Difference between revisions of "Part:BBa K3970026"
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| + | <html><p style="font-size: 18px;word-wrap:break-word;">ATGACTAAATCTCATTCAGAAGAAGTGATTGTACCTGAGTTCAATTCTAGCGCAAAGGAATTACCAAGACCATTGGCCGAAAAGTGCCCGAGCATAATTAAGAAATTTATAAGCGCTTATGATGCTAAACCGGATTTTGTTGCTAGATCGCCTGGTAGAGTCAATCTAATTGGTGAACATATTGATTATTGTGACTTCTCGGTTTTACCTTTAGCTATTGATTTTGATATGCTTTGCGCCGTCAAAGTTTTGAACGAGAAAAATCCATCCATTACCTTAATAAATGCTGATCCCAAATTTGCTCAAAGGAAGTTCGATTTGCCGTTGGACGGTTCTTATGTCACAATTGATCCTTCTGTGTCGGACTGGTCTAATTACTTTAAATGTGGTCTCCATGTTGCTCACTCTTTTCTAAAGAAACTTGCACCGGAAAGGTTTGCCAGTGCTCCTCTGGCCGGGCTGCAAGTCTTCTGTGAGGGTGATGTACCAACTGGCAGTGGATTGTCTTCTTCGGCCGCATTCATTTGTGCCGTTGCTTTAGCTGTTGTTAAAGCGAATATGGGCCCTGGTTATCATATGTCCAAGCAAAATTTAATGCGTATTACGGTCGTTGCAGAACATTATGTTGGTGTTAACAATGGCGGTATGGATCAGGCTGCCTCTGTTTGCGGTGAGGAAGATCATGCTCTATACGTTGAGTTCAAACCGCAGTTGAAGGCTACTCCGTTTAAATTTCCGCAATTAAAAAACCATGAAATTAGCTTTGTTATTGCGAACACCCTTGTTGTATCTAACAAGTTTGAAACCGCCCCAACCAACTATAATTTAAGAGTGGTAGAAGTCACTACAGCTGCAAATGTTTTAGCTGCCACGTACGGTGTTGTTTTACTTTCTGGAAAAGAAGGATCGAGCACGAATAAAGGTAATCTAAGAGATTTCATGAACGTTTATTATGCCAGATATCACAACATTTCCACACCCTGGAACGGCGATATTGAATCCGGCATCGAACGGTTAACAAAGATGCTAGTACTAGTTGAAGAGTCTCTCGCCAATAAGAAACAGGGCTTTAGTGTTGACGATGTCGCACAATCCTTGAATTGTTCTCGCGAAGAATTCACAAGAGACTACTTAACAACATCTCCAGTGAGATTTCAAGTCTTAAAGCTATATCAGAGGGCTAAGCATGTGTATTCTGAATCTTTAAGAGTCTTGAAGGCTGTGAAATTAATGACTACAGCGAGCTTTACTGCCGACGAAGACTTTTTCAAGCAATTTGGTGCCTTGATGAACGAGTCTCAAGCTTCTTGCGATAAACTTTACGAATGTTCTTGTCCAGAGATTGACAAAATTTGTTCCATTGCTTTGTCAAATGGATCATATGGTTCCCGTTTGACCGGAGCTGGCTGGGGTGGTTGTACTGTTCACTTGGTTCCAGGGGGCCCAAATGGCAACATAGAAAAGGTAAAAGAAGCCCTTGCCAATGAGTTCTACAAGGTCAAGTACCCTAAGATCACTGATGCTGAGCTAGAAAATGCTATCATCGTCTCTAAACCAGCATTGGGCAGCTGTCTATATGAATTATAAATGAAAGCGTTAACGGCCAGGCAACAAGAGGTGTTTGATCTCATCCGTGATCACATCAGCCAGACAGGTATGCCGCCGACGCGTGCGGAAATCGCGCAGCGTTTGGGGTTCCGTTCCCCAAACGCGGCTGAAGAACATCTGAAGGCGCTGGCACGCAAAGGCGTTATTGAAATTGTTTCCGGCGCATCACGCGGGATTCGTCTGTTGCAGGAAGAGGAAGAAGGGTTGCCGCTGGTAGGTCGTGTGGCTGCCGGTGAACCACTTCTGGCGCAACAGCATATTGAAGGTCATTATCAGGTCGATCCTTCCTTATTCAAGCCGAATGCTGATTTCCTGCTGCGCGTCAGCGGGATGTCGATGAAAGATATCGGCATTATGGATGGTGACTTGCTGGCAGTGCATAAAACTCAGGATGTACGTAACGGTCAGGTCGTTGTCGCACGTATTGATGACGAAGTTACCGTTAAGCGCCTGAAAAAACAGGGCAATAAAGTCGAACTGTTGCCAGAAAATAGCGAGTTTAAACCAATTGTCGTTGACCTTCGTCAGCAGAGCTTCACCATTGAAGGGCTGGCGGTTGGGGTTATTCGCAACGGCGACTGGCTGTAAATGGAGTGTAATTTAGGAACCGAACATCCTAGTACAGATACGTGGAATCGTAGTAAAACGGAACAAGCGGTTGTGGACGCATTTGATGAATCGTTGTTTGGTGATGTAGCATCGGATATTGGATTTGAAACGTCGTTATATTCACATGCAGTTAAAACTGCTCCGTCTCCGCCTTGGGTAGCTAGCCCTAAAATTTTATATCAACAGTTAATACGGGATCTTGATTTTTCAGAAGGGCCGCGTTTACTATCATGTCTTGAAACCTGGAACGAGGATTTATTCTCATGTTTTCCTATTAATGAGGACCTATATTCCGATATGATGGTTTTATCCCCGGATCCAGATGACGTTATCTCAACCGTTTCAACCAAAGACCATGTTGAAATGTTTAATTTAACAACCCGGGGTTCCGTTCGATTGCCTAGTCCACCAAAGCAACCGACGGGGCTTCCAGCTTACGTTCAGGAGGTCCAGGATTCGTTTACCGTAGAACTACGCGCCCGGGAAGAAGCATACACAAAACTACTAGTTACTTATTGTAAATCGATTATACGTTATCTCCAAGGAACGGCGAAAAGGACGACAATAGGTCTTAATATACAAAACCCTGACCAGAAAGCTTACACGCAACTCAGGCAAAGTATTCTACTTAGATATTATCGTGAGGTGGCAAGTTTGGCGCGTCTTCTGTACCTACATTTATATTTAACCGTAACGCGTGAATTTTCCTGGCGTTTGTACGCCAGTCAATCTGCACACCCGGACGTGTTTGCGGCTTTAAAATTCACCTGGACCGAACGTCGACAGTTCACGTGTGCGTTTCATCCTGTATTATGCAACCACGGCATTGTGTTATTAGAAGGGAAACCACTAACAGCGTCTGCCTTGAGGGAAATAAATTACCGCCGCCGAGAACTGGGACTGCCTCTAGTTAGATGTGGTCTTGTTGAAGAAAACAAATCTCCGTTGGTTCAACAACCCTCATTTTCGGTTCATTTACCACGGTCGGTGGGTTTTCTTACCCACCACATTAAGCGTAAGTTAGACGCATATGCGGTCAAACATCCTCAAGAACCGAGACATGTACGAGCGGATCATCCTTACGCAAAAGTTGTTGAAAATAGAAACTACGGTAGTAGCATCGAAGCTATGATTTTAGCACCTCCGTCCCCATCCGAGATCCTGCCGGGGGACCCACCACGCCCACCCACGTGTGGGTTTTTAACGCGTTAAACGTCATTGGGGTAGAGGGTGTAAATAAAgcgaatttcttatgatttatgatttttattattaaataagttataaaaaaaataagtgtatacaaattttaaagtgactcttaggttttaaaacgaaaattcttattcttgagtaactctttcctgtaggtcaggttgctttctcaggtatagcatgaggtcgctcttattgaccacacctctaccgg</span> | ||
<partinfo>BBa_K3970026 SequenceAndFeatures</partinfo> | <partinfo>BBa_K3970026 SequenceAndFeatures</partinfo> | ||
Revision as of 01:58, 7 October 2021
pGAL1-LexA-VP16-tADH1
1.Usage and Biology
This part is the regulator of the following gene( cpcA, cpcE, cpcF). We learned that GAL1 is a commonly used inducible promoter in S. cerevisiae. The transcription of GAL genes involved in galactose metabolism in S. cerevisiae is tightly regulated, and they are not expressed in the absence of galactose. In the presence of galactose, the expression level of galactose was increased by about 1 ,000 fold. A large number of biochemical genetic analyses have preliminarily established the basic regulatory model of GAL gene expression. In addition, the expression of the GAL gene is also regulated by metabolite repression, thus forming a complex regulatory network system.LexA is a transcriptional regulator in yeast and VP16 is a transcription-activating enzyme that can enhance the expression of downstream genes.
We selected lexa-LEXO as a pair of inductor, and started transcription with GAL1 as the inducer promoter, and added a VP16 sequence after LexA to increase its expression.
2.Construction
ATGACTAAATCTCATTCAGAAGAAGTGATTGTACCTGAGTTCAATTCTAGCGCAAAGGAATTACCAAGACCATTGGCCGAAAAGTGCCCGAGCATAATTAAGAAATTTATAAGCGCTTATGATGCTAAACCGGATTTTGTTGCTAGATCGCCTGGTAGAGTCAATCTAATTGGTGAACATATTGATTATTGTGACTTCTCGGTTTTACCTTTAGCTATTGATTTTGATATGCTTTGCGCCGTCAAAGTTTTGAACGAGAAAAATCCATCCATTACCTTAATAAATGCTGATCCCAAATTTGCTCAAAGGAAGTTCGATTTGCCGTTGGACGGTTCTTATGTCACAATTGATCCTTCTGTGTCGGACTGGTCTAATTACTTTAAATGTGGTCTCCATGTTGCTCACTCTTTTCTAAAGAAACTTGCACCGGAAAGGTTTGCCAGTGCTCCTCTGGCCGGGCTGCAAGTCTTCTGTGAGGGTGATGTACCAACTGGCAGTGGATTGTCTTCTTCGGCCGCATTCATTTGTGCCGTTGCTTTAGCTGTTGTTAAAGCGAATATGGGCCCTGGTTATCATATGTCCAAGCAAAATTTAATGCGTATTACGGTCGTTGCAGAACATTATGTTGGTGTTAACAATGGCGGTATGGATCAGGCTGCCTCTGTTTGCGGTGAGGAAGATCATGCTCTATACGTTGAGTTCAAACCGCAGTTGAAGGCTACTCCGTTTAAATTTCCGCAATTAAAAAACCATGAAATTAGCTTTGTTATTGCGAACACCCTTGTTGTATCTAACAAGTTTGAAACCGCCCCAACCAACTATAATTTAAGAGTGGTAGAAGTCACTACAGCTGCAAATGTTTTAGCTGCCACGTACGGTGTTGTTTTACTTTCTGGAAAAGAAGGATCGAGCACGAATAAAGGTAATCTAAGAGATTTCATGAACGTTTATTATGCCAGATATCACAACATTTCCACACCCTGGAACGGCGATATTGAATCCGGCATCGAACGGTTAACAAAGATGCTAGTACTAGTTGAAGAGTCTCTCGCCAATAAGAAACAGGGCTTTAGTGTTGACGATGTCGCACAATCCTTGAATTGTTCTCGCGAAGAATTCACAAGAGACTACTTAACAACATCTCCAGTGAGATTTCAAGTCTTAAAGCTATATCAGAGGGCTAAGCATGTGTATTCTGAATCTTTAAGAGTCTTGAAGGCTGTGAAATTAATGACTACAGCGAGCTTTACTGCCGACGAAGACTTTTTCAAGCAATTTGGTGCCTTGATGAACGAGTCTCAAGCTTCTTGCGATAAACTTTACGAATGTTCTTGTCCAGAGATTGACAAAATTTGTTCCATTGCTTTGTCAAATGGATCATATGGTTCCCGTTTGACCGGAGCTGGCTGGGGTGGTTGTACTGTTCACTTGGTTCCAGGGGGCCCAAATGGCAACATAGAAAAGGTAAAAGAAGCCCTTGCCAATGAGTTCTACAAGGTCAAGTACCCTAAGATCACTGATGCTGAGCTAGAAAATGCTATCATCGTCTCTAAACCAGCATTGGGCAGCTGTCTATATGAATTATAAATGAAAGCGTTAACGGCCAGGCAACAAGAGGTGTTTGATCTCATCCGTGATCACATCAGCCAGACAGGTATGCCGCCGACGCGTGCGGAAATCGCGCAGCGTTTGGGGTTCCGTTCCCCAAACGCGGCTGAAGAACATCTGAAGGCGCTGGCACGCAAAGGCGTTATTGAAATTGTTTCCGGCGCATCACGCGGGATTCGTCTGTTGCAGGAAGAGGAAGAAGGGTTGCCGCTGGTAGGTCGTGTGGCTGCCGGTGAACCACTTCTGGCGCAACAGCATATTGAAGGTCATTATCAGGTCGATCCTTCCTTATTCAAGCCGAATGCTGATTTCCTGCTGCGCGTCAGCGGGATGTCGATGAAAGATATCGGCATTATGGATGGTGACTTGCTGGCAGTGCATAAAACTCAGGATGTACGTAACGGTCAGGTCGTTGTCGCACGTATTGATGACGAAGTTACCGTTAAGCGCCTGAAAAAACAGGGCAATAAAGTCGAACTGTTGCCAGAAAATAGCGAGTTTAAACCAATTGTCGTTGACCTTCGTCAGCAGAGCTTCACCATTGAAGGGCTGGCGGTTGGGGTTATTCGCAACGGCGACTGGCTGTAAATGGAGTGTAATTTAGGAACCGAACATCCTAGTACAGATACGTGGAATCGTAGTAAAACGGAACAAGCGGTTGTGGACGCATTTGATGAATCGTTGTTTGGTGATGTAGCATCGGATATTGGATTTGAAACGTCGTTATATTCACATGCAGTTAAAACTGCTCCGTCTCCGCCTTGGGTAGCTAGCCCTAAAATTTTATATCAACAGTTAATACGGGATCTTGATTTTTCAGAAGGGCCGCGTTTACTATCATGTCTTGAAACCTGGAACGAGGATTTATTCTCATGTTTTCCTATTAATGAGGACCTATATTCCGATATGATGGTTTTATCCCCGGATCCAGATGACGTTATCTCAACCGTTTCAACCAAAGACCATGTTGAAATGTTTAATTTAACAACCCGGGGTTCCGTTCGATTGCCTAGTCCACCAAAGCAACCGACGGGGCTTCCAGCTTACGTTCAGGAGGTCCAGGATTCGTTTACCGTAGAACTACGCGCCCGGGAAGAAGCATACACAAAACTACTAGTTACTTATTGTAAATCGATTATACGTTATCTCCAAGGAACGGCGAAAAGGACGACAATAGGTCTTAATATACAAAACCCTGACCAGAAAGCTTACACGCAACTCAGGCAAAGTATTCTACTTAGATATTATCGTGAGGTGGCAAGTTTGGCGCGTCTTCTGTACCTACATTTATATTTAACCGTAACGCGTGAATTTTCCTGGCGTTTGTACGCCAGTCAATCTGCACACCCGGACGTGTTTGCGGCTTTAAAATTCACCTGGACCGAACGTCGACAGTTCACGTGTGCGTTTCATCCTGTATTATGCAACCACGGCATTGTGTTATTAGAAGGGAAACCACTAACAGCGTCTGCCTTGAGGGAAATAAATTACCGCCGCCGAGAACTGGGACTGCCTCTAGTTAGATGTGGTCTTGTTGAAGAAAACAAATCTCCGTTGGTTCAACAACCCTCATTTTCGGTTCATTTACCACGGTCGGTGGGTTTTCTTACCCACCACATTAAGCGTAAGTTAGACGCATATGCGGTCAAACATCCTCAAGAACCGAGACATGTACGAGCGGATCATCCTTACGCAAAAGTTGTTGAAAATAGAAACTACGGTAGTAGCATCGAAGCTATGATTTTAGCACCTCCGTCCCCATCCGAGATCCTGCCGGGGGACCCACCACGCCCACCCACGTGTGGGTTTTTAACGCGTTAAACGTCATTGGGGTAGAGGGTGTAAATAAAgcgaatttcttatgatttatgatttttattattaaataagttataaaaaaaataagtgtatacaaattttaaagtgactcttaggttttaaaacgaaaattcttattcttgagtaactctttcctgtaggtcaggttgctttctcaggtatagcatgaggtcgctcttattgaccacacctctaccgg