Difference between revisions of "Part:BBa K1856000"
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<partinfo>BBa_K1856000 short</partinfo>
 
<partinfo>BBa_K1856000 short</partinfo>
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[https://static.igem.org/mediawiki/2015/8/82/Expression_Level_for_melA-citrine_%28pKT230-Lic%29.jpg]
 
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bacA promoter fused to citrine
 
  
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bacA promoter assembled to citrine and T7 terminator
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===Usage and Biology===
 
===Usage and Biology===
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bacA is an inducible promoter native to rhizobium species (notably R. tropici and S. meliloti) that is induced by flavonoids. This construct has been successfully cloned into E. coli using the broad-host range vector pKT230, a RSF1010 derived plasmid. Leaky expression of citrine was observed.
  
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The construct has also been cloned into S. meliloti.
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K1856000 SequenceAndFeatures</partinfo>
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==Description==
 
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Members of the Anderson promoter collection are suitable for general protein expression in ''E. coli'' and likely other prokaryotes.  The collection is known to cover a range of activities so by testing a few promoters it should be possible to find a promoter activity that suits your application.  The promoters were recovered from a library screen by [https://andersonlab.qb3.berkeley.edu/ Chris Anderson].
 
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Parts J23100 through J23119 are a family of constitutive promoter parts isolated from a small combinatorial library.  J23119 is the "consensus" promoter sequence and the strongest member of the family.  The NheI and AvrII restriction sites present within these promoter parts make them a scaffold for further modification. 
 
  
 
==Characterization==
 
==Characterization==
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[[Image:https://static.igem.org/mediawiki/2015/8/82/Expression_Level_for_melA-citrine_%28pKT230-Lic%29.jpg|300px|right]]
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[[Image:Expression_Level_for_melA-citrine_(pKT230-Lic).jpg|300px|right]]
 
===Measured strengths===
 
===Measured strengths===
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Reported activities of the promoters are given as the relative fluorescence of these plasmids in strain TG1 grown in LB media to saturation. See part [https://parts.igem.org/Part:BBa_J61002 J61002] for details on their use.
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Leaky expression of BacA was observed See part [https://parts.igem.org/Part:BBa_J61002 J61002] for details on their use.
  
 
==Obtaining the Anderson promoter collection==
 
==Obtaining the Anderson promoter collection==
 
[[Image:Expression_Level_for_melA-citrine_%28pKT230-Lic%29.jpg|right|150px|right]]
 
[[Image:Expression_Level_for_melA-citrine_%28pKT230-Lic%29.jpg|right|150px|right]]
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The sequences of the Anderson promoters can be found via the table below.  To obtain the physical DNA, we recommend two approaches - <br>
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The sequences of construct can be found via the table below.  The physical DNA can be obtained from:
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'''Via ''de novo'' synthesis''': Since the promoters are short sequences, they can be easily and cheaply ordered as two single-stranded complementary oligo's and annealed.  See [[Help:Promoters/Construction|here]] for a tutorial on how to construct short parts via oligo annealing.
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'''Via request''': The Yale iGEM team has the promoter cloned
  
 
'''Via the Registry distribution''': The promoters are included in the Registry distribution.  All parts except <partinfo>J23119</partinfo> are present in plasmid <partinfo>J61002</partinfo>.  This places the RFP downstream of the promoter.  Part <partinfo>J23119</partinfo> is present in <partinfo>pSB1A2</partinfo>.
 
'''Via the Registry distribution''': The promoters are included in the Registry distribution.  All parts except <partinfo>J23119</partinfo> are present in plasmid <partinfo>J61002</partinfo>.  This places the RFP downstream of the promoter.  Part <partinfo>J23119</partinfo> is present in <partinfo>pSB1A2</partinfo>.
 
<br style="clear:both" />
 
<br style="clear:both" />
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K1856000 SequenceAndFeatures</partinfo>

Revision as of 22:36, 20 September 2015

bacA-citrine-T7 terminator

bacA promoter assembled to citrine and T7 terminator

Usage and Biology

bacA is an inducible promoter native to rhizobium species (notably R. tropici and S. meliloti) that is induced by flavonoids. This construct has been successfully cloned into E. coli using the broad-host range vector pKT230, a RSF1010 derived plasmid. Leaky expression of citrine was observed.

The construct has also been cloned into S. meliloti.



Characterization

Expression Level for melA-citrine (pKT230-Lic).jpg

Measured strengths

Leaky expression of BacA was observed See part J61002 for details on their use.

Obtaining the Anderson promoter collection

Expression Level for melA-citrine (pKT230-Lic).jpg

The sequences of construct can be found via the table below. The physical DNA can be obtained from:

Via request: The Yale iGEM team has the promoter cloned

Via the Registry distribution: The promoters are included in the Registry distribution. All parts except BBa_J23119 are present in plasmid BBa_J61002. This places the RFP downstream of the promoter. Part BBa_J23119 is present in pSB1A2.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 920
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 920
    Illegal NotI site found at 945
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 920
    Illegal BamHI site found at 914
    Illegal XhoI site found at 954
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 920
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 920
  • 1000
    COMPATIBLE WITH RFC[1000]