pCL-CTIG
(Plasmid #14901)

• Purpose: 3rd generation lentiviral vector expressing tTA-VP16 and EGFP.
• Depositing Lab: Inder Verma
• Publication: Kafri et al Mol Ther. 2000 Jun . 1(6):516-21.

Backbone

• Vector backbone: pCL-CG
• Backbone manufacturer: Verma Lab
• Backbone size w/o insert (bp): 8600
• Vector type: Mammalian Expression, Lentiviral

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: CMV-tTA-GFP
• Alt name: CMV GFP
• Alt name: GFP
• Insert Size (bp): 3000

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (destroyed during cloning)
• 3′ cloning site: Eco47 (destroyed during cloning)
• 5′ sequencing primer: n/a

Resource Information

• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

This tet-inducible vector was constructed by cloning an XbaI/HpaI fragment containing the tTA and the
inducible promoter from pBStind into NheI/Eco47(AfeI) sites of pCL-CG. The inducible cassette contains the tet dependent
transactivator (tTA) at its 5' end and an
inducible promoter at its 3' end. Expression
of the GFP gene is controlled by the inducible promoter,
which contains a minimal promoter (mp) and seven
copies of Tet operator (tetO). The tTA gene is constitutively
expressed under the control of the CMV promoter.
Following transduction, the constitutively
expressed tTA (tetRVP16) binds to tet(O)- mp and
induces the transcription of a high level of GFP mRNA.
In contrast, binding of doxycycline to the tTA induces
conformational changes that prevent it from binding to
the tet(O)- minimal promoter and consequently leads to
little or no GFP expression.