Difference between revisions of "Part:BBa K2971010"
 
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<partinfo>BBa_K2971010 short</partinfo>
 
<partinfo>BBa_K2971010 short</partinfo>
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crtEB
 
  
 
This part contains the sequences of crtE and crtB connected by a short linker sequence. The stop codon
 
This part contains the sequences of crtE and crtB connected by a short linker sequence. The stop codon
 
of each gene has been removed to express the genes in a single open reading frame. The enzymes
 
of each gene has been removed to express the genes in a single open reading frame. The enzymes
 
encoded by the genes supply the precursor of carotenoids when expressed in Escherichia coli.
 
encoded by the genes supply the precursor of carotenoids when expressed in Escherichia coli.
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The composite part was made by fusing crtEB (BBa_K2971010) and crtI (BBa_K2971001) via Gibson cloning. The part was then transformed into competent cells of <i>E. coli</i>. The presence of the insert was screened with colony pcr (figure 2). The colony with the insert was analyzed by sequencing to check for potential mutations or frame-shifts.   
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<html>
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<img style="width:40% !important;" src="https://2019.igem.org/wiki/images/9/9d/T--UiOslo_Norway--EBcolonyPCR.jpg">
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<p>
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<strong>Figure 2: Colony PCR of cells transformed with crtEB</strong></br>
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Only one colony (1) showed the presence of the correct insert. 1 was then further investigated.
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</p>
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</html>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 02:07, 22 October 2019


crtE and crtB in a single open reading frame

This part contains the sequences of crtE and crtB connected by a short linker sequence. The stop codon of each gene has been removed to express the genes in a single open reading frame. The enzymes encoded by the genes supply the precursor of carotenoids when expressed in Escherichia coli.

The composite part was made by fusing crtEB (BBa_K2971010) and crtI (BBa_K2971001) via Gibson cloning. The part was then transformed into competent cells of E. coli. The presence of the insert was screened with colony pcr (figure 2). The colony with the insert was analyzed by sequencing to check for potential mutations or frame-shifts.

Figure 2: Colony PCR of cells transformed with crtEB
Only one colony (1) showed the presence of the correct insert. 1 was then further investigated.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1602
    Illegal BamHI site found at 841
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 43