PBP43

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pBP43: click to enlarge

General comments

Integrative plasmid for the fusion of the monomeric variant of GFP A206K to the C-terminus of a protein, keeping the control of the expression under its natural promoter

  • E.coli: multicopy plasmid, select for ampicillin
  • B. subtilis: no ori, select for spectinomycin

Hints for the usage

  • amplify the last 600bp of your gene of interest,
  • skip the Stopp-Codon of the gene in the reverse primer
  • take care that the integration is in-frame
  • do not use the HindIII site for cloning, as GFP is inserted via this restriction site
  • Don't use XbaI and KpnI for cloning. They cut the vector twice!

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