Difference between revisions of "Part:BBa K1465201:Design"

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===Design Notes===
 
===Design Notes===
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Gibson-Assembly
 
  
 +
*Amplification of the pSB1C3-Backbone (<bbpart>BBa_J04450</bbpart>) using the specific Gibson-Primer [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#pSB1C3_pre_prkA pSB1C3_pre_prkA] and [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#pSB1C3_suf_prkA pSB1C3_suf_prkA]:<br>
  
−
Primer <a href="http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#Deletion_alanine_racemase" target="_blank">pSB1C3_pre_prkA</a> and <a href="http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#Deletion_alanine_racemase" target="_blank">pSB1C3_suf_prkA</a>
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'''pSB1C3_pre_prkA (43 bp)'''<br>
 +
5'-ACACGATCTGGCTTGCTCATCTCTAGAAGCGGCCGCGAATTCC-3'<br>
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'''pSB1C3_suf_prkA (43 bp)'''<br>
 +
5'-CGGTTGCCGCTAGCGTTTAATACTAGTAGCGGCCGCTGCAGTC-3'<br>
 +
 
 +
*[http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Gibson Gibson-Assembly] of the pSB1C3-Backbone and the gBlock containing the optimized coding sequence of the Phosphoribulokinase (''prkA'') from ''Synechococcus elangatus''.<br>
  
 
===Source===
 
===Source===
  
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Gensynthesis (gBlocks)
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Gensynthesis ([http://eu.idtdna.com/pages/products/genes/gblocks-gene-fragments gBlocks]) of the codon optimized sequence for ''Escherchia coli'' by using the [http://www.jcat.de/ Codon Adaptation Tool] and the [http://eu.idtdna.com/CodonOpt Codon Optimization from IDT]:
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 +
 
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BILD
  
 
===References===
 
===References===

Latest revision as of 17:38, 3 October 2014


Phosphoribulokinase prkA from Synechococcus elongatus


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1009
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 198
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 657
    Illegal AgeI site found at 928
    Illegal AgeI site found at 999
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

  • Amplification of the pSB1C3-Backbone (BBa_J04450) using the specific Gibson-Primer [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#pSB1C3_pre_prkA pSB1C3_pre_prkA] and [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#pSB1C3_suf_prkA pSB1C3_suf_prkA]:
pSB1C3_pre_prkA (43 bp)

5'-ACACGATCTGGCTTGCTCATCTCTAGAAGCGGCCGCGAATTCC-3'

pSB1C3_suf_prkA (43 bp)

5'-CGGTTGCCGCTAGCGTTTAATACTAGTAGCGGCCGCTGCAGTC-3'
  • [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Gibson Gibson-Assembly] of the pSB1C3-Backbone and the gBlock containing the optimized coding sequence of the Phosphoribulokinase (prkA) from Synechococcus elangatus.

Source

Gensynthesis ([http://eu.idtdna.com/pages/products/genes/gblocks-gene-fragments gBlocks]) of the codon optimized sequence for Escherchia coli by using the [http://www.jcat.de/ Codon Adaptation Tool] and the [http://eu.idtdna.com/CodonOpt Codon Optimization from IDT]:


BILD

References