pCMVhTRIM65_3Flag
(Plasmid #53966)

• Purpose: Human TRIM65 mammalian expression vector with 3xFLAG epitope at c-terminus.
• Depositing Lab: Martin Dorf
• Publication: Li et al Proc Natl Acad Sci U S A. 2014 May 13;111(19):6970-5. doi: 10.1073/pnas.1322545111. Epub 2014 Apr 28.

Backbone

• Vector backbone: pCMV-3Tag-8
• Backbone manufacturer: Agilent (formerly Stratagene)
• Backbone size w/o insert (bp): 5171
• Total vector size (bp): 6725
• Modifications to backbone: Contruct: [XhoI site]CTCGAG-CTG-[HindIII]AAGCTT-(Kozak seq]GCC-insert[hTRIM65 CDS without stop, 1551bp]-[Clai]ATCGAT. The commercial multiple cloning site has been replaced as indicated. Backbone's 3xFLAGs are in frame with the restriction sites.
• Vector type: Mammalian Expression
• Selectable markers: Hygromycin

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: TRIM65
• Alt name: tripartite motif containing 65
• Species: H. sapiens (human)
• Insert Size (bp): 1554
• GenBank ID: BC021259.2 has same CDS.
• Entrez Gene: TRIM65 (a.k.a. 4732463G12Rik)
• Promoter: CMV
• Tag / Fusion Protein:
  • 3x FLAG (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: Clai (not destroyed)
• 5′ sequencing primer: CMV, T3 (28bp from insert)
• 3′ sequencing primer: T7, BGH reverse

Terms and Licenses

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

Depositor Comments

Cloned from human T98 glioblastoma line. The coding sequence is identical to that in BC021259.2