Regulatory
pLac

Part:BBa_K1638000:Design

Designed by: Jens Sivkr Pettersen   Group: iGEM15_SDU-Denmark   (2015-05-07)


Lac operator


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

This lac promoter with lac operator and CAP binding site is derived from either the pKT25 or pUT18C plasmid. In our case, this part was amplified alongside the T18 and T25 domain (the two catalytic active domains from the adenylate cyclase, CyaA, from Bordetella pertussis). For construction through PCR, we recommend the following primers:

Forward: 5'-ATATGAATCGCGGCCGCTTCTAGAGCTGGCACGACAGGTTTCCCG-3' - Contains EcoRI and XbaI restriction site

Reverse: 5'-ATATCTGCAGCGGCCGCTACTAGTATTGTTATCCGCTCACAATTCCACAC-3' - Contains SpeI and PstI restriction site

Source

pKT25, pUT18C

References