Difference between revisions of "Part:BBa K3402053"
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This device is composed of up<i>SBLE</i>([https://parts.igem.org/Part:BBa_K3402030# BBa_K3402030]), Six site([https://parts.igem.org/Part:BBa_K3402032# BBa_K3402032]), P<i>galk</i>([https://parts.igem.org/Part:BBa_K3402033# BBa_K3402033]), <i>Rec</i>([https://parts.igem.org/Part:BBa_K3402034# BBa_K3402034]), T<i>gki</i>([https://parts.igem.org/Part:BBa_K3402019# BBa_K3402019]), <i>hph</i>([https://parts.igem.org/Part:BBa_K3402012# BBa_K3402012]), P<i>tef1</i>([https://parts.igem.org/Part:BBa_K3402007# BBa_K3402007]), <i>Cas9</i>([https://parts.igem.org/Part:BBa_K3402023# BBa_K3402023]), <i>NLS</i>([https://parts.igem.org/Part:BBa_K3402008# BBa_K3402008]), T<i>syn7</i>([https://parts.igem.org/Part:BBa_K3402001# BBa_K3402001]), do<i>SBLE</i>([https://parts.igem.org/Part:BBa_K3402031# BBa_K3402031]).
 
This device is composed of up<i>SBLE</i>([https://parts.igem.org/Part:BBa_K3402030# BBa_K3402030]), Six site([https://parts.igem.org/Part:BBa_K3402032# BBa_K3402032]), P<i>galk</i>([https://parts.igem.org/Part:BBa_K3402033# BBa_K3402033]), <i>Rec</i>([https://parts.igem.org/Part:BBa_K3402034# BBa_K3402034]), T<i>gki</i>([https://parts.igem.org/Part:BBa_K3402019# BBa_K3402019]), <i>hph</i>([https://parts.igem.org/Part:BBa_K3402012# BBa_K3402012]), P<i>tef1</i>([https://parts.igem.org/Part:BBa_K3402007# BBa_K3402007]), <i>Cas9</i>([https://parts.igem.org/Part:BBa_K3402023# BBa_K3402023]), <i>NLS</i>([https://parts.igem.org/Part:BBa_K3402008# BBa_K3402008]), T<i>syn7</i>([https://parts.igem.org/Part:BBa_K3402001# BBa_K3402001]), do<i>SBLE</i>([https://parts.igem.org/Part:BBa_K3402031# BBa_K3402031]).
  
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===Usage and Biology===
 
===Usage and Biology===

Revision as of 08:55, 21 October 2020


Self-excising hygromycin marker cassette

This device is composed of upSBLE(BBa_K3402030), Six site(BBa_K3402032), Pgalk(BBa_K3402033), Rec(BBa_K3402034), Tgki(BBa_K3402019), hph(BBa_K3402012), Ptef1(BBa_K3402007), Cas9(BBa_K3402023), NLS(BBa_K3402008), Tsyn7(BBa_K3402001), doSBLE(BBa_K3402031).

File:.png

Usage and Biology

We designed the self-excising hygromycin marker cassette which can be used to excise hygromycin resistance gene after screening. upSBLE and doSBLE are homologous arms which are used to construct device by homology directed repair. Pgalk is a galactose-induced promoter, which can control the expression of serine recombinant enzyme (Rec). According to the β-Rec/six self-excising system, serine recombinant enzyme will recognize two adjacent six sites in the same DNA strand and specifically cut off the fragments between two adjacent six sequences.
As a result, the resistance gene can be recycled and the engineered yeast will never show hygromycin resistance.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 858
    Illegal XbaI site found at 2596
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1380
    Illegal BamHI site found at 445
    Illegal BamHI site found at 3783
    Illegal XhoI site found at 2214
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 858
    Illegal XbaI site found at 2596
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 858
    Illegal XbaI site found at 2596
    Illegal NgoMIV site found at 523
  • 1000
    COMPATIBLE WITH RFC[1000]