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Vector Backbone Description: Vector Backbone:pST50Trc; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15766881
Comments: Replace DHFR with GOI using BamHI/NgoMIV; Clone out cassette using XbaI/BamHI
Proper citation: RRID:Addgene_63935 Copy
Vector Backbone Description: Vector Backbone:pST50Trc; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15766881
Comments: Replace DHFR with GOI using BamHI/NgoMIV; Clone out cassette using XbaI/BamHI
Proper citation: RRID:Addgene_63937 Copy
Vector Backbone Description: Vector Backbone:pST50Trc; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15766881
Comments: Replace DHFR with GOI using BamHI/NgoMIV; Clone out cassette using XbaI/BamHI
Proper citation: RRID:Addgene_63938 Copy
Species: Mus musculus
Genetic Insert: Nanog homeobox
Vector Backbone Description: Backbone Marker:Promega; Backbone Size:5256; Vector Backbone:pGL3-Control; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22020437
Comments: Addgene's quality control sequence with EBV-rev primer shows a single bp mismatch with the available reference sequence.
Proper citation: RRID:Addgene_63893 Copy
Species: Mus musculus
Genetic Insert: Nanog homeobox
Vector Backbone Description: Backbone Marker:Promega; Backbone Size:5256; Vector Backbone:pGL3-Control; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22020437
Comments: Mir-34a binding site was mutated to remove the seed region
Proper citation: RRID:Addgene_63894 Copy
Species: Synthetic
Genetic Insert: SS Stuffer
Vector Backbone Description: Backbone Size:3300; Vector Backbone:pComb3H; Vector Types:Bacterial Expression, phage display; Bacterial Resistance:Ampicillin
Defining Citation: PMID:10986398
Comments: pComb3X is the newest of the pComb vectors. Improvements over pComb3 include increased stability and introduction of an asymmetric SfiI cassette for directional cloning of full Fab, scFv, peptide and other protein for phage display. 6xHis and HA tags allow for purification and detection. An amber stop codon was introduced to turn-off expression of the pIII fusion protein by switching to a non-supressor strain of E. coli allowing production of soluble protein without subcloning. Alternatively, the gene for phage protein pIII can be removed by SpeI/NheI digest. pComb3XSS is recommended for preparation of vector for library cloning. The “SS” refers to the double stuffer, a 1200bp stuffer in the Fab light chain cloning region bounded by SacI and XbaI restriction sites and a 300bp stuffer in Fab heavy chain cloning region bound by XhoI and SpeI restriction sites. Also, the 1600bp double stuffer (both stuffers plus the leader sequence between the Fab light chain and heavy chain cloning regions) can be removed by SfiI digest so that non-Fab genes of interest can be cloned. Also available on Addgene: pComb3XTT and pComb3XLambda are only needed at templates for the construction of chimeric Fab libraries as described in Phage Display: A Laboratory Manual. pComb3XTT can also be used as an Fab expression control.
Select References:
Barbas, C. F., III; Burton, D. R.; Scott, J.K., Silverman, G.J. Eds. (2001) Phage Display: A Laboratory Manual; Cold Spring Harbor Laboratory Press: Cold Spring Harbor, New York
Rader, C; Popkov, M.; Neves, J.A.; Barbas III, C.F. (2002) Integrin _v_3 Targeted Therapy of Kaposi’s Sarcoma with an In Vitro Evolved Antibody. FASEB, 16(14):2000-2.
Berry, J.D.; Rutherford, J.; Silverman, G.J.; Kaul, R.; Elia, M.; Gobuty, S.; Fuller, R.; Plummer, F.A.; & Barbas III, C.F. (2003) Development of Functional Human Monoclonal Single-Chain Variable Fragment Antibody Against HIV-1 From Human Cervical B cells. Hybridoma and Hybridomics, 22(2):, 97-108.
Jendreyko N, Popkov M, Beerli RR, Chung J, McGavern DB, Rader C, Barbas CF 3rd. (2003) Intra-diabodies: Bispecific, tetravalent antibodies for the simultaneous functional knockout of two cell surface receptors. J Biol Chem., 278(48):, 47812-9.
Steinberger, P.; Sutton, J.K.; Rader, C.; Elia, M.; and Barbas III, C. F. (2000) Generation and Characterization of a Recombinant Human CCR5-specific Antibody: A Phage Display Approach for Rabbit Antibody Humanization. J. Biol. Chem., 275,, 36073-36078.
Goncalves, J.; Kilva, F.; Freitas-Vieira, A.; Santa-Marta, M.; Malho, R.; Yang, X.; Gabuzda, D.; and Barbas III, C.F. (2002) Functional Neutralization of HIV-1 Vif Protein by Intracellular Immunization Inhibits Reverse Transcription and Viral Replication. J. Biol. Chem., 277(35):32036-45.
Popkov, M.; Mage, R.G.; Alexander, C.B.; Thundivalappil, S.; Barbas III, C.F.; Rader, C. (2003) Rabbit immune repertoires as sources for therapeutic monoclonal antibodies: The impact of Kappa Allotype-correlated variation in cysteine content on antibody libraries selected by phage display. J. Mol. Biol., 325:325-335.
Chung, J.; Rader, C.; Popkov, M.; Hur, Y.-M.; Kim, H.-K.; Lee, Y.-J.; & Barbas III, C. F. (2004) Integrin _IIb_3 specific synthetic human monoclonal antibodies and HCDR3 peptides that potently inhibit platelet aggregation,FASEB 18(2):361-3.
Popkov, M.; Jendreyko, N.; Gonzalez-Sapienza, G.; Mage, R.G.; Rader, C.; Barbas III, C.F. (2004) Human/mouse cross-reactive anti-VEGF receptor 2 recombinant antibodies selected from animmune b9 allotype rabbit antibody library, J. Immunol. Methods, 288(1-2):149-164.
Popkov, M.; Rader, C.; Barbas III, C.F. (2004) Isolation of human prostate cancer reactive
antibodies using phage display technology, J. Immunol. Methods, 291:137-151.
Tanaka, F.; Fuller, R.; Barbas III, C.F. (2005) Development of Small Designer Aldolase Enzymes: Catalytic Activity, Folding, and Substrate Specificity. Biochemistry, 44:7583-7592.
This plasmid has been found to be prone to multimerization. Multimerization often does not impact plasmid function, but may reduce transformation efficiencies. See our article on Plasmid Dimers and Multimers for more information (https://blog.addgene.org/plasmids-101-dimers-and-multimers)
Proper citation: RRID:Addgene_63890 Copy
Vector Backbone Description: Vector Backbone:pST50Trc; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15766881
Comments: Replace DHFR with GOI using BamHI/NgoMIV; Clone out cassette using XbaI/BamHI
Proper citation: RRID:Addgene_63944 Copy
Vector Backbone Description: Vector Backbone:pST50Trc; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15766881
Comments: Replace DHFR with GOI using BamHI/NgoMIV; Clone out cassette using XbaI/BamHI
Proper citation: RRID:Addgene_63943 Copy
Vector Backbone Description: Vector Backbone:pST50Trc; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15766881
Comments: Replace DHFR with GOI using BamHI/NgoMIV; Clone out cassette using XbaI/BamHI
Proper citation: RRID:Addgene_63948 Copy
Vector Backbone Description: Vector Backbone:pST50Trc; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15766881
Comments: Replace DHFR with GOI using BamHI/NgoMIV; Clone out cassette using XbaI/BamHI
Proper citation: RRID:Addgene_63949 Copy
Vector Backbone Description: Vector Backbone:pST50Trc; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15766881
Comments: Replace DHFR with GOI using BamHI/NgoMIV; Clone out cassette using EcoRI/HindIII
Proper citation: RRID:Addgene_63955 Copy
Vector Backbone Description: Vector Backbone:pST50Trc; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15766881
Comments: Replace DHFR with GOI using BamHI/NgoMIV; Clone out cassette using EcoRI/HindIII
Proper citation: RRID:Addgene_63958 Copy
Vector Backbone Description: Vector Backbone:pST50Trc; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15766881
Comments: Replace DHFR with GOI using BamHI/NgoMIV; Clone out cassette using XbaI/BamHI
Proper citation: RRID:Addgene_63952 Copy
Vector Backbone Description: Vector Backbone:pST50Trc; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15766881
Comments: Replace DHFR with GOI using BamHI/NgoMIV; Clone out cassette using XbaI/BamHI
Proper citation: RRID:Addgene_63950 Copy
Vector Backbone Description: Vector Backbone:pST50Trc; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15766881
Comments: Replace DHFR with GOI using BamHI/NgoMIV; Clone out cassette using EcoRI/HindIII
Proper citation: RRID:Addgene_63969 Copy
Vector Backbone Description: Vector Backbone:pST50Trc; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15766881
Comments: Replace DHFR with GOI using BamHI/NgoMIV; Clone out cassette using EcoRI/HindIII
Proper citation: RRID:Addgene_63965 Copy
Vector Backbone Description: Vector Backbone:pST50Trc; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15766881
Comments: Replace DHFR with GOI using BamHI/NgoMIV; Clone out cassette using SacI/KpnI
Proper citation: RRID:Addgene_63975 Copy
Vector Backbone Description: Vector Backbone:pST50Trc; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15766881
Comments: Replace DHFR with GOI using BamHI/NgoMIV; Clone out cassette using EcoRI/HindIII
Proper citation: RRID:Addgene_63970 Copy
Species: Homo sapiens
Genetic Insert: TT Fab
Vector Backbone Description: Backbone Size:3500; Vector Backbone:pAra; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Comments: Provided with the pComb3H system for arabinose induced expression of soluble protein with detection tags. The “TT” refers to the human Fab to tetanus toxin which can be used as a protein expression control. Alternatively, one could upgrade to the pComb3X system, also available on Addgene, which already contains tags.
Select reference:
Cary, S.P.; Lee, J.; Wagenknecht, R.; Silverman, G.J. (2000) Characterization of Superantigen-Induced Clonal Deletion with a Novel Clan III-Restricted Avian Monoclonal Antibody: Exploiting Evolutionary Distance to Create Antibodies Specific for a Conserved VH Region Surface. J. Immun., 164, 4730-4741.
Proper citation: RRID:Addgene_63901 Copy
Species: Escherichia coli
Genetic Insert: lacZ
Vector Backbone Description: Backbone Size:4500; Vector Backbone:pACYC184; Vector Types:Bacterial Expression; Bacterial Resistance:Tetracycline
Defining Citation: PMID:9092628
Comments: AUG --> GUG start codon in lacZ
Proper citation: RRID:Addgene_63866 Copy
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