Plasmid

Part:BBa_K5082009

Designed by: Fangyuan Duan   Group: iGEM24_YiYe-China   (2024-09-07)

Usage and Biology

Plasmids are double stranded circular DNA molecules, which carry non-essential genes, found in bacterial cells [1]. Plasmids are multifunction, easy to edit, and can be amplified quickly by bacterial cellular machinery [2]. These advantages have made plasmids a popular tool in genetic research.

pHAGE is a mammalian expression vector [3]. The pHAGE plasmid can be amplified by DH5α bacterial cells. Meanwhile, it carries an ampicillin resistance gene for selection. The plasmid map for pCMV-EGFP is shown in Figure 1. 


         phage.png
            Figure 1. pHAGE plasmid map.

Design

The above characteristics have made pHAGE an ideal plasmid for our experiment. In our experiment, we inserted a G3BP1 gene into the plasmid. Upon amplification, we transfected the plasmid into stomach cells at different concentrations to measure the sensitivity of the sensor we designed. The results were as expected, proving the effectiveness of this plasmid backbone.


Reference

[1] Carattoli, Alessandra. “Plasmids in Gram Negatives: Molecular Typing of Resistance Plasmids.” International Journal of Medical Microbiology, vol. 301, no. 8, Dec. 2011, pp. 654–658

[2] Patron, N.J. Synthetic Biology and Gene Cloning. Elsevier EBooks, Elsevier BV, 1 Jan. 2017, pp. 112–117

[3] “Addgene: PTS106 PHAGE2 CMVtetO2 MCS-3C-SUMOEu-22xGS-GFP.” Addgene.org, 2024, www.addgene.org/199359/. Accessed 15 Aug. 2024.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2028
    Illegal EcoRI site found at 2972
    Illegal SpeI site found at 2212
    Illegal SpeI site found at 3227
    Illegal PstI site found at 3484
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2028
    Illegal EcoRI site found at 2972
    Illegal SpeI site found at 2212
    Illegal SpeI site found at 3227
    Illegal PstI site found at 3484
    Illegal NotI site found at 2879
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2028
    Illegal EcoRI site found at 2972
    Illegal BglII site found at 473
    Illegal BglII site found at 4743
    Illegal BglII site found at 4809
    Illegal BglII site found at 4850
    Illegal BamHI site found at 2859
    Illegal XhoI site found at 2871
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2028
    Illegal EcoRI site found at 2972
    Illegal SpeI site found at 2212
    Illegal SpeI site found at 3227
    Illegal PstI site found at 3484
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2028
    Illegal EcoRI site found at 2972
    Illegal SpeI site found at 2212
    Illegal SpeI site found at 3227
    Illegal PstI site found at 3484
    Illegal NgoMIV site found at 1146
    Illegal NgoMIV site found at 4627
    Illegal AgeI site found at 3092
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 455
    Illegal BsaI site found at 4832
    Illegal BsaI site found at 6036
    Illegal BsaI.rc site found at 3982
    Illegal SapI.rc site found at 1107
    Illegal SapI.rc site found at 7118


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