Difference between revisions of "Part:BBa K3040011"
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We create a hybrid Fatty acid/acyl-CoA-regulated promoter by the combination of pLac with a modified promoter pFadBA. It is pointed out by the previous iGEM team, UPF CRG Barcelona_2018 iGEM, that the native promoter pFadBA has great leakage and is a weak promoter that isn’t ideal enough for our system. Therefore, to enhance the sensitivity and reduce the leakage of the promoter pFadBA for making it more suitable for our system, we improved our promoter based on the previous pfadBA DNA sequence from NTU_Taida 2012.  
 
We create a hybrid Fatty acid/acyl-CoA-regulated promoter by the combination of pLac with a modified promoter pFadBA. It is pointed out by the previous iGEM team, UPF CRG Barcelona_2018 iGEM, that the native promoter pFadBA has great leakage and is a weak promoter that isn’t ideal enough for our system. Therefore, to enhance the sensitivity and reduce the leakage of the promoter pFadBA for making it more suitable for our system, we improved our promoter based on the previous pfadBA DNA sequence from NTU_Taida 2012.  
 
pFL1 promoter has a fadR recognition site between its -10 and -35 region, and a promoter pLac behind -10 region. pFL1 promoter is designed to respond to changes of both fatty acid and an exogenous inducer, IPTG. The hybrid promoter pFL1 is fully activated only when both the fatty acids and IPTG are present.
 
pFL1 promoter has a fadR recognition site between its -10 and -35 region, and a promoter pLac behind -10 region. pFL1 promoter is designed to respond to changes of both fatty acid and an exogenous inducer, IPTG. The hybrid promoter pFL1 is fully activated only when both the fatty acids and IPTG are present.
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3040013 SequenceAndFeatures</partinfo>
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===Functional Parameters===
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<partinfo>BBa_K3040013 parameters</partinfo>
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==Reference==
 
==Reference==
 
ZHANG, Fuzhong; CAROTHERS, James M.; KEASLING, Jay D. Design of a dynamic sensor-regulator system for production of chemicals and fuels derived from fatty acids. Nature biotechnology, 2012, 30.4: 354.
 
ZHANG, Fuzhong; CAROTHERS, James M.; KEASLING, Jay D. Design of a dynamic sensor-regulator system for production of chemicals and fuels derived from fatty acids. Nature biotechnology, 2012, 30.4: 354.

Revision as of 12:58, 19 October 2019

One FadR binding site with pLac promoter regulating downstream RFP

We create a hybrid Fatty acid/acyl-CoA-regulated promoter by the combination of pLac with a modified promoter pFadBA. It is pointed out by the previous iGEM team, UPF CRG Barcelona_2018 iGEM, that the native promoter pFadBA has great leakage and is a weak promoter that isn’t ideal enough for our system. Therefore, to enhance the sensitivity and reduce the leakage of the promoter pFadBA for making it more suitable for our system, we improved our promoter based on the previous pfadBA DNA sequence from NTU_Taida 2012. pFL1 promoter has a fadR recognition site between its -10 and -35 region, and a promoter pLac behind -10 region. pFL1 promoter is designed to respond to changes of both fatty acid and an exogenous inducer, IPTG. The hybrid promoter pFL1 is fully activated only when both the fatty acids and IPTG are present.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Unknown
  • 21
    INCOMPATIBLE WITH RFC[21]
    Unknown
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 755
    Illegal AgeI site found at 867
  • 1000
    COMPATIBLE WITH RFC[1000]



Reference

ZHANG, Fuzhong; CAROTHERS, James M.; KEASLING, Jay D. Design of a dynamic sensor-regulator system for production of chemicals and fuels derived from fatty acids. Nature biotechnology, 2012, 30.4: 354.