pCRT7 NT Topo tetR/pLtetO Amp‐WT sfGFP
(Plasmid #52053)

• Purpose: Used in phosphoprotein synthesis. Replace the sfGFP with gene of interest (GOI). Add TAG amber codon(s) at the position(s) in GOI where phosphoserine incorporation is desired.
• Depositing Lab: Jesse Rinehart
• Publication: Rinehart Lab Phosphoprotein synthesis components (unpublished)

Backbone

• Vector backbone: pCRT7/NT-TOPO
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 2870
• Total vector size (bp): 4279
• Modifications to backbone: tetR/pLtetO promoter system added
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: sfGFP
• Species: Synthetic
• Promoter: tetR/pLtetO

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: KpnI (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: Tet-R
• 3′ sequencing primer: T7 terminal

Resource Information

• Supplemental Documents:
  • Users Guide for Phosphoprotein reagents.pdf
• Articles Citing this Plasmid:
  • 4 References

Terms and Licenses

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

Depositor Comments

To clone new genes for your proteins of interest into the pCRT7 NT vector provided, use 5’KpnI--‐3’HindIII restriction sites.
See Lutz and Bujard 1997 (PMID 9092630) for information on the tetR/pLtetO promoter system used for inducible expression in the pCRT7 NT vector.
The phosphoproteins (or sfGFP) expressed via the pCRT7 pLtetO/tetR plasmid is induced with 100 ng/mL anhydrotetracycline.