Plasmid

Part:BBa_K4357004

Designed by: Di Yu   Group: iGEM22_UM_Macau   (2022-10-12)


Pasr-mCherry-pSB1C3

In the direction for our gene expression there is an asr promotor (BBa_K1231000) which is the pH-responsive promoter native to E. coli, inducing transcription in acidic conditions (~pH 5.5), RBS (BBa_B0034) which is the ribosome binding site,mCherry gene which is our fluorescent reporter gene, 6xhis which is our his tag, double terminator (BBa_B0015) which is the terminator, BioBrick suffix and his Oberon terminator. In the opposite direction, the cat promotor controls the CmR gene which is the selective marker, carrying chloramphenicol resistance., followed by Iambda t0 terminator. There is also a replication origin, pUC19-derived pMB1 (copy number of 100-300 per cell).

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 3110
    Illegal suffix found in sequence at 1062
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 3110
    Illegal SpeI site found at 1063
    Illegal PstI site found at 1077
    Illegal NotI site found at 881
    Illegal NotI site found at 1070
    Illegal NotI site found at 3116
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 3110
    Illegal XhoI site found at 890
    Illegal XhoI site found at 2094
    Illegal XhoI site found at 2986
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 3110
    Illegal suffix found in sequence at 1063
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 3110
    Illegal XbaI site found at 3125
    Illegal SpeI site found at 1063
    Illegal PstI site found at 1077
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
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