DNA

Part:BBa_K4630101:Design

Designed by: Zhejun Qin   Group: iGEM23_WHU-China   (2023-10-10)


stgRNA-barcode-cassette 2


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

Using Biobrick Assembly, the part can be inserted downstream of stgRNA-barcode-cassette 1, allowing the device to be knocked out twice. Also, the barcode is different from stgRNA-barcode-cassette 1, to ensure recombination uniqueness. Furthermore, a universal RNA linker was designed between the leading barcode and the stgRNA, ensuring the RNA functionality.

For the convenience of inserting stgRNA barcode cassette 2 into the plasmid vector, we inserted BamHI and EcoRI restriction enzyme sites on both sides of the fragment, respectively.In addition, we use the biobrick method to connect cassettes, so we have added SpeI cleavage sites after the fragments to facilitate the insertion of the next level of cassette.

Design Graph

                <img style="margin:20px auto 5px auto;" src="5-6x.png" width="20%">

Fig 1 Design Graph of stgRNA-barcode-cassette 2

Source

Complete synthesis.

References