Difference between revisions of "Part:BBa K2374007:Design"

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===Design Notes===
 
===Design Notes===
−
At the forward of the ple sequence, there is a ~150bp region which contains up to 5 copies of some specific sequences.
+
The mRNA of this gene must be cleaved to be translated into mature protein. At the forward of the ple coding sequence, there is a ~150bp region which contains up to 5 copies of some specific sequences.
 
This makes it difficult to design specific primers to clone this gene.
 
This makes it difficult to design specific primers to clone this gene.
 
We have tried many different sets of primers and different DNA Polymerases to clone ple from Drosophila's cDNA library.
 
We have tried many different sets of primers and different DNA Polymerases to clone ple from Drosophila's cDNA library.
 
Unfortunately we all failed to do this.
 
Unfortunately we all failed to do this.
 
There had been appeared some negative fragments which even had the same size of the real one.
 
There had been appeared some negative fragments which even had the same size of the real one.
 +
Finally we ordered the synthetic ple from GENEWIZ®
  
 
===Source===
 
===Source===
  
−
DROSOPHILA
+
NCBI: >NM_057550.4:453-2192 Drosophila melanogaster pale (ple), transcript variant B, mRNA
 +
 
 +
Synthetic
  
 
===References===
 
===References===
 +
 +
Hoskins,R.A.,at al. Sequence finishing and mapping of Drosophila melanogaster heterochromatin. Science 316 (5831), 1625-1628 (2007)

Latest revision as of 09:15, 29 October 2017


UAS-TH -> (fruit fly)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 13
    Illegal PstI site found at 256
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 13
    Illegal PstI site found at 256
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1540
    Illegal XhoI site found at 672
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 13
    Illegal PstI site found at 256
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 13
    Illegal PstI site found at 256
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 918
    Illegal BsaI site found at 1884
    Illegal BsaI.rc site found at 636
    Illegal BsaI.rc site found at 1402
    Illegal BsaI.rc site found at 1551


Design Notes

The mRNA of this gene must be cleaved to be translated into mature protein. At the forward of the ple coding sequence, there is a ~150bp region which contains up to 5 copies of some specific sequences. This makes it difficult to design specific primers to clone this gene. We have tried many different sets of primers and different DNA Polymerases to clone ple from Drosophila's cDNA library. Unfortunately we all failed to do this. There had been appeared some negative fragments which even had the same size of the real one. Finally we ordered the synthetic ple from GENEWIZ®

Source

NCBI: >NM_057550.4:453-2192 Drosophila melanogaster pale (ple), transcript variant B, mRNA

Synthetic

References

Hoskins,R.A.,at al. Sequence finishing and mapping of Drosophila melanogaster heterochromatin. Science 316 (5831), 1625-1628 (2007)