Plasmid

Part:BBa_K5306007

Designed by: Iulia Beres   Group: iGEM24_MSP-Maastricht   (2024-09-30)

pSEVA261

This basic part encodes for the pSEVA261 plasmid that was used for the expression of the ANRA pathway in Vibrio natriegens.

Usage and Biology

pSEVA261 has a Kanamycin resistance gene and a p15A origin of replication. This plasmid has been shown to be compatible with our chassis organism V. natriegens . According to Tschirhart et al. (2019), pSEVA261 has a low-copy number and high-maintanance levels after 24h. This information is essential as our project aims to express three enzymes constitutively while minimizing the metabolic burden on the organism. This plasmid can serve as a backbone for projects that are interested in expressing multiple genes within a single-plasmid framework. pSEVA261 was obtained from the SEVA Collection.



Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 56
    Illegal XbaI site found at 83
    Illegal SpeI site found at 145
    Illegal PstI site found at 95
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 56
    Illegal SpeI site found at 145
    Illegal PstI site found at 95
    Illegal NotI site found at 46
    Illegal NotI site found at 113
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 56
    Illegal BamHI site found at 77
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 56
    Illegal XbaI site found at 83
    Illegal SpeI site found at 145
    Illegal PstI site found at 95
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 56
    Illegal XbaI site found at 83
    Illegal SpeI site found at 145
    Illegal PstI site found at 95
    Illegal NgoMIV site found at 1475
    Illegal AgeI site found at 1913
  • 1000
    COMPATIBLE WITH RFC[1000]

References

  1. Tschirhart, T., Shukla, V., Kelly, E. E., Schultzhaus, Z., NewRingeisen, E., Erickson, J. S., Wang, Z., Garcia, W., Curl, E., Egbert, R. G., Yeung, E., & Vora, G. J. (2019). Synthetic Biology Tools for the Fast-Growing Marine Bacterium Vibrio natriegens. ACS Synthetic Biology, 8(9), 2069–2079. https://doi.org/10.1021/acssynbio.9b00176
  2. D3veloperSCS_SEVA. (n.d.). Find your plasmid. SEVA Plasmids - Standard European Vector Architecture. https://seva-plasmids.com/canonical-seva-plasmid-list/
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Categories
Parameters
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