Plasmid

Part:BBa_K3579023

Designed by: Amandine Maire   Group: iGEM20_Paris_Bettencourt   (2020-10-27)


EpiFlex_pPBEF1_mCherry_AB

This plasmid is a level 1 plasmid containing the TU coding for mCherry and assembled using the EpiFlex toolkit (MoClo for s. epidermidis). The TU is composed of a SarA promoter, a SodA RBS, a mCherry CDS and a T7 terminator. This level 1 plasmid is designed with level 2 fusion sites A and B which aims to place it in position 1 in the genetic circuit assembled in the level 2 plasmid.

This plasmid has a pPBEF_bb1 backbone with kanamycin resistance gene for E. Coli and erythromycin resistance gene for S. epidermidis. The plasmid is a E. coli -> S. epidermidis shuttle vectors. This means that they contain origins of replication for both E. coli and S. epidermidis.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 3509
    Illegal SpeI site found at 1682
    Illegal PstI site found at 1812
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 3509
    Illegal NheI site found at 7740
    Illegal SpeI site found at 1682
    Illegal PstI site found at 1812
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 3509
    Illegal BglII site found at 1539
    Illegal BglII site found at 1621
    Illegal BamHI site found at 207
    Illegal BamHI site found at 1667
    Illegal XhoI site found at 1660
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 3509
    Illegal SpeI site found at 1682
    Illegal PstI site found at 1812
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 3509
    Illegal SpeI site found at 1682
    Illegal PstI site found at 1812
    Illegal NgoMIV site found at 7888
    Illegal AgeI site found at 809
    Illegal AgeI site found at 7357
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 13
    Illegal BsaI site found at 103
    Illegal BsaI site found at 175
    Illegal BsaI site found at 994
    Illegal BsaI site found at 8007
    Illegal BsaI.rc site found at 65
    Illegal BsaI.rc site found at 137
    Illegal BsaI.rc site found at 956
    Illegal BsaI.rc site found at 1105
    Illegal BsaI.rc site found at 7154
    Illegal SapI site found at 4265
    Illegal SapI site found at 5041


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