pcDNA3 GFP LIC cloning vector (6D)
(Plasmid #30127)

• Purpose: (Empty Backbone)
• Depositing Lab: Scott Gradia
• Publication: 6-series mammalian LIC expression vectors (unpublished)

Backbone

• Vector backbone: pcDNA3
• Backbone size (bp): 6145
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

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: None
• Tag / Fusion Protein:
  • GFP (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: LIC site vKoz/GFP (destroyed during cloning)
• 3′ cloning site: LIC site vKoz/GFP (destroyed during cloning)
• 5′ sequencing primer: CMV F (5'cgcaaatgggcggtaggcgtg)
• 3′ sequencing primer: GFP reverse (5'cagctcgaccaggatgggc3')

Resource Information

• Articles Citing this Plasmid:
  • 10 References

Terms and Licenses

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

Depositor Comments

This is an empty LIC vector derived from pcDNA3. It adds a GFP gene to the C-terminus of your protein of interest.

GFP has a excitation max of 489 nm and an emission max of 510 nm.

To clone into this vector, add the following tags to your PCR primers:

LIC vKoz Forward tag 5’-TACTTCCAATCCAATGCCACC(ATG)

LIC vGFP Reverse tag 5’-CTCCCACTACCAATGCC

Do NOT include a stop codon with your reverse primer.

T4-treat PCR with dCTP. Linearize vector with SspI, then T4-treat with dGTP. Can verify the presence of insert by digesting with HindIII and XbaI.

For more information, please see our website:
http://qb3.berkeley.edu/qb3/macrolab/