Coding

Part:BBa_K3272005:Design

Designed by: Neval Sevinc   Group: iGEM19_ACIBADEM_ISTANBUL   (2019-10-16)


LTNF without stop codon in pPIC9K


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

The expression vector that we used last year, pPICZalphaA, contains zeocin as a selective marker. As we aim to manufacture our end product, we conclude that using zeocin antibiotic is not cost-effective. Instead, we used pPIC9K expression vector which contains kanamycin and ampicillin as selective markers, which are much more cheaper than zeocin. Both vectors contain AOX1 promoter enabling methanol-induced expression; and α-factor secretion signal to direct secreted expression of our protein.


Source

The coding region has the same sequence of LTNF 2.0 coding region that we submitted last year with part number BBa_K2815001; however it is now cloned in another vector, pPIC9K instead of pPICZalphaA that we used last year.

References