Plasmid_Backbone

Part:BBa_K5299007:Design

Designed by: Maria Nefeli Stoupa   Group: iGEM24_Thessaly   (2024-09-24)


pTN7-19[g1]: KmR, GmR, ori R6K, Tn7L and Tn7R extremes, pBBR1 (ori), 1AI2 cargo (for Level 1 reactio


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 56
    Illegal EcoRI site found at 333
    Illegal XbaI site found at 306
    Illegal XbaI site found at 709
    Illegal SpeI site found at 771
    Illegal PstI site found at 294
    Illegal PstI site found at 721
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 56
    Illegal EcoRI site found at 333
    Illegal SpeI site found at 771
    Illegal PstI site found at 294
    Illegal PstI site found at 721
    Illegal NotI site found at 46
    Illegal NotI site found at 739
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 56
    Illegal EcoRI site found at 333
    Illegal BglII site found at 1037
    Illegal BglII site found at 3232
    Illegal BamHI site found at 312
    Illegal BamHI site found at 703
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 56
    Illegal EcoRI site found at 333
    Illegal XbaI site found at 306
    Illegal XbaI site found at 709
    Illegal SpeI site found at 771
    Illegal PstI site found at 294
    Illegal PstI site found at 721
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 56
    Illegal EcoRI site found at 333
    Illegal XbaI site found at 306
    Illegal XbaI site found at 709
    Illegal SpeI site found at 771
    Illegal PstI site found at 294
    Illegal PstI site found at 721
    Illegal NgoMIV site found at 2271
    Illegal NgoMIV site found at 2275
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI site found at 678
    Illegal BsaI.rc site found at 94


Design Notes

R2K origin of replication requires a pir strain for replication


Source

We used PacI and SpeI restriction enzymes to excise the transcriptional unit from the pSEVA23g19[g1] vector and transfer it into the pTn7-M vector, following the method described by Sergio Salgado et al.

References