Difference between revisions of "Part:BBa K1758310:Design"

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===Design Notes===
 
===Design Notes===
 
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We used the codon optimized, synthesized Sequence of chrB  from iGEM Team BIT 2013(<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K346002 " target="_blank"> BBa_K346002 </a>) under the control of a constitutive Primer(<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K346002" target="_blank"> BBa_K346002</a>). We designed the used primes, in specially split primers, for Gibson Assembly to ligate the constitutive Promoter and merR with pSB1C3. For this aim we designed four primers for the generation of homologous overlaps between the synthesized constitutive promoter+ merR and the pSB1C3:  
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We used the codon optimized, synthesized Sequence of chrB  from iGEM Team BIT 2013(<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K346001 " target="_blank"> BBa_K346001 </a>) under the control of a constitutive Primer(<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K346002" target="_blank"> BBa_K346002</a>). We designed the used primes, in specially split primers, for Gibson Assembly to ligate the constitutive Promoter and merR with pSB1C3. For this aim we designed four primers for the generation of homologous overlaps between the synthesized constitutive promoter+ merR and the pSB1C3:  
 
<p> <a href="http://2015.igem.org/Team:Bielefeld-CeBiTec/Primers#cm_fwd" target="_blank"> <a href="http://2015.igem.org/Team:Bielefeld-CeBiTec/Primers#cm_fwd" target="_blank">cm_fwd</a> (CGGCATCAGCACCTTGTC)</p>
 
<p> <a href="http://2015.igem.org/Team:Bielefeld-CeBiTec/Primers#cm_fwd" target="_blank"> <a href="http://2015.igem.org/Team:Bielefeld-CeBiTec/Primers#cm_fwd" target="_blank">cm_fwd</a> (CGGCATCAGCACCTTGTC)</p>
  

Revision as of 02:43, 19 September 2015

Chromium repressor under control of constitutive promoter and strong RBS


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 55
    Illegal NheI site found at 966
    Illegal NheI site found at 989
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 124
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We used the codon optimized, synthesized Sequence of chrB from iGEM Team BIT 2013( BBa_K346001 ) under the control of a constitutive Primer( BBa_K346002). We designed the used primes, in specially split primers, for Gibson Assembly to ligate the constitutive Promoter and merR with pSB1C3. For this aim we designed four primers for the generation of homologous overlaps between the synthesized constitutive promoter+ merR and the pSB1C3:

cm_fwd (CGGCATCAGCACCTTGTC)

pSB1C3_chrB_rev (CTTTTGAAAAAGGCACTCTGACCGTTTGACTCTAGAAGCGGCCGCGAAT)

cm_rev (TATACGCAAGGCGACAAG)

pSB1C3_kPrm_fwd (CTAGGACTGAGCTAGCTGTCAATACTAGTAGCGGCCGCTGCA)

Source

The original source of our chromate repressor is Ochrobactrum tritici 5bvl1. We used a codon optimized version which we got synthesised by IDT.

References