Plasmid

Part:BBa_K2141000

Designed by: Altug Uludag, Nedim Haciosmanoglu, Galip Can Guclu, Yagmur Guneri, Melis Akgun   Group: iGEM16_Istanbul_Tech   (2016-10-04)


pET28a(+) expression plasmid

pET28a(+) is a nonviral expression plasmid used in bacteria strains for protein production purposes. It has kanamycin resistance gene as a selection marker. Plasmid sequence consists of T7 promoter and terminator sequences as well as lac operon sequence and lacI promoter for protein expression induction. It has both N- and C- terminal HisTag sequences and N- terminal thrombin cleavage site.
<be>

Additional Information

Team: IISERPUNE-INDIA-2020

  • Pet28a(+) is widely used for expression purpose. But unnecessary components often interfere in cloning strategies. So we altered the part into a modified version that satisfies RFC10 criteria, compatible with Gibson's cloning and all the unnecessary components removed: BBa_K358500.


Please see the experience page for more information.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 192
    Illegal XbaI site found at 335
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 192
    Illegal NheI site found at 231
    Illegal NotI site found at 165
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 192
    Illegal BglII site found at 401
    Illegal BamHI site found at 198
    Illegal XhoI site found at 158
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 192
    Illegal XbaI site found at 335
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 192
    Illegal XbaI site found at 335
    Illegal NgoMIV site found at 433
    Illegal NgoMIV site found at 2021
    Illegal NgoMIV site found at 2181
    Illegal NgoMIV site found at 5228
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 3101


[edit]
Categories
Parameters
None