Difference between revisions of "Part:BBa K592008:Design"

(→‎Design Notes)
 
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===Design Notes===
 
===Design Notes===
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The most common DH5alpha and TOP10 used in assembly are not LacI-constitutive (LacIq) strains. Thus, any ORF placed downstream to this promoter will be massively transcribed, slowing bacterial growth. Therefore, it's recommended to employ low-copy number plasmids or even LacIq strains when adding genes to this promoter.
+
The most common DH5alpha and TOP10 used in assembly are not LacI-constitutive (LacIq) strains. Thus, any ORF placed downstream to this promoter will be massively transcribed, slowing bacterial growth. Therefore, it's recommended to employ low copy number plasmids or LacIq strains when adding genes to this promoter.
  
 
===Source===
 
===Source===
  
−
Phage T5
+
Sequence derived from the pJexpress401 plasmid from DNA 2.0. The sequence is ‘IP-free’.
  
 
===References===
 
===References===
 +
 +
[http://www.ncbi.nlm.nih.gov/pubmed/8045263] Oehler S, Amouyal M, Kolkhof P, von Wilcken-Bergmann B, Muller-Hill B (1994) Quality and position of the three lac operators of E. coli define efficiency of repression. EMBO J 13: 3348-3355.
 +
 +
[http://www.ncbi.nlm.nih.gov/pubmed/3900050] Gentz R, Bujard H (1985) Promoters Recognized by Escherichia-Coli Rna-Polymerase Selected by Function - Highly Efficient Promoters from Bacteriophage-T5. J Bacteriol 164: 70-77.
 +
 +
[http://www.ncbi.nlm.nih.gov/pubmed/3057497] Lanzer M, Bujard H (1988) Promoters Largely Determine the Efficiency of Repressor Action. P Natl Acad Sci USA 85: 8973-8977.
 +
 +
[https://www.dna20.com/pJexpress.php] List of DNA 2.0 Bacterial Expression Vectors, including pJexpress401.

Latest revision as of 13:47, 22 September 2011

T5-lac Promoter


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 most common DH5alpha and TOP10 used in assembly are not LacI-constitutive (LacIq) strains. Thus, any ORF placed downstream to this promoter will be massively transcribed, slowing bacterial growth. Therefore, it's recommended to employ low copy number plasmids or LacIq strains when adding genes to this promoter.

Source

Sequence derived from the pJexpress401 plasmid from DNA 2.0. The sequence is ‘IP-free’.

References

[http://www.ncbi.nlm.nih.gov/pubmed/8045263] Oehler S, Amouyal M, Kolkhof P, von Wilcken-Bergmann B, Muller-Hill B (1994) Quality and position of the three lac operators of E. coli define efficiency of repression. EMBO J 13: 3348-3355.

[http://www.ncbi.nlm.nih.gov/pubmed/3900050] Gentz R, Bujard H (1985) Promoters Recognized by Escherichia-Coli Rna-Polymerase Selected by Function - Highly Efficient Promoters from Bacteriophage-T5. J Bacteriol 164: 70-77.

[http://www.ncbi.nlm.nih.gov/pubmed/3057497] Lanzer M, Bujard H (1988) Promoters Largely Determine the Efficiency of Repressor Action. P Natl Acad Sci USA 85: 8973-8977.

[1] List of DNA 2.0 Bacterial Expression Vectors, including pJexpress401.