Coding

Part:BBa_K1590004:Design

Designed by: Manuel Blank   Group: iGEM15_Dundee   (2015-09-09)


Regulator or Chromate responsive promoter


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

The part was partially optimised by altering the first 15 codons by means of a non-specific primer. The codons were biased towards maximum expression level in E. coli K12 strains.

Primers used for amplification of ChrB coding-region, while optimising the 15 codons, adding a standard prefix, and a standard suffix to it: 

  Forward: GCGC GAATTCGCGGCCGCTTCTAG A TGAACCTGCTGTCTCCGATCCTGTCTCTGCCGACCGAAAACGCG

  Reverse: GCGC CTGCAGCGGCCGCTACTAGT ATTATT ATACGGTGAGGGTCCC

Primers for amplification of partially optimised ChrB from BBa_K1058008 and adding a standard prefix, a PstI restriction site, and a His6-tag to it. 

  Forward: GCGC GAATTCGCGGCCGCTTCTAG A TGAACCTGCTGTCTCCGATCCTGTCTCTGCCGACCGAAAACGCG

  Reverse: GCGC CTGCAG TTATTA ATGGTGATGGTGATGGTGTACGGTGAGGGTCCC

Primers for amplification of partially optimised ChrB from recombinant plasmid and adding BamHI/PstI restriction sites and a His6-tag to it. 

  Forward: GCGC GGATCC ATGAACCTGCTGTCTCCGATCCTGTCT

  Reverse: GCGC CTGCAG TTATTA ATGGTGATGGTGATGGTG TACGGTGAGGGTCCC


Source

PCR-amplification from biobrick BBa_K1058008 (pChrBGFP).

References