Plasmid_Backbone

Part:BBa_K4133000:Design

Designed by: Luis Alejandro Chaves Martinez   Group: iGEM22_Costa_Rica   (2022-10-02)


pLBov plasmid (Lactobacillus/E. coli shuttle vector)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 4975
    Illegal NheI site found at 2290
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 4981
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 4975
    Illegal BglII site found at 2892
    Illegal XhoI site found at 3959
    Illegal XhoI site found at 4851
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 4975
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 4975
    Plasmid lacks a suffix.
    Illegal XbaI site found at 4990
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.


Design Notes

Due to the cost issues related to vector synthesis, pLBov was synthesized in fragments and assembled using Single-Pot Scarless Golden Gate assembly with the following overhangs:


Fragmentation of pLBov for assembly by Single-Pot Scarless Golden Gate assembly.
pLBov fragment Prefix Suffix Location (nt)
1 GGTCTC A CAGT AACT T GAGACC
2 GGTCTC T AACT TCGT C GAGACC
3 GGTCTC G TCGT TAGG C GAGACC
4 GGTCTC A TAGG CAGT C GAGACC

Source

This plasmid was obtained by de-novo synthesis.