Difference between revisions of "Part:BBa K2374003:Design"

Revision as of 11:54, 30 October 2017

ple (Tyrosine 3-monooxygenase, TH) -> (fruit fly)

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 1157
Illegal XhoI site found at 289
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 535
Illegal BsaI site found at 1501
Illegal BsaI.rc site found at 253
Illegal BsaI.rc site found at 1019
Illegal BsaI.rc site found at 1168

Design Notes

According to our experiment results to judge, the ple coding sequence is hard to clone from Drosophila 's cDNA library because of its multi-segment repeats. So we recommend that you obtain from the constructed plasmid, or synthesize it directly. We ordered a synthetic ple from GENEWIZ®, and cloned it into pUAST vector with two restriction sites: EcoRI and XbaI.

We cloned UAS-TH into shipping backbone pSB1C3 with In-Fusion. Here shows the restriction endonuclease digestion image of pSB1C3-UAS-TH.

We also did one site direct mutagenesis for submission:
Pst I (647) CTGCAG->CTGCTG

Source

GeneCards®: The Human Gene Database [http://www.genecards.org/cgi-bin/carddisp.pl?gene=TH]

NCBI [1]

FlyBase [http://flybase.org/reports/FBgn0005626.html]

References

Janice A. Fischer, et al. GAL4 activates transcription in Drosophila. Nature 332, 853 - 856 (1988)

@@ Line 8: / Line 8: @@
 ===Design Notes===
+[[File:2017tongji_image_registry_TH_base.png|right|300px|标题]]
 According to our experiment results to judge, the '''ple''' coding sequence is hard to clone from ''Drosophila'' 's cDNA library because of its multi-segment repeats.
 So we recommend that you obtain from the constructed plasmid, or synthesize it directly.