Difference between revisions of "Part:BBa K1603004:Design"

 
(→‎Design Notes)
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===Design Notes===
 
===Design Notes===
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Codon optimized for saccharomyces cerevisiae and fused with a nuclear localization signal (NLS) for in vivo DNA repair. Also fused with a His-tag, allowing nickel based purification prior in vitro DNA repair.
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Synthetic gene ordered from IDT.  
  
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Codon optimized for ''Saccharomyces cerevisiae'' and fused with a nuclear localization signal (NLS) for in vivo DNA repair. Also fused with a His-tag, allowing nickel based purification prior in vitro DNA repair
  
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Primers for amplification with BioBrick Prefix in the FW primer and Suffix in the RV primer:
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FW:GCTTCTAGATGTCTCATAGACATCACCATCAC
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RV:AGCCTGCAGCGGCCGCTACTAGTATTAAAAGGGTAAATCATCTTCTTCTG
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The primers were designed to remove the EcoRI site from the insert when constructing the biobrick to reduce the amount of base pairs in the primers. The primers also add 3 extra protective basepairs to the PstI site in the insert.
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The construction of the final biobrick was initiated by cutting [https://parts.igem.org/Part:BBa_J04450 BBa_J04450] with XbaI, PstI, KnpI and FastAP and the insert with XbaI and PstI.
  
 
===Source===
 
===Source===

Revision as of 13:44, 18 September 2015


SSB - optimized for yeast


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 248
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 248
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 111
    Illegal XhoI site found at 875
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 248
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 248
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 482


Design Notes

Synthetic gene ordered from IDT.

Codon optimized for Saccharomyces cerevisiae and fused with a nuclear localization signal (NLS) for in vivo DNA repair. Also fused with a His-tag, allowing nickel based purification prior in vitro DNA repair


Primers for amplification with BioBrick Prefix in the FW primer and Suffix in the RV primer:

FW:GCTTCTAGATGTCTCATAGACATCACCATCAC

RV:AGCCTGCAGCGGCCGCTACTAGTATTAAAAGGGTAAATCATCTTCTTCTG


The primers were designed to remove the EcoRI site from the insert when constructing the biobrick to reduce the amount of base pairs in the primers. The primers also add 3 extra protective basepairs to the PstI site in the insert.


The construction of the final biobrick was initiated by cutting BBa_J04450 with XbaI, PstI, KnpI and FastAP and the insert with XbaI and PstI.

Source

Sequence adapted from Deinococcus Radiodurans. Ordered as synthetic gene.

References