pBudORF1-CH
(Plasmid #51290)

• Purpose: expresses the codon optimized ORF1 of L1RP with his tag
• Depositing Lab: Astrid Roy-Engel
• Publication: Wagstaff et al Mol Biol Evol. 2013 Jan;30(1):88-99. doi: 10.1093/molbev/mss202. Epub 2012 Aug 23.

Backbone

• Vector backbone: pBudCE4.1
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 4595
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Bleocin (Zeocin), 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: L1 ORF1
• Alt name: L1RE1
• Alt name: Line1
• Species: H. sapiens (human)
• Entrez Gene: L1RE1 (a.k.a. L1.2, LRE1)
• Promoter: CMV
• Tag / Fusion Protein:
  • HIS (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: EBV-rev

Terms and Licenses

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

Depositor Comments

Although a myc tag is present in the full sequence, this tag is not present in this plasmid.

pBudORF1-CH was generated by cloning the polymerase chain reaction (PCR)-amplified codon-optimized consensus sequences of each ORF into the HindIII-BamHI sites of the pBudCE4.1 vector in a manner that removes the stop codon of the ORF1, so the expressed protein will contain the his tag at the carboxy terminus. The following primers were used in the amplification of ORF1: 5-AGACCCAAGCTTAGCTAAAACCACAAAGATG-3′ and 5-TGTTCGGATCCGATCTTGGTGTGCTTCTGCAGGGG-3′