Composite

Part:BBa_K3268002:Design

Designed by: YI-HSUAN CHAN   Group: iGEM19_NYMU-Taipei   (2019-10-20)


Strong expression of eforRed in E. coli


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 2047
    Illegal PstI site found at 14
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2047
    Illegal NheI site found at 2075
    Illegal NheI site found at 2098
    Illegal PstI site found at 14
    Illegal NotI site found at 7
    Illegal NotI site found at 2053
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2047
    Illegal BamHI site found at 2124
    Illegal XhoI site found at 1031
    Illegal XhoI site found at 1923
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 2047
    Illegal PstI site found at 14
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 2047
    Illegal XbaI site found at 2062
    Illegal PstI site found at 14
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

1. In order to make the strong promoter (along with BBa_B0034 RBS and BBa_B0015 transcriptional terminator on pSB1C3 plasmid backbone) useful to highly express inserted protein-coding genes, we had designed two PCR primers with HindII and BamHI sites respectively to generate a composite expression module without the original GFP coding sequence. 2. Any inserted protein-coding genes not containing HindII and BamHI sites in their coding sequences can then be amplified and cloned into our composite expression module improved from BBa_J364007.


Source

1. The eforRed coding region was PCR amplified from BBa_K592012 using high-fidelity enzyme KOD-Plus. 2. The strong promoter along with BBa_B0034 RBS and BBa_B0015 transcriptional terminator on pSB1C3 plasmid backbone were PCR amplified from BBa_J364007 using high-fidelity enzyme KOD-Plus.

References