pPAGFP-C1
(Plasmid #11910)

• Depositing Lab: Jennifer Lippincott-Schwartz
• Publication: Patterson et al Science. 2002 Sep 13. 297(5588):1873-7.

Backbone

• Vector backbone: pEGFP-C1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 4731
• 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: Unknown

Gene/Insert

• Gene/Insert name: PAGFP
• Alt name: photoactivatable green fluorescent protein
• Species: Aequorea victoria
• Mutation: These plasmids are essentially the same as the Clontech pEGFP-C1 version with four mutations (L64F/T65S/V163A/T203H). The PAGFP also contains an A206K mutation to disrupt dimerization

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: n/a

Resource Information

• Articles Citing this Plasmid:
  • 6 References

Terms and Licenses

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

Depositor Comments

The pPAGFP-C1 was not published. It was prepared by removing the PAGFP cDNA from pPAGFP-N1 with an AgeI/BsrGI restriction digest and subcloning it into a similarly digested pEYFP-C1 (Clontech). The pPAGFP-C1 was confirmed by sequencing. If you use this construct in future publications, use your best judgement for citation.

The fluorescent protein encoding region contains one of the three substitutions (A206K) suggested by Dr. Roger Tsien. This mutation disrupts fluorescent protein dimerization even at high concentrations. The mutation does not seem to have much effect on the fluorescent properties of the CFP, GFP and YFP versions, so hopefully it will not affect PAGFP too much. The PAGFP A206K mutant does not display obvious fluorescent differences compared with the original, but please bear in mind that I haven't characterized it to the same extent.

The citation for the A206K mutation in fluorescent proteins is Science 2002 296:913.

Information for Cloning Grade DNA (Catalog # 11910-DNA.cg) ( Back to top)

Purpose

Cloning grade DNA is suitable for use in PCR, cloning reactions, or transformation into E. coli. The purity and amount is not suitable for direct transfections.

Delivery

• Amount: 2 µg
• Guaranteed Concentration: 100 ng/µl +/- 5 ng/µl
• Pricing: $105 USD
• Storage: DNA can be stored at 4℃ (short term) or -20℃ (long term).

Terms and Licenses

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

Quality Control

Addgene has verified this plasmid using Next Generation Sequencing. Results are available here