pJH200
(Plasmid #13327)

• Depositing Lab: Martin Gellert
• Publication: Lieber et al Genes Dev. 1987 Oct . 1(8):751-61.

Backbone

• Vector backbone: n/a

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: recombination substrate

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: n/a

Terms and Licenses

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

Depositor Comments

pJH200 is identical to pJH201, except that pJH200 does not include the Ig kappa enhancer.

pJH201's construction was described in Hesse et al. Cell, 1987. Briefly, the early region of polyoma virus (bases 4632-2962) was isolated by BamHI-HincII digestion of PY-pBR322. The cat structural gene (bases 4241-5027) was isolated by DpnI-HindIII digestion of pRSVcat (Gorman et al., 1982). The mouse immunoglobulin kappa gene enhancer was isolated by AluI digestion of pHBC kappa (Lewis et al., 1982). The immunoglobulin joining signals in inverted orientation relative to each other and with heptamers fused were isolated by HindIII-ApaI digestion of the f-T subclone (Hochtl et al., 1982). A synthetic oligonucleotide was used to insert the oop transcription terminator of bacteriophage lambda, present on a 189 bp DpnI fragment (bases 38,478 - 38,666), between the joining signals, which were separated by cleavage with HgiAI at the junction of the fused heptamers. The resulting fragment and other component fragments were assembled in stepwise fashion using the multipurpose cloning site of pUC13 to yield pJH201.