TET-pLKO.1 PURO shSMAD4 #1
(Plasmid #83089)

• Purpose: Lentiviral shRNA vector for inducible knockdown of human SMAD4 (cross reacts with mouse Smad4)
• Depositing Lab: Kevin Janes
• Publication: Bajikar et al Dev Cell. 2017 Nov 20;43(4):418-435.e13. doi: 10.1016/j.devcel.2017.10.027.

Backbone

• Vector backbone: Tet-pLKO-puro
• Backbone manufacturer: Addgene #21915
• Backbone size w/o insert (bp): 10633
• Total vector size (bp): 8816
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: shSMAD4
• gRNA/shRNA sequence: GCCAGCTACTTACCATCATAA
• Species: H. sapiens (human), M. musculus (mouse)
• GenBank ID: NM_005359 NM_008540
• Entrez Gene: SMAD4 (a.k.a. DPC4, JIP, MADH4, MYHRS)
• Entrez Gene: Smad4 (a.k.a. D18Wsu70e, DPC4, Madh4)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: AgeI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: GGCAGGGATATTCACCATTATCGTTTCAGA

Terms and Licenses

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

Depositor Comments

The targeting sequence was identified from the TRC (TRCN0000025900). Potently knocks down SMAD4 expression after 3 x 500 uL lentiviral infection in MDA-MB-231 cells.