pComb3XLambda
(Plasmid #63892)

• Purpose: Backbone pComb3X containing PCR template for human antibody lambda light chain constant region
• Depositing Lab: Carlos Barbas
• Publication: Andris-Widhopf et al J Immunol Methods. 2000 Aug 28;242(1-2):159-81.

Backbone

• Vector backbone: pComb3X
• Backbone size w/o insert (bp): 3300
• Total vector size (bp): 4700
• Vector type: Bacterial Expression ; phage display

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL1 Blue
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: human antibody lambda light chain
• Species: H. sapiens (human)
• Insert Size (bp): 320
• Entrez Gene: IGLL5 (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

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

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.