Regulatory

Part:BBa_K4158013:Design

Designed by: Nanami Onishi, Eri Ito   Group: iGEM22_Waseda_Tokyo   (2022-09-25)


PedcR


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 145
    Illegal SapI site found at 9
    Illegal SapI site found at 37


Design Notes

If you want to get the same plasmid that we experimented with, construct the plasmid along this way below.


1. Prepare pACYC184 cloning vector, BBa_K3146011 and this part.


2. Restriction and insertion cloning with pACYC184, DNA Flagment Below, and restriction enzymes HindIII , BamHI.


5'-GATAAGCTTCGAATTCCTAATCGTCGTAGATGGCGGAAACCGCGGCGGGATTCTGTTCGCATGCCCTGACAGCGGAAGTAGGAAAGCGGTCGTCGCATAACGCCCGCACGGAGCGCTGCAGGGATCCTCT-3'


3. Amplitude  this parts with PCR using These Primers 


Fw: 5'-GCTGCAGCGCGTCCTGCTCTTCC-3'


Rv: 5'-GTAGAGGATCCGTCGACAAAAAACGCACTT-3'


4. Restriction and insertion cloning with the PCR productions and insertion cloning (Process 2) products and restriction enzymes PstI and BamHI.


5. Amplitude K4158011 with PCR using These Primers 


Fw: 5'-TGTCGACCTGCAGGCATGCAAGCTTAGGAGGAAAAACATATGAGTAAA-3'


Rv: 5'-AGGATCCACTAGTTCTATACGCGAAATAGCAGAC-3'


6. Restriction and insertion cloning with the PCR productions and insertion cloning (Process 4) products and restriction enzymes SalI and BamHI.

Source

test

References