MDH1-PGK-H2Kb2
(Plasmid #17852)

• Depositing Lab: Chang-Zheng Chen
• Publication: Li et al Cell. 2007 Apr 6. 129(1):147-61.

Backbone

• Vector backbone: MDH1-404
• Backbone manufacturer: Chen lab
• Backbone size w/o insert (bp): 7200
• Vector type: Mammalian Expression, Retroviral
• Selectable markers: Blasticidin

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: H-2Kb
• Species: M. musculus (mouse)
• Insert Size (bp): 1000
• Entrez Gene: H2-K1 (a.k.a. H-2K, H-2K(d), H2-D1, H2-K, K-f)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: See map (not destroyed)
• 3′ cloning site: See map (not destroyed)
• 5′ sequencing primer: H1

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

The original retroviral miRNA expression construct (Chen et al., 2004) was modified
to incorporate a truncated H-2Kb surface marker and a blasticidin drug resistance gene to allow for magnetic bead assisted sorting and drug selection. These
marker genes are driven by the PGK promoter and transcribed as a bicistronic
message, with the truncated H-2Kb gene placed after the PGK promoter and the
blasticidin resistance gene after the EMCV IRES.
MiRNA gene expression is driven by the human H1 pol III promoter. The truncated H-2Kb selection marker is non-functional since it is devoid of a cytoplasmic domain and is of a different MHC haplotype than the T cells in 5C.C7 transgenic mice (H-2Kk).