Translational_Unit

Part:BBa_K4158011:Design

Designed by: Nanami Onishi, Eri Ito   Group: iGEM22_Waseda_Tokyo   (2022-09-22)


RBS-GFP


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.rc site found at 658


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, this part and K4158013.


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


5'-GATAAGCTTCGAATTCCTAATCGTCGTAGATGGCGGAAACCGCGGCGGGATTCTGTTCGCATGCCCTGACAGCGGAAGTAGGAAAGCGGTCGTCGCATAACGCCCGCACGGAGCGCTGCAGGGATCCTCT-3'


3. Amplitude  K4158013 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 this part 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