Difference between revisions of "Part:BBa K2092002:Design"

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===Design Notes===
 
===Design Notes===
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To add the missing XbaI and SpeI restriction sites, the following two primers were designed:
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XbaI and SpeI restriction sites were missing in the original BioBrick by 2011 iGEM DTU-Denmark.  To add the following sites, two primers were designed:
 +
 
 
alcA forward primer = 5'- CGG AAT TCG CGT CTA GAA CGT CGC TCT CCC CGA TGA C -3'  
 
alcA forward primer = 5'- CGG AAT TCG CGT CTA GAA CGT CGC TCT CCC CGA TGA C -3'  
 +
 
alcA reverse primer =  
 
alcA reverse primer =  
 
5'- AAC TGC AGA AAC CAA TGC ATT GGA CTA GTT TTT GAG GCG AGG TGA TAG GAT TGG -3'
 
5'- AAC TGC AGA AAC CAA TGC ATT GGA CTA GTT TTT GAG GCG AGG TGA TAG GAT TGG -3'
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===Source===
 
===Source===
  
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Aspergillus nidulans, sequence found through
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2011 iGEM DTU-Denmark 2
  
 
===References===
 
===References===
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[1] Panozzo, C., Capuano, V., Fillinger, S. and Felenbok, B. (1997). The zinc binuclear cluster Activator AlcR is able to bind to single sites but requires multiple repeated sites for synergistic activation of the alcA gene in Aspergillus nidulans. Journal of Biological Chemistry, 272(36), 22859–22865.
 
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[2] Kinkema, M., Geijskes, R.J., Shand, K., Coleman, H.D., De Lucca, P.C., Palupe, A., Harrison, M.D., Jepson, I., Dale, J.L. and Sainz, M.B. (2013). An improved chemically inducible gene switch that functions in the monocotyledonous plant sugar cane. Plant Molecular Biology, 84(4-5), 443–454.
 
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[3] Hemmati, H. and Basu, C. (2015). Transcriptional analyses of an ethanol inducible promoter in Escherichia coli and tobacco for production of cellulase and green fluorescent protein. Biotechnology & Biotechnological Equipment, 29(6), 1043–1052.
 

Latest revision as of 19:25, 29 October 2016


PalcA, improved alcR inducible promoter from A. nidulans


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 274
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

XbaI and SpeI restriction sites were missing in the original BioBrick by 2011 iGEM DTU-Denmark. To add the following sites, two primers were designed:

alcA forward primer = 5'- CGG AAT TCG CGT CTA GAA CGT CGC TCT CCC CGA TGA C -3'

alcA reverse primer = 5'- AAC TGC AGA AAC CAA TGC ATT GGA CTA GTT TTT GAG GCG AGG TGA TAG GAT TGG -3'

Source

2011 iGEM DTU-Denmark 2

References