Temporary

Part:BBa_K1484316

Designed by: Salil Bhate   Group: iGEM14_Cambridge-JIC   (2014-10-08)

Mp Selection Cassette

Marchantia selection cassette, level 1 part compatible with Modular Cloning.


Usage and Biology

This cassette confers 10ug/ml Hygromycin resistance to transformed mutants. It features the [Mp] driving the [HygR] and is terminated by [nos]

Use: If using A. tumefaciens mediated transformation, it is recommended that this part be adjacent to the Left Border of the T-DNA. Partial integrations can occur in transformation, and the Left Border is inserted last. Thus, if inserted adjacent to the Left Border, only those plants with complete integration of the T-DNA will be selected for.

This part has for this reason been designed to be the first part accepted into a Modular Cloning level 1 acceptor.

Time Frames: Transformed plantlets can be isolated approximately after 7 days' incubation with constant light, when the procedure below is followed.

Conditions: 

For effective selection of transformants after AT transformation, it is recommended to plate spores cocultivating with A. tumefaciens on 1/2 Gamborg's B5 + 1.2% Agar + 100ug/ml cefotaxime + 10ug/ml hygromycin following a wash with 150ml of sterile water containing 100ug/ml cefotaxime. Incubate at 22deg with constant light.

The selective media will not preclude fungal contimation, and it is thus important that all procedures are completed in sterile conditions. Micropore tape to seal plates is strongly recommended.

For further transformation information, visit:

or see the following literature resources:

Sequence and Features

Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 147
    Illegal BglII site found at 235
    Illegal XhoI site found at 1480
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 859
    Illegal NgoMIV site found at 923
    Illegal NgoMIV site found at 1065
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//chassis/eukaryote/mpolymorpha
Parameters
None