Composite

Part:BBa_K4131011:Design

Designed by: Chun Ying Yu   Group: iGEM22_CCU_Taiwan   (2022-10-10)


The lac-induced expressing cassette of pfa B, pfa C and pfa E (EPA)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 6107
    Illegal XbaI site found at 7363
    Illegal PstI site found at 3288
    Illegal PstI site found at 6044
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 6107
    Illegal NheI site found at 9562
    Illegal PstI site found at 3288
    Illegal PstI site found at 6044
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 6107
    Illegal BglII site found at 7855
    Illegal BamHI site found at 1542
    Illegal XhoI site found at 3983
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 6107
    Illegal XbaI site found at 7363
    Illegal PstI site found at 3288
    Illegal PstI site found at 6044
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 6107
    Illegal XbaI site found at 7363
    Illegal PstI site found at 3288
    Illegal PstI site found at 6044
    Illegal NgoMIV site found at 534
    Illegal AgeI site found at 6863
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 9525
    Illegal SapI site found at 1752


Design Notes

The pfa B (BBa_K4131008) and pfa E (BBa_K4131007) are codon optimized for biobrick assembly, while the pfa C is the original version in Moritella marina


Source

The lac promoter and operon (Bba_R0010) are located at the 5' terminal of this composite part to regulate the pfa B, pfa C, and pfa E genes transcription sequently. The t7 terminator (Bba_K3778018) is t the 3' terminal of this composite part.

The ribosome binding site (Bba_K3778013) is respectively added before pfa B, pfa C, and pfa E genes to mediated the individual protein translation.


References