Plasmid

Part:BBa_K4113022

Designed by: Wenhui Ren   Group: iGEM22_FAFU-China   (2022-10-07)


pCOMBIA1301SN-DlGAD2

The constructed recombinant plasmid pCOMBIA1301SN was used as a template for PCR using specific primers for the DlGAD2 gene of Longan. The results were shown in Figure 1, and the size of the PCR amplification product was about 1428 bp, which was consistent with the size of the DlGAD2 gene of Longan. To further verify the accuracy of the insertion site of DlGAD2 gene in Longan, the recombinant plasmid pCOMBIA1301- DlGAD2 was verified by double digestion using restriction endonucleases FastDigest KpnⅠ and FastDigest SalⅠ, and the digested product was imaged by electrophoresis gel, as shown in Figure 2, where the serial number ② represents the DlGAD2 after double digestion which length is 1428bp , and serial number ④ represents the band of pCOMBIA1301SN after double digestion by restriction endonucleases FastDigest KpnⅠ and FastDigest SalⅠ, indicating the correct insertion site of DlGAD2 on the vector. The combined results of PCR and double digestion showed that the DlGAD2 gene was successfully inserted into the polyclonal site of the intermediate expression vector pCOMBIA1301SN, which can be used for subsequent studies.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2018
    Illegal NheI site found at 5462
    Illegal NheI site found at 14401
    Illegal NotI site found at 3337
    Illegal NotI site found at 4869
    Illegal NotI site found at 6159
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 13224
    Illegal BamHI site found at 11582
    Illegal BamHI site found at 13537
    Illegal XhoI site found at 8905
    Illegal XhoI site found at 9999
    Illegal XhoI site found at 11587
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2782
    Illegal NgoMIV site found at 2956
    Illegal NgoMIV site found at 3006
    Illegal NgoMIV site found at 3010
    Illegal NgoMIV site found at 3037
    Illegal NgoMIV site found at 3536
    Illegal NgoMIV site found at 3894
    Illegal AgeI site found at 5956
    Illegal AgeI site found at 9626
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 5859
    Illegal BsaI site found at 14573
    Illegal SapI site found at 6550
    Illegal SapI.rc site found at 6087


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Categories
Parameters
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