Regulatory

Part:BBa_K1104204:Design

Designed by: Ting-Yun Chiang   Group: iGEM13_NYMU-Taipei   (2013-09-16)

AhpCp2D1


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

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

References