pAd-M3cherry
(Plasmid #61041)

• Purpose: This plasmid mediates polycistronic expression of 4 genes: Ngn3, Pdx1, Mafa, and mCherry. It can be used to convert mouse pancreatic acinar cells to induced beta-like cells in vivo.
• Depositing Lab: Qiao Zhou
• Publication: Li et al Nat Biotechnol. 2014 Dec;32(12):1223-30. doi: 10.1038/nbt.3082. Epub 2014 Nov 17.

Backbone

• Vector backbone: pAd CMV/V5-DEST
• Backbone manufacturer: lifetechnologies
• Backbone size w/o insert (bp): 36686
• Total vector size (bp): 40200
• Modifications to backbone: none
• Vector type: Adenoviral

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: Ngn3
• Alt name: Neurog3
• Species: M. musculus (mouse)
• Insert Size (bp): 642
• Entrez Gene: Neurog3 (a.k.a. Atoh5, Math4B, bHLHa7, ngn3)

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: salI (not destroyed)
• 3′ cloning site: SpeI (not destroyed)
• 5′ sequencing primer: ATGGCGCCTCATCCCT
• 3′ sequencing primer: GTTC TCAGACTTCTTG

Gene/Insert 2

• Gene/Insert name: pdx1
• Species: M. musculus (mouse)
• Insert Size (bp): 852
• Entrez Gene: Pdx1 (a.k.a. IDX-1, IPF-1, Ipf1, Mody4, STF-1, pdx-1)

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: ATGAACAGTGAGGAG
• 3′ sequencing primer: CAGGAACCCCGG

Gene/Insert 3

• Gene/Insert name: Mafa
• Species: M. musculus (mouse)
• Insert Size (bp): 1077
• Entrez Gene: Mafa (a.k.a. RIPE3b1)

Cloning Information for Gene/Insert 3

• Cloning method: Restriction Enzyme
• 5′ cloning site: ClaI (not destroyed)
• 3′ cloning site: ClaI (not destroyed)
• 5′ sequencing primer: ATGGCCGCGGAGC
• 3′ sequencing primer: CACCCGACTTCTTTCTG

Gene/Insert 4

• Gene/Insert name: monomeric cherry
• Species: Synthetic
• Insert Size (bp): 711

Cloning Information for Gene/Insert 4

• Cloning method: Restriction Enzyme
• 5′ cloning site: NotI (not destroyed)
• 3′ cloning site: NotI (not destroyed)
• 5′ sequencing primer: atggtgagcaagggcgagg
• 3′ sequencing primer: ggacgagctgtacaagtaa

Resource Information

• Articles Citing this Plasmid:
  • 4 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

For more information, please see the associated article (Li, et al., Nat Biotechnol. 2014 PMID: 25402613.), as well as Li, et al., eLife. 2014, PMID: 24714494.

Note: The plasmid contains an N103Y amino acid residue substitution in the mouse Pdx1 gene. This residue change was present in the depositor's original plasmid stock as described in the associated publication and and should not affect plasmid function.