Difference between revisions of "Part:BBa K3402052"

 
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<partinfo>BBa_K3402052 short</partinfo>
 
<partinfo>BBa_K3402052 short</partinfo>
  
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This device is composed of promoter Ptef1(BBa_K3402007), Cas9(BBa_K3402023)&#65292;NLS(BBa_K3402008), terminator Tsyn7(BBa_K3402001), HDV(BBa_K3402028), sgGFP(BBa_K3402024), HH and RS(BBa_K3402029), Ptef1(BBa_K3402007), upGFP(BBa_K3402025), hygromycin resistance gene hph(BBa_K3402012) and doGFP(BBa_K3402026).
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This device is composed of promoter Ptef1([https://parts.igem.org/Part:BBa_K3402007# BBa_K3402007]), Cas9([https://parts.igem.org/Part:BBa_K3402023# BBa_K3402023]), NLS([https://parts.igem.org/Part:BBa_K3402008# BBa_K3402008]), terminator Tsyn7([https://parts.igem.org/Part:BBa_K3402001# BBa_K3402001]), HDV([https://parts.igem.org/Part:BBa_K3402028# BBa_K3402028]), sgGFP([https://parts.igem.org/Part:BBa_K3402024# BBa_K3402024]), HH and RS([https://parts.igem.org/Part:BBa_K3402029# BBa_K3402029]), Ptef1([https://parts.igem.org/Part:BBa_K3402007# BBa_K3402007]), upGFP([https://parts.igem.org/Part:BBa_K3402025# BBa_K3402025]), hygromycin resistance gene hph([https://parts.igem.org/Part:BBa_K3402012# BBa_K3402012]) and doGFP([https://parts.igem.org/Part:BBa_K3402026# BBa_K3402026]).
  
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===Usage and Biology===
 
===Usage and Biology===
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After the construction of yeGFP expression device, we constructed a yeGFP editing device. We use Ptef1 as the promoter to express Cas9 protein and we add upGFP and doGFP to edit GFP site. The hygromycin resistance gene was built between the upGFP and the doGFP as the marker gene to determine if the transformation is successful.
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Then the transformed yeasts are coated on a plate containing hypopycin. After incubation, there are a lot of transformants on the plate. Then observe the green fluorescence of the transformants. None of them shows green fluorescence.
  
 
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Revision as of 12:29, 20 October 2020


GFP editing expression cassette

This device is composed of promoter Ptef1(BBa_K3402007), Cas9(BBa_K3402023), NLS(BBa_K3402008), terminator Tsyn7(BBa_K3402001), HDV(BBa_K3402028), sgGFP(BBa_K3402024), HH and RS(BBa_K3402029), Ptef1(BBa_K3402007), upGFP(BBa_K3402025), hygromycin resistance gene hph(BBa_K3402012) and doGFP(BBa_K3402026).

Usage and Biology

After the construction of yeGFP expression device, we constructed a yeGFP editing device. We use Ptef1 as the promoter to express Cas9 protein and we add upGFP and doGFP to edit GFP site. The hygromycin resistance gene was built between the upGFP and the doGFP as the marker gene to determine if the transformation is successful.
Then the transformed yeasts are coated on a plate containing hypopycin. After incubation, there are a lot of transformants on the plate. Then observe the green fluorescence of the transformants. None of them shows green fluorescence.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 5609
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1736
    Illegal BglII site found at 5300
    Illegal XhoI site found at 1051
    Illegal XhoI site found at 1786
    Illegal XhoI site found at 2149
    Illegal XhoI site found at 5101
    Illegal XhoI site found at 6701
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 5609
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 5609
    Illegal NgoMIV site found at 2563
    Illegal NgoMIV site found at 3667
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1507
    Illegal BsaI site found at 2224
    Illegal SapI.rc site found at 7265