pGH00.0126
(Plasmid #64257)

• Purpose: Binary T-DNA vector expressing EGFP and neomycin phosphotransferase II (NPTII); for Agrobacterium tumefaciens-mediated transformation
• Depositing Labs: Mark Guiltinan, Siela Maximova
• Publication: Maximova et al Plant Cell Rep. 2003 Jun;21(9):872-83. Epub 2003 Apr 3.

Backbone

• Vector backbone: pBin19 transformation vector
• Backbone manufacturer: Bevan M (1984) Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res 12:8711–8721
• Total vector size (bp): 15608
• Vector type: Plant Expression ; Binary Agrobacterium vector for plant transformation.

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: EGFP
• Promoter: E12-omega

Cloning Information for Gene/Insert 1

• Cloning method: Unknown
• 5′ sequencing primer: N/A

Gene/Insert 2

• Gene/Insert name: neomycin phosphotransferase II
• Alt name: NPTII
• Promoter: E12-omega

Cloning Information for Gene/Insert 2

• Cloning method: Unknown
• 5′ sequencing primer: N/A

Resource Information

• Supplemental Documents:
  • pGH00.0126 genbank file
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

E12-omega is a strong constitutive promoter obtained from pE2113-GUS (Mitsuhara et al. 1996). The promoter consists of two directly repeated copies the 5’ upstream sequence of the cauliflower mosaic virus (CaMV) 35S promoter (-419 to -90), followed by one copy of the P35S, the 5’-upstream sequence of the CaMV 35S promoter (–90 to –1) and the 5’ Omega-untranslated sequence of the tobacco mosaic virus. The Omega sequence is known to enhance translation levels in plant, animal and E. coli. This chimeric promoter was shown to support very high protein expression levels (approx. 23-fold higher then the standard CaMV 35S promoter) based on high transcription and enhanced translation rates (Mitsuhara et al. 1996).

Note-there is a 2nd NPTII cassette under the NOS Promoter. Only 1 NPTII region mutation was corrected and that is noted in the GenBank record.