Regulatory
pLac
Part:BBa_K1638000:Design
Designed by: Jens Sivkr Pettersen Group: iGEM15_SDU-Denmark (2015-05-07)
Lac operator
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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