Part:BBa_K4325010
pDawn without LVA tag
Description
Our reduce the leakage level of pDawn system by removing the degradation tag LVA in the original repressor cI .
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 2171
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 63
Illegal NgoMIV site found at 195
Illegal NgoMIV site found at 289
Illegal NgoMIV site found at 582
Illegal NgoMIV site found at 1076
Illegal NgoMIV site found at 1094
Illegal NgoMIV site found at 1184
Illegal AgeI site found at 414
Illegal AgeI site found at 1542 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1643
Illegal BsaI.rc site found at 525
2022 SZPT-China
Biology
Our goal is to reduce the leakage level of pDawn system by removing the degradation tag LVA in the original repressor cI as increasing the concentration of cI could enhance the repression of PR promoter.
Usage
The plasmid containing the blue light induced kill switch was transformed into E. coli TOP10. Sixteen colonies were picked and grown on two new LB plates at 37 °C for 16 h. One plate was placed in the dark and the other was placed under blue light. The number of the candidates that survived the dark but failed to grow under blue light but survived in the dark was counted. For the E. Coli strain containing pDawn(cI-LVA)-RBS070-LKD-pSEVA331, the number is 5. For pDawn(cI)-RBS070-LKD-pSEVA331, the number is 10. The leakage expression of the lysis cassette LKD could be associated with this discrepancy in functionality. The new pDawn in which the LVA tag of cI is deleted improves the effectiveness of the blue light responsive lysis system.
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