Difference between revisions of "Part:BBa K382002"

Revision as of 17:55, 5 November 2010

pORE Expression Series Vector with BioBrick MCS, Basta resistance

Vector for transformation of plants. Cloning in E. Coli, transfer to agrobacteria, integrates in the plant genome. Kanamycin resistance in bacteria, confers Basta resistance in plants. Contains the pENTCUP2 promoter upstream of MCS for constitutive expression of gene of interest.

Confirmed functions:

Kan resistance in bacteria: confirmed by transformation and colony growth of E. coli (DH5alpha) and Agrobacterium tumefaciens
Basta resistance in plants: confirmed by selective germination of transformed Arabidopsis seeds on basta agar plates (See photos [http://2010.igem.org/Team:Harvard/results]).

Confirmation pending: Function of the ENTCUP2 promoter will be tested by checking for expression for several BioBrick parts in adult Arabidopsis plants. These parts include...

miraculin (BBa_K382021)
brazzein (BBa_K382052)
GFP knock-down amiRNA (BBa_K382054)

Successfully transfected basta resistant Arabidopsis plants, such as the one shown below (brazzein BBa_K382052), will be tested for BioBrick expression once they have grown large enough for tissue sampling:

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 5326
Illegal XbaI site found at 5299
Illegal SpeI site found at 5378
Illegal PstI site found at 5356
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 5326
Illegal NheI site found at 4766
Illegal SpeI site found at 5378
Illegal PstI site found at 5356
Illegal NotI site found at 5341
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 5326
Illegal BglII site found at 5212
Illegal BglII site found at 7771
Illegal BamHI site found at 5312
Illegal XhoI site found at 4790
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 5326
Illegal XbaI site found at 5299
Illegal SpeI site found at 5378
Illegal PstI site found at 5356
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 5326
Illegal XbaI site found at 5299
Illegal SpeI site found at 5378
Illegal PstI site found at 5356
Illegal NgoMIV site found at 625
Illegal NgoMIV site found at 749
Illegal NgoMIV site found at 2181
Illegal NgoMIV site found at 2772
Illegal NgoMIV site found at 7220
Illegal AgeI site found at 341
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 7241

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K382002 short</partinfo>
-Vector for transformation of plants. Cloning in E. Coli, transfer to agrobacteria, integrates in the plant genome. Kan resistance in bacteria, confers Basta resistance in plants. Contains the pENTCUP2 promoter upstream of MCS for constitutive expression of gene of interest.<br>
+Vector for transformation of plants. Cloning in ''E. Coli'', transfer to agrobacteria, integrates in the plant genome. Kanamycin resistance in bacteria, confers Basta resistance in plants. Contains the pENTCUP2 promoter upstream of MCS for constitutive expression of gene of interest.<br>
 '''Confirmed functions''':
 * Kan resistance in bacteria: confirmed by transformation and colony growth of ''E. coli'' (DH5alpha) and ''Agrobacterium tumefaciens''
-* Basta resistance in plants: confirmed by selective germination of transformed Arabidopsis seeds on basta agar plates (See photos [http://2010.igem.org/Team:Harvard/results]).<br><br>
+* Basta resistance in plants: confirmed by selective germination of transformed ''Arabidopsis'' seeds on basta agar plates (See photos [http://2010.igem.org/Team:Harvard/results]).<br><br>
-Confirmation pending: Function of the ENTCUP2 promoter will be tested by checking for expression for several BioBrick parts in adult arabidopsis plants. These parts include...
+Confirmation pending: Function of the ENTCUP2 promoter will be tested by checking for expression for several BioBrick parts in adult ''Arabidopsis'' plants. These parts include...
 * miraculin ([[Part:BBa_K382021|BBa_K382021]])
 * brazzein ([[Part:BBa_K382052|BBa_K382052]])
@@ Line 16: / Line 16: @@
 <br>
-Successfully transfected basta resistant ''Arabidopsis'' plants, such as the one shown below (brazzein [[Part:BBa_K382052|BBa_K382052]]), will be tested for BioBrick expression once it has grown large enough for tissue sampling:
+Successfully transfected basta resistant ''Arabidopsis'' plants, such as the one shown below (brazzein [[Part:BBa_K382052|BBa_K382052]]), will be tested for BioBrick expression once they have grown large enough for tissue sampling:
 <br>