pYIC
(Plasmid #18673)

• Purpose: (Empty Backbone)
• Depositing Lab: Han Htun
• Publication: Nie et al Nucleic Acids Res. 2006 . 34(19):5528-40.

Backbone

• Vector backbone: pIRES2-EGFP
• Backbone manufacturer: Clontech
• Backbone size (bp): 5300
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Bicistronic fluorescent reporter gene with cap-dependent EYFP and IRES-dependent ECFP translation under the control of CMV promo
• Species: H. sapiens (human); Aequorea victoria; synthetic sequence.
• Insert Size (bp): 895
• Tags / Fusion Proteins:
  • 3Myc-EYFP-HA-His6 (N terminal on insert)
  • ECFP-HA-His6 (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SacI and BstXI (not destroyed)
• 3′ cloning site: Eco RI and Not I (not destroyed)
• 5′ sequencing primer: CMV-F

Resource Information

• Supplemental Documents:
  • pYIC
• A portion of this plasmid was derived from a plasmid made by: ECFP and EYFP were PCR amplification products from pECFP-Mito and pEYFP-N1 DNAs from Clontech.
• Articles Citing this Plasmid:
  • 9 References

Terms and Licenses

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

Depositor Comments

EYFP tagged at N-terminus with 3 copies of Myc epitope tags and at C-terminus with a copy of FLAG and polyhistidine (His6) epitope tags. ECFP tagged at the C-terminus with one copy of hemagglutinin (HA) and polyhistidine (His6) epitope tags.

EGFP of pIERS2-EGFP (Clontech) replaced at an 803 bp BstXI-NotI, PCR amplified fragment with ECFP from pECFP-Mito (Clontech) tagged at the C-terminus with a single copy of hemagglutinin (HA) and polyhistidine (His6) epitope tags; IRES-dependent translation yields ECFP-HA-His6 fusion protein. 895 bp Sac I/Eco RI fragment containing amino terminal 3 Myc epitope tags fused to EYFP obtained from PCR amplification of pEYFP-N1 (Clontech), which is also tagged at C-terminus with a single copy of FLAG and polyhistidine (His6) epitope tag; cap-dependent translation yields 3Myc-EYFP-Flag-His6 fusion protein.

See article for detailed information.