pJH298
(Plasmid #115651)

• Purpose: recombination substrate
• Depositing Lab: Martin Gellert
• Publication: Lieber et al Cell. 1988 Oct 7. 55(1):7-16.

Backbone

• Vector backbone: pUC13

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: V(D)J recombination substrate

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: unknown (unknown if destroyed)
• 3′ cloning site: unknown (unknown if destroyed)
• 5′ sequencing primer: unknown

Terms and Licenses

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

Depositor Comments

The plasmid backbone is a fusion of pUC13 and the early region of the polyoma virus genome, which allows the plasmids to replicate autonomously in E. coli and murine cells, respectively. Various DNA fragments pertinent to this recombination assay are inserted into the multipurpose cloning site of pUC13, including a prokaryotic promoter, the structural gene encoding chloramphenicol acetyltransferase (cat), and the intervening transcription terminator. The terminator is flanked on the promoter-proximal side by a unique SalI site and on the cat gene side by a unique BamHI site. DNA fragments containing V(D)J recombination signals are inserted into these unique sites. (These signals are the only DNA segments related to Ig genes or TCR genes included on the plasmid). A 39-bp fragment containing a 12-signal, created by annealing of synthesized complementary oligonucleotides with protruding ends compatible with SalI, is inserted into the SalI site of the backbone. A 49bp fragment containing a 23-signal, made similarly but with protruding ends compatible with BamHI, is inserted into the BamHI site of the backbone.
 
pJH298 contains signals bearing the consensus versions of heptamers and nonamers.