Plasmid_Backbone

Part:BBa_K4435305:Design

Designed by: Elizabeth Bilsland   Group: iGEM22_Unicamp_Brazil   (2022-10-08)


level 2 JUMP plasmid, pBBR1 ori, AmpR


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found at 3060
    Illegal suffix found at 16
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3060
    Illegal SpeI site found at 17
    Illegal PstI site found at 31
    Illegal NotI site found at 24
    Illegal NotI site found at 3066
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3060
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 3060
    Illegal suffix found at 17
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 3060
    Plasmid lacks a suffix.
    Illegal XbaI site found at 3075
    Illegal SpeI site found at 17
    Illegal PstI site found at 31
    Illegal NgoMIV site found at 749
    Illegal NgoMIV site found at 878
    Illegal NgoMIV site found at 1373
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI site found at 3082
    Illegal BsaI.rc site found at 10


Design Notes

Validated by colony coloration, resistance to ampicillin, and loss of resistance to Kanamycin


Source

Amplification of BBa_J428367 with JUMP_mF and JUMP_mF; amplification of antibiotic resistance marker AmpR from BBa_J428385; Golden gate cloning with SapI. Primers: JUMP-mR GCTCTTCagactagagcgaagtcttcg JUMP-mF GCTCTTCTccttttccgctgcataacc AmpR_F GCTCTTCTCCTatttaaattagtagcccgcctaatgag AmpR_R GCTCTTCAGACgacaagggtcgtccaaaaaaaaagg

References