Plasmid

Part:BBa_K4716007

Designed by: LIAO WENYU   Group: iGEM23_UM-Macau   (2023-10-10)


pSB1C3-Pasr-CsgA-Perpa-mefp5-mSandy

On the direction for our gene expression, there is an Pasr promoter (BBa_K4716000) which is the pH-responsive promoter native to E.coli, inducing transcription in human duodenum region’s relatively acidic environment (pH 5~6), RBS (BBa_B0034) is the ribosome binding site, CsgA-Perpa-mafp-5 is our terget gene, the mSandy2 is the reporter gene and the double terminator (BBa_B0015) is the terminater we use in this circuit, BioBrick prefix and BioBrick suffix are presented at the beginning and end of the whole insert gene, respectively. In the opposite direction, the cat promoter controls the CmR gene which is the selective marker, carrying chloramphenicaol resistance gene for our screening object. Followed by Lambda T0 terminator, there is also a replication origin for the whole plasmid.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 2049
    Illegal suffix found in sequence at 1
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2049
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 2055
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2049
    Illegal BamHI site found at 3661
    Illegal XhoI site found at 1033
    Illegal XhoI site found at 1925
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 2049
    Illegal suffix found in sequence at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 2049
    Illegal XbaI site found at 2064
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None