Difference between revisions of "Part:BBa K3634018:Design"

 
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===Design Notes===
 
===Design Notes===
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Unusually, pre-existing parts on the registry for the ccdA antitoxin had differing sequences. Thus a new twin part was categorised with a different sequence.
 
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The wt sequence for the lac operon regulatory region was taken from Kalnins A. (1993) [GenBank accession no. J01636]. Within the CAP binding site, bases -66 (G) and -55 (C) of the wt binding region were substituted with A and T respectively to prevent binding of the CAP protein at low glucose concentrations. The wt -10 promoter sequence was also mutated from TATGTT to TATAAT in order to allow σ factor (RpoD) to bind without relying on further activation by the CAP protein (Reznikoff et al., 1978).
  
 
===Source===
 
===Source===
  
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The ccdA antitoxin is found natively in E.coli with sequence obtained from the pCoo plasmid (NCBI Accession Number: NC_007635.1). This was confirmed with sequence from NCBI Accession Number: NC_019000.  
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The lac operon is found natively in E.coli. The part sequence was taken from BBa_K1695000 who generated the part sequence through data obtained from Reznikoff et al. (1978) and Hirschel et al. (1980).
  
 
===References===
 
===References===
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 +
Jacob F., Monod J. 1961. Genetic Regulatory Mechanisms in the Synthesis of Proteins. J. Mol. Biol. 3: p818-356.
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 +
Reznikoff W. S., Abelson J. N. 1978. The lac promoter. The operon. 7: p221-243. DOI: 10.1101/0.221-243
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 +
Hirschel B.J., Shen V., Schlessinger D. 1980. Lactose Operon Transcription from Wild-Type and L8-UV5 lac Promoters in Escherichia coli Treated with Chloramphenicol. J. Bacteriol. 143(3): p1534-1537.
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City University of Hong Kong iGEM 2015 - https://parts.igem.org/Part:BBa_K1695000

Latest revision as of 11:41, 9 August 2020


PL8-UV5


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The wt sequence for the lac operon regulatory region was taken from Kalnins A. (1993) [GenBank accession no. J01636]. Within the CAP binding site, bases -66 (G) and -55 (C) of the wt binding region were substituted with A and T respectively to prevent binding of the CAP protein at low glucose concentrations. The wt -10 promoter sequence was also mutated from TATGTT to TATAAT in order to allow σ factor (RpoD) to bind without relying on further activation by the CAP protein (Reznikoff et al., 1978).

Source

The lac operon is found natively in E.coli. The part sequence was taken from BBa_K1695000 who generated the part sequence through data obtained from Reznikoff et al. (1978) and Hirschel et al. (1980).

References

Jacob F., Monod J. 1961. Genetic Regulatory Mechanisms in the Synthesis of Proteins. J. Mol. Biol. 3: p818-356.

Reznikoff W. S., Abelson J. N. 1978. The lac promoter. The operon. 7: p221-243. DOI: 10.1101/0.221-243

Hirschel B.J., Shen V., Schlessinger D. 1980. Lactose Operon Transcription from Wild-Type and L8-UV5 lac Promoters in Escherichia coli Treated with Chloramphenicol. J. Bacteriol. 143(3): p1534-1537.

City University of Hong Kong iGEM 2015 - https://parts.igem.org/Part:BBa_K1695000