Regulatory
Part:BBa_K1104204:Design
Designed by: Ting-Yun Chiang Group: iGEM13_NYMU-Taipei (2013-09-16)
AhpCp2D1
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
We delete the dsbG coding and following sequence next to dsbG reverse primer, making the functional sequence shorter, so that PCR can be more easy and efficient. The length of remained parts is about half of the original part: K362001.
Besides, AhpCp2D1 has a Pst1 site in the sequence. We succesfully mutated the Pst1 site and make Assembly Compatibility better.
Primers For PCR
| AhpCp2_DsbGp_AhpCp_FP | gcttctagag caatcgcttttactggagcaggaa |
| AhpCp_RP | gga ctgcagcggccgctactagta ctatacttcctccgtgttttcgatgaga |
| ahpC promoter(K362001)has a Pst1 site in the sequence. We designed this part AhpCp2D1, and succesfully mutated the Pst1 site (ctgcag->ctacag). | |
|---|---|
| ahpCp_mut_FP | cacctcagcctacaggcaggcactgaa |
| ahpCp_mut_RP | ttcagtgcctgcctgtaggctgaggtg |
PCR Time Protocol
| Temperature(°C) | Time(sec) | Number of Cycles |
|---|---|---|
| 94 | 120 | -- |
| 94 | 15 | 35 |
| 53 | 30 | |
| 72 | 30 | |
| 72 | 300 | -- |
Source
E. coli MG1655 genomic sequence