Difference between revisions of "Part:BBa K2997003"
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The construction and digestion of DNA gel shows that the size of the pchR-GFP composite part was as expected.  
 
The construction and digestion of DNA gel shows that the size of the pchR-GFP composite part was as expected.  
  
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The pchR-GFP composite part (BBa_K2997008) was constructed from IDT DNA synthesis. After undergoing a few rounds of overlap PCR to combine 3 separate fragments into one long composite part, the part is then inserted into pSB1C3 and electroporated into E. coli Top 10 wild type. After allowing the transformed ''E. coli'' Top 10 to grow on LB+Cm plates, colony PCR and sequencing was carried out to determine the clone we got was positive or not. The colony PCR results indicated that the E. coli has pSB1C3 with pchR-GFP. However, the sequencing results indicated that random mutations have occurred on the pchR-GFP composite part.  
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The pchR-GFP composite part (BBa_K2997008) was constructed from IDT DNA synthesis. After undergoing a few rounds of overlap PCR to combine 3 separate fragments into one long composite part, the part is then inserted into pSB1C3 and electroporated into E. coli Top 10 wild type. After allowing the transformed <i>E. coli</i> Top 10 to grow on LB+Cm plates, colony PCR and sequencing was carried out to determine the clone we got was positive or not. The colony PCR results indicated that the E. coli has pSB1C3 with pchR-GFP. However, the sequencing results indicated that random mutations have occurred on the pchR-GFP composite part.  
  
  

Revision as of 12:39, 19 October 2019


Sigma 54-dependent transcriptional regulator (pchR)

The construction and digestion of DNA gel shows that the size of the pchR-GFP composite part was as expected.

The pchR-GFP composite part (BBa_K2997008) was constructed from IDT DNA synthesis. After undergoing a few rounds of overlap PCR to combine 3 separate fragments into one long composite part, the part is then inserted into pSB1C3 and electroporated into E. coli Top 10 wild type. After allowing the transformed E. coli Top 10 to grow on LB+Cm plates, colony PCR and sequencing was carried out to determine the clone we got was positive or not. The colony PCR results indicated that the E. coli has pSB1C3 with pchR-GFP. However, the sequencing results indicated that random mutations have occurred on the pchR-GFP composite part.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1738
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1151
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1196
    Illegal AgeI site found at 179
    Illegal AgeI site found at 1079
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1105