Plasmid_Backbone

Part:BBa_J176171:Design

Designed by: Karmella Haynes   Group: Haynes Lab   (2013-04-19)


Kozak-XbaI-NLS-6His-stop


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 266
    Illegal XbaI site found at 329
    Illegal SpeI site found at 249
    Illegal PstI site found at 271
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 266
    Illegal SpeI site found at 249
    Illegal PstI site found at 271
    Illegal NotI site found at 292
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 266
    Illegal BglII site found at 12
    Illegal XhoI site found at 299
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 266
    Illegal XbaI site found at 329
    Illegal SpeI site found at 249
    Illegal PstI site found at 271
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 266
    Illegal XbaI site found at 329
    Illegal SpeI site found at 249
    Illegal PstI site found at 271
    Illegal NgoMIV site found at 819
    Illegal NgoMIV site found at 2230
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI.rc site found at 1955
    Illegal BsaI.rc site found at 3755
    Illegal SapI site found at 2675


Design Notes

  • A Kozak-XbaI-NLS-6His-stop with SpeI sticky overhangs was inserted into the XbaI site.
  • Sequencing data confirms proper orientation of the inserted part.
  • XbaI site can accept any Silver Standard (RFC 23) protein coding region that is cut with XbaI/SpeI.
  • Single-cut vectors can easily re-ligate: It is highly recommended that the user dephosphorylates the XbaI-cut vector prior to ligation.
  • Insertion of a XbaI/SpeI insert is non-directional: It is high recommended that the user selects at least 3 colonies after ligation check (vi sequencing or some other method) for proper orientation of the insert.


Source

Built from BBa_J176122

References