pLOX-TERT-iresTK
(Plasmid #12245)

• Purpose: 2nd generation lentiviral vector for reversible immortalization of human primary cells
• Depositing Lab: Didier Trono
• Publication: Salmon et al Mol Ther. 2000 Oct . 2(4):404-14.

Backbone

• Vector backbone: pHR'
• Backbone size w/o insert (bp): 13115
• Vector type: Mammalian Expression, Lentiviral, Cre/Lox

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Growth instructions: Use Stbl3 or HB101 to reduce chance of recombination. Grow at 37C
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: TERT, HSV1-TK
• Alt name: TERT
• Alt name: HSV-1 thymidine kinase
• Species: H. sapiens (human)
• Mutation: LoxP in 3' LTR.
• Entrez Gene: TERT (a.k.a. CMM9, DKCA2, DKCB4, EST2, PFBMFT1, TCS1, TP2, TRT, hEST2, hTRT)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: CMV Fwd for hTert
• 3′ sequencing primer: SP6 primer rev for LoxP

Resource Information

• Articles Citing this Plasmid:
  • 27 References

Terms and Licenses

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

Depositor Comments

CMV promoter is upstream of the bicistronic coding cassette with hTert and HSV1-TK. During reverse transcription U3 region of the 3'LTR, with LoxP, is duplicated; LoxP sites end up flanking the genome of the integrated provirus. Upon expression of Cre, provirus is excised.

Please visit the Trono lab http://tronolab.epfl.ch for cloning strategies, protocols, publications, and more.