pCI-neo-RL-ZEB2 200bmut
(Plasmid #35538)

• Depositing Lab: Greg Goodall
• Publication: Gregory et al Nat Cell Biol. 2008 May;10(5):593-601. Epub 2008 Mar 30.

Backbone

• Vector backbone: pCI-neo-RL
• Backbone manufacturer: Witold Filipowicz
• Backbone size w/o insert (bp): 6380
• Total vector size (bp): 8144
• Modifications to backbone: RL was inserted into pcI-neo(Promega) to form the plasmid pcI-neo-Rl by Witold Filipowicz. Pillai R, Bhattacharyya S, Artus C, Zoller T, Cougot N, Basyuk E, Bertrand E, Filipowicz W (2005) Inhibition of translational initiation by let-7 microRNA in human cells. Science 309:1573-1576
• Vector type: Mammalian Expression, Luciferase
• Selectable markers: Neomycin (select with G418)

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: 3'UTR of ZEB2 gene
• Alt name: HRIHFB2411, HSPC082, SIP-1, SIP1, SMADIP1, ZFHX1B
• Species: H. sapiens (human)
• Insert Size (bp): 1264
• Mutation: 6 Mutations in the miR-200b seed regions of the ZEB2 3'UTR
• GenBank ID: 9839
• Entrez Gene: ZEB2 (a.k.a. HSPC082, SIP-1, SIP1, SMADIP1, ZFHX1B)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XbaI (destroyed during cloning)
• 3′ cloning site: ZbaI (destroyed during cloning)
• 5′ sequencing primer: unknown
• 3′ sequencing primer: unknown

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Renilla Luciferase Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Mutations in the miR-200b seed regions of the ZEB2 3'UTR were generated using the QuikChange Multi site-directed mutagenesis kit.

The ZEB2 3'UTR was cloned into the plasmid pCIneo-RL which was obtained from Witold Filiowicz.
Pillai R, Bhattacharyya S, Artus C, Zoller T, Cougot N, Basyuk E, Bertrand E, Filipowicz W (2005) Inhibition of translational initiation by let-7 microRNA in human cells.
Science 309:1573-1576

Please note that Addgene's sequencing results identified a single nucleotide deletion at bp# 2440 when compared to the full plasmid sequence provided by the depositing laboratory. Addgene's sequencing results are correct as the extra base in the sequence from the depositing laboratory was an error due to a sequencing artifact.