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PurposeBackbone pComb3X containing PCR template for human antibody lambda light chain constant region
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 63892 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepComb3X
- Backbone size w/o insert (bp) 3300
- Total vector size (bp) 4700
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Vector typeBacterial Expression ; phage display
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)XL1 Blue
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert namehuman antibody lambda light chain
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SpeciesH. sapiens (human)
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Insert Size (bp)320
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Entrez GeneIGLL5 (a.k.a. IGL, IGLV, VL-MAR)
- Promoter lacZ
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site SfiI (not destroyed)
- 3′ cloning site SfiI (not destroyed)
- 5′ sequencing primer AAG ACA GCT ATC GCG ATT GCA G
- 3′ sequencing primer GCC CCC TTA TTA GCG TTT GCC ATC (Common Sequencing Primers)
Resource Information
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Article Citing this Plasmid
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor 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. pComb3XLambda can be used as a template to amplify the lambda light chain constant region for construction of chimeric antibody libraries as described in:
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
Please note that there is a A246V mutation in heavy chain region; however, this vector is meant to be used as a PCR template for the light chain region from base# 613-1016.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
pComb3XLambda was a gift from Carlos Barbas (Addgene plasmid # 63892 ; http://n2t.net/addgene:63892 ; RRID:Addgene_63892) -
For your References section:
Methods for the generation of chicken monoclonal antibody fragments by phage display. Andris-Widhopf J, Rader C, Steinberger P, Fuller R, Barbas CF 3rd. J Immunol Methods. 2000 Aug 28;242(1-2):159-81. 10.1016/s0022-1759(00)00221-0 PubMed 10986398