Composite

Part:BBa_K4201019

Designed by: Lindsey Neukirch   Group: iGEM22_CU-Boulder   (2022-10-12)
Revision as of 10:27, 12 October 2022 by Dabirk (Talk | contribs)

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crtE-cytoTDS-MBP_T5αOH_RUBY

This composite part is made using a Kan BsaI backbone. The B/W selection was excised during Golden Gate synthesis and replaced with a sequence containing CrtE-cytoTDS-MBP, T5αOH, and RUBY genes. Each inserted gene is flanked by a Gmubi promoter and AtHSP terminator. This construct was made to be transfected into agrobacterium and then into Glycine max. The Crte-cytoTDS-MBP is a fusion protein that codes for the first two enzymes in the paclitaxel biosynthesis pathway, a GGPP synthase1 and taxadiene synthase2. The T5αOH encodes for the third step in the biosynthesis of paclitaxel3. The RUBY reporter was added to visualize the construct when integrated into the G. max genome4

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 6826
    Illegal PstI site found at 994
    Illegal PstI site found at 4792
    Illegal PstI site found at 5632
    Illegal PstI site found at 8365
    Illegal PstI site found at 13549
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 6826
    Illegal PstI site found at 994
    Illegal PstI site found at 4792
    Illegal PstI site found at 5632
    Illegal PstI site found at 8365
    Illegal PstI site found at 13549
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 6826
    Illegal BglII site found at 790
    Illegal BglII site found at 1610
    Illegal BglII site found at 3148
    Illegal BglII site found at 4195
    Illegal BglII site found at 5566
    Illegal BglII site found at 6493
    Illegal BglII site found at 7442
    Illegal BamHI site found at 5246
    Illegal BamHI site found at 11769
    Illegal XhoI site found at 2571
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 6826
    Illegal PstI site found at 994
    Illegal PstI site found at 4792
    Illegal PstI site found at 5632
    Illegal PstI site found at 8365
    Illegal PstI site found at 13549
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 6826
    Illegal PstI site found at 994
    Illegal PstI site found at 4792
    Illegal PstI site found at 5632
    Illegal PstI site found at 8365
    Illegal PstI site found at 13549
    Illegal NgoMIV site found at 6975
    Illegal NgoMIV site found at 9705
    Illegal NgoMIV site found at 10284
    Illegal NgoMIV site found at 11997
    Illegal AgeI site found at 937
    Illegal AgeI site found at 2313
    Illegal AgeI site found at 5429
    Illegal AgeI site found at 5701
    Illegal AgeI site found at 6640
    Illegal AgeI site found at 8162
    Illegal AgeI site found at 8434
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1
    Illegal BsaI site found at 940
    Illegal BsaI site found at 5432
    Illegal BsaI site found at 5704
    Illegal BsaI site found at 6643
    Illegal BsaI site found at 8165
    Illegal BsaI site found at 8437
    Illegal BsaI.rc site found at 934
    Illegal BsaI.rc site found at 5426
    Illegal BsaI.rc site found at 5698
    Illegal BsaI.rc site found at 6637
    Illegal BsaI.rc site found at 8159
    Illegal BsaI.rc site found at 8431
    Illegal BsaI.rc site found at 9370
    Illegal SapI.rc site found at 3369
    Illegal SapI.rc site found at 3575


[edit]
Categories
Parameters
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