Coding

Part:BBa_K2194003:Design

Designed by: Catherine Dunaway, Anna Guseva   Group: iGEM17_Rice   (2017-10-23)
Revision as of 19:37, 1 November 2017 by Catdunaway (Talk | contribs) (→‎Design Notes)


cysPUWA Sulfate Transport System cds


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1274
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 2090
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 3
    Illegal BsaI.rc site found at 3828


Design Notes

The entire cysPUWA sequence was too long to manufacture as one gBlock, so we split the sequence into cysPU and cysWA and added BsmBI sites with complementary sticky ends to the end of cysPU and the beginning of cysWA so that the two gBlocks could be combined using a scarless Golden Gate Assembly reaction.

Source

We sourced the nucleotide sequence for cysPUWA from the Escherichia coli MG1655 genome NCBI database (entry NC_000913.3:2539714-2543526). We then sent this sequence to IDT to manufacture as two gBlocks®.

CysPUWA cds NCBI.png

Figure 1: This image is taken from NCBI and shows the cysPUWA genes in their genomic context.

References

[update]