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Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:2720; Vector Backbone:pENTR4; Vector Types:Mammalian Expression, RNAi; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21418658
Proper citation: RRID:Addgene_32686 Copy
Genetic Insert: ECFP
Vector Backbone Description: Backbone Marker:Invitrogen; Vector Backbone:pDONR221 P4r-P3r; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21772812
Proper citation: RRID:Addgene_32594 Copy
Genetic Insert: IRES-EGFP
Vector Backbone Description: Backbone Marker:Invitrogen; Vector Backbone:pDONR221 P3-P2; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21772812
Comments: BMC Biotechnol. 2006 Jan 12;6:4.
Development of a new bicistronic retroviral vector with strong IRES activity.
Martin P, Albagli O, Poggi MC, Boulukos KE, Pognonec P.
Source
CNRS UMR6548, Parc Valrose, Université de Nice Sophia Antipolis, Nice, France. [email protected]
Proper citation: RRID:Addgene_32591 Copy
Species: Homo sapiens
Genetic Insert: Synapsin I promoter
Vector Backbone Description: Backbone Marker:Invitrogen; Vector Backbone:pDONR221 P1-P5r; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21772812
Comments: Gene Therapy 14, 872-882 (June 2007) doi:10.1038/sj.gt.3302924
Efficient gene transduction of neurons by lentivirus with enhanced neuron-specific promoters
H Hioki, H Kameda, H Nakamura, T Okunomiya, K Ohira, K Nakamura, M Kuroda, T Furuta and T Kaneko
Proper citation: RRID:Addgene_32582 Copy
Genetic Insert: Intronic Luciferase miRNA and EGFP
Vector Backbone Description: Backbone Marker:Invitrogen; Vector Backbone:pDONR221 P3-P2; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21772812
Comments: LucmiRNA atttgtattcagcccatatcgg
Du, G., Yonekubo, J., Zeng, Y., Osisami, M., and Frohman, M. A. (2006). Design of expression vectors for RNA interference based on miRNAs and RNA splicing. FEBS J. 273, 5421–5427.
Proper citation: RRID:Addgene_32588 Copy
Genetic Insert: Intronic HCN1miRNA and EGFP
Vector Backbone Description: Backbone Marker:Invitrogen; Vector Backbone:pDONR221 P4r-P3r; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21772812
Comments: HCN1miRNA TAAAGACGACAGTAGGTATCAG
Du, G., Yonekubo, J., Zeng, Y., Osisami, M., and Frohman, M. A. (2006). Design of expression vectors for RNA interference based on miRNAs and RNA splicing. FEBS J. 273, 5421–5427.
Proper citation: RRID:Addgene_32586 Copy
Species: Homo sapiens
Genetic Insert: MLH1
Vector Backbone Description: Backbone Marker:Structural Genomics Consortium (Addgene plasmid 26096); Backbone Size:7325; Vector Backbone:pET28-MHL; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Comments: PDB: 3RBN. http://www.thesgc.org/structures/3RBN/
Proper citation: RRID:Addgene_32859 Copy
Species: Homo sapiens
Genetic Insert: PANK1
Vector Backbone Description: Backbone Marker:Structural Genomics Consortium (Addgene plasmid 26094); Vector Backbone:pET28a-LIC; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Comments: PDB: 3SMP. http://www.thesgc.org/structures/3SMP/
Proper citation: RRID:Addgene_32871 Copy
Species: Homo sapiens
Genetic Insert: HECTD1
Vector Backbone Description: Backbone Marker:Structural Genomics Consortium (Addgene plasmid 26096); Backbone Size:7325; Vector Backbone:pET28-MHL; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Comments: PDB: 2LC3. http://www.thesgc.org/structures/2LC3/
Proper citation: RRID:Addgene_32860 Copy
Species: Toxoplasma gondii
Genetic Insert: MAPK2
Vector Backbone Description: Backbone Marker:Structural Genomics Consortium (Addgene plasmid 26096; Backbone Size:7325; Vector Backbone:pET28-MHL; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Comments: PDB: 3RP9. http://www.thesgc.org/structures/3RP9/
Proper citation: RRID:Addgene_32862 Copy
Species: Homo sapiens
Genetic Insert: SETD1A
Vector Backbone Description: Backbone Marker:Structural Genomics Consortium (Addgene plasmid 26096); Backbone Size:7325; Vector Backbone:pET28-MHL; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Comments: PDB: 3S8S. http://www.thesgc.org/structures/3S8S/
Proper citation: RRID:Addgene_32868 Copy
Species: Homo sapiens
Genetic Insert: TPK1
Vector Backbone Description: Backbone Marker:Structural Genomics Consortium (Addgene plasmid 26096); Backbone Size:7325; Vector Backbone:pET28-MHL; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Comments: PDB: 3S4Y. http://www.thesgc.org/structures/3S4Y/
Proper citation: RRID:Addgene_32865 Copy
Genetic Insert: GFP
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:3970; Vector Backbone:pmKate2-C; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21558267
Comments: The gene for dark GFP (S65T) (referred to as GFP from this point forward) was created by PCR amplifying the GFP gene using a reverse primer that also encoded the 27 amino acid quenching peptide derived from the transmembrane region of influenza M2. A sequence encoding the linker (LEVLFQGP) was then inserted between the EGFP and M2 genes using the QuikChange mutagenesis (Stratagene) approach. The newly inserted linker sequence was then mutated to encode the caspase-7 cleavage recognition site (DEVDFQGP). The final sequence of CA-GFP protein is GFP with the fusion of DEVDFQGPCNDSSDPLVVAASIIGILHLILWILDRL at the C terminus. This construct was used for expression and purification of CA-GFP.
Proper citation: RRID:Addgene_32748 Copy
Species: Homo sapiens
Genetic Insert: TDRD5
Vector Backbone Description: Backbone Marker:Structural Genomics Consortium (Addgene plasmid 26096); Backbone Size:7325; Vector Backbone:pET28-MHL; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Comments: PDB: 3S93. http://www.thesgc.org/structures/3S93/
Proper citation: RRID:Addgene_32869 Copy
Species: Mus musculus
Genetic Insert: Venus CAM KII Alpha
Vector Backbone Description: Backbone Size:3984; Vector Backbone:pEGFP C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:19339497
Proper citation: RRID:Addgene_29429 Copy
Species: Mus musculus
Genetic Insert: Venus CamKII alpha
Vector Backbone Description: Backbone Size:3984; Vector Backbone:pEGFP N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:19339497
Proper citation: RRID:Addgene_29427 Copy
Genetic Insert: Venus-17-Venus
Vector Backbone Description: Backbone Size:3984; Vector Backbone:pEGFP C1; Vector Types:Mammalian Expression, Anisotropy and brightness standard; Bacterial Resistance:Kanamycin
Defining Citation: PMID:19339497
Comments: Anisotropy standard and brightness standard
Linker sequence: TCCGGACTCAGATCTCGAGCTCAAGCTTCGAATTCTGCAGTCGACGGTACCAGCAAGG
Proper citation: RRID:Addgene_29424 Copy
Species: Homo sapiens
Genetic Insert: Metalloproteinase protein 8
Vector Backbone Description: Backbone Marker:Stratagene; Backbone Size:4300; Vector Backbone:pCMV-Tag4A; Vector Types:Mammalian Expression, Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:19330028
Proper citation: RRID:Addgene_29546 Copy
Species: Mus musculus
Genetic Insert: Kif2A-alpha
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4706; Vector Backbone:PEGFP-C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:15883193
Comments: Depositor believes the mutation in Kif2a is the results of either errors in database sequences or polymorphisms.
Proper citation: RRID:Addgene_29481 Copy
Genetic Insert: None
Vector Backbone Description: Backbone Size:8106; Vector Backbone:pET; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Comments: This plasmid is an empty vector. Your gene can be inserted with a SLIC cloning protocol.
Prescission is a highly specific protease that cleaves LEVLFQ/GP. It is generally more active than TEV protease, and many users have found that when their protein inhibits TEV cleavage, switching to a prescission vector has proved worthwhile.
To clone into this vector, add SLIC tags to the 5' end of your PCR primers.
Forward - 5'GTGCTGTTCCAGGGTCCGAAT3'
Reverse - 5'TGGTGGTGGTGGTGCTCGA(TTA)3'
Linearize the plasmid with SspI and XhoI, then gel purify. There is a 1650 bp stuffer sequence that will be visible on a gel.
When digesting the DNA with T4 polymerase, use no nucleotides for either your insert or the linearized vector.
More information on this vector can be found through http://qb3.berkeley.edu/qb3/macrolab/
Proper citation: RRID:Addgene_29721 Copy
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