Composite

Part:BBa_K3171172

Designed by: Yashika Venkatesh   Group: iGEM19_Groningen   (2019-10-16)
Revision as of 10:30, 21 October 2019 by Yashikavenkatesh (Talk | contribs)


Vibrio natriegens Native P1 promotor with mCherry

Vibrio natriegens has been reported to have a remarkable doubling time of 9.8 min. As is the case for the better-studied but slower-growing E. coli, V. natriegens increases its number of ribosomes with the growth rate in order to achieve its extraordinarily high rate of protein synthesis. Multiple mechanisms contribute to this high ribosome synthesis efficiency, including high rRNA gene copy number, strong promoters that contain near-consensus −10, −35, and UP elements and activation by the transcription factor Fis. In addition, V. natriegens rRNA promoters exhibit the relatively short-lived open-complex characteristic of rRNA promoters in E. coli, potentially contributing to the regulation of these promoters in vivo. The native promoter P1 is a constitutive promoter. We fused the the native P1 promoter with mCherry fluorescent protein with the help of the 3A assembly cloning method.

The P1 promoter with Biobrick prefix and suffix was cloned with mCherry to obtain a composite part that constitutively expresses mCherry Part:BBa_J06504. The P1-mCherry plasmid was cloned into a backbone plasmid of Chloramphenicol resistance (pSB1C3).

Usage and Biology

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 7
    Illegal suffix found in sequence at 305
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 7
    Illegal SpeI site found at 306
    Illegal PstI site found at 320
    Illegal NotI site found at 13
    Illegal NotI site found at 313
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 7
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 7
    Illegal suffix found in sequence at 306
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 7
    Illegal XbaI site found at 22
    Illegal SpeI site found at 306
    Illegal PstI site found at 320
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
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