/BstEII + BstEII/EcoRI fragments into pAcG2T digested with BamHI x EcoRI. The construct makes Abr protein.
Addgene sequencing found an E24G point mutation--this does not alter plasmid function.
Proper citation: RRID:Addgene_38162 Copy
Species: Homo sapiens
Genetic Insert: BCR
Vector Backbone Description: Backbone Marker:Pharmingen; Backbone Size:8530; Vector Backbone:pAcG2T; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Comments: Constructed by Vesa Kaartinen. Makes Bcr protein. Nucleotide sequence from BamHI cloning site to BCR ATG is:
5' GGA TCC CCC GGG CTG CAG GAA TTC GAT GGC CGC GCC ATG
Proper citation: RRID:Addgene_38163 Copy
Species: Homo sapiens
Genetic Insert: Rac1
Vector Backbone Description: Backbone Marker:Pharmingen; Backbone Size:8530; Vector Backbone:pAcG2T; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Comments: Constructed by Nora Heisterkamp. Rac1 cDNA isolated from K562 cDNA library. 5' BamHI site introduced by PCR. Constructed in 3-way ligation: 0.18 kb 5' BamHI-PvuII + 3' 0.4 kb PvuII-EcoRI into pAcG2T. Makes Rac1 in insect cells.
Proper citation: RRID:Addgene_38164 Copy
Species: Homo sapiens
Genetic Insert: ARRDC3
Vector Backbone Description: Backbone Marker:Invitrogen; Vector Backbone:pEF6; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20559325
Proper citation: RRID:Addgene_38317 Copy
Species: Homo sapiens
Genetic Insert: TSG101
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5780; Vector Backbone:pEF6; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22315426
Comments: Please note that there is a mutation in the TSG101 ORF, Y82H. This mutation should not affect plasmid function.
Proper citation: RRID:Addgene_38318 Copy
Species: Homo sapiens
Genetic Insert: ARRDC1
Vector Backbone Description: Backbone Marker:Promega; Backbone Size:5472; Vector Backbone:pCI-neo; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22315426
Proper citation: RRID:Addgene_38319 Copy
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