Plasmid

Part:BBa_K3776020

Designed by: Helene Heling Huang   Group: iGEM21_HK_SSC   (2021-10-01)


panS-SPIN-davBA-AK

This part allows genome insertion of genes enabling induced lysine synthesis and 5-aminovalerate (5AVA) synthesis in Synechococcus elongatus UTEX 2973. The INTEGRATE system ( BBa_K3776019) that insert three genes as a donor DNA into the genome of S.elongatus UTEX 2973 is cloned into the panS-loxP-MCS shuttle vector ( BBa_K3776009). The shuttle vector can be replicated in E.coli for plasmid miniprep and in S.elongatus UTEX 2973 for expression. BBa_K3776006, consisting a feedback insensitive aspartate kinase ( BBa_K3776005) for overproduction of lysine, DavB ( BBa_K3776001) and DavA ( BBa_K3776002) for synthesis of 5AVA, has been cloned as the donor DNA in the INTEGRATE system.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7311
    Illegal NheI site found at 9398
    Illegal NheI site found at 13516
    Illegal NotI site found at 33
    Illegal NotI site found at 15236
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 206
    Illegal BglII site found at 6737
    Illegal BglII site found at 7528
    Illegal BglII site found at 8058
    Illegal BglII site found at 13303
    Illegal BglII site found at 16325
    Illegal BglII site found at 19280
    Illegal BamHI site found at 6
    Illegal BamHI site found at 5880
    Illegal BamHI site found at 14431
    Illegal XhoI site found at 5663
    Illegal XhoI site found at 14414
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2378
    Illegal NgoMIV site found at 16485
    Illegal NgoMIV site found at 18252
    Illegal AgeI site found at 4103
    Illegal AgeI site found at 13435
    Illegal AgeI site found at 15892
    Illegal AgeI site found at 16141
    Illegal AgeI site found at 17476
    Illegal AgeI site found at 19479
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 5846
    Illegal BsaI.rc site found at 5820
    Illegal SapI.rc site found at 8814


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Categories
Parameters
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