Plasmid_Backbone

Part:BBa_K2656200:Design

Designed by: Carolina Ropero   Group: iGEM18_Valencia_UPV   (2018-09-21)


pBIOBRICKATOR


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found at 5
    Plasmid lacks a suffix.
    Illegal SpeI site found at 921
    Illegal PstI site found at 935
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 5
    Illegal NheI site found at 44
    Illegal NheI site found at 67
    Illegal SpeI site found at 921
    Illegal PstI site found at 935
    Illegal NotI site found at 11
    Illegal NotI site found at 928
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 5
    Illegal XhoI site found at 1952
    Illegal XhoI site found at 2844
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 5
    Illegal suffix found at 921
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 5
    Plasmid lacks a suffix.
    Illegal XbaI site found at 20
    Illegal SpeI site found at 921
    Illegal PstI site found at 935
    Illegal AgeI site found at 653
    Illegal AgeI site found at 765
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.


Design Notes

pBioBrickator plasmid was constructed by using a Gibson assembly of the RFP gene and pUD2 backbone. BsmBI recognition sites and GB 3.0 BsmBI alpha 1 overhangs (5’-ggagtgagacg-3’ and 5’-cgtctcagtca-3’) where inserted into this new plasmid by adding them between the template complementary sequence and the homologous overlapping end of each primer. Thus, we have used this primers: ctggaattcgcggccgcttctagagggagtgagacgtttacggctagctc and tttctgcgtttatacgtctcagtcaactagtagcggccgctgcagtc to amplificate the pBioBrickator backbone; and ggccgcttctagagggagtgagacgtttacggctagctcagtcctaggta and ggtgggcctttctgcgtttatacgtctcagtcaactagtagcggccg to amplificate the red transcriptional unit that acts like screening marker.


Source

The original sequence has been obtained from the Registry of Standard Biological Parts. It has been modified using the Gibson assembly method.

References