Difference between revisions of "Part:BBa K3794000:Design"

 
 
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===Design Notes===
 
===Design Notes===
−
Removed signal sequence from UniProt U5Y3S6. Codon optimised for expression in E.Coli K12  
+
Removed signal sequence from UniProt U5Y3S6. Codon optimised for expression in E.Coli K12.
 +
 
 +
Added N-terminal 6xHis tag with start codon and TEV cleavage site for protein purification via Ni-NTA column
 +
 
 +
Upstream of 6xHis tag, a start codon (ATG) was added to allow for transcriptional initiation, and a stop codon was added at 3' end of PVFP-5 CDS to allow for translational termination during protein synthesis
 +
 
  
  
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===Source===
 
===Source===
  
−
This part was synthesised by IDT. The DNA sequence was obtained from UniProtKB: U5Y3S6, and codon optimised for expression in E.Coli
+
This part was synthesised as a composite by IDT (with an upstream LacI regulatory (BBa_R0010). The protein sequence was obtained from UniProtKB: U5Y3S6, a 6xHis tag and TEV cleavage site was added, and codon optimised for expression in E.Coli K12
  
 
===References===
 
===References===

Latest revision as of 17:01, 21 October 2021


PVFP-5


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Removed signal sequence from UniProt U5Y3S6. Codon optimised for expression in E.Coli K12.

Added N-terminal 6xHis tag with start codon and TEV cleavage site for protein purification via Ni-NTA column

Upstream of 6xHis tag, a start codon (ATG) was added to allow for transcriptional initiation, and a stop codon was added at 3' end of PVFP-5 CDS to allow for translational termination during protein synthesis



Source

This part was synthesised as a composite by IDT (with an upstream LacI regulatory (BBa_R0010). The protein sequence was obtained from UniProtKB: U5Y3S6, a 6xHis tag and TEV cleavage site was added, and codon optimised for expression in E.Coli K12

References