Plasmid_Backbone

Part:BBa_K3712015

Designed by: Yifan Li   Group: iGEM21_HUST2-China   (2021-10-20)


pBV220 plasmid

USAGE AND BIOLOGY

The SD sequence of pBV220 plasmid (sequence used to bind prokaryotic ribosomes) is followed by a polyclonal restriction site, which is convenient for inserting foreign genes with the starting ATG, and can express non-fusion proteins.The vector contains a strong transcription terminator to prevent the "read through" phenomenon, which is conducive to the stability of the plasmid-host system.The vector pBV220 is 3.7Kb and helps increase its copy number and capacity. pBV220 contains the PL promoter and the cI protein gene cIts857, which codes for the inhibitory effect of the promoter and is temperature-sensitive, to regulate the gene pL/pR, so the transcription of the foreign gene inserted into it can be regulated by temperature.

RESULTS

Part:BBa_K3712013

Figure 1 The agarose gel electrophoresis map of Wild-type BLP-7-27 ELP, Optimized BLP-7-27 ELP, Wild-type TLR2-27ELP, and Optimized TLR2-27 ELP respectively constructed in pR/pL expression system. About 200-1000 ng of plasmid was digested at 37℃ for 30-60 minutes and analyzed on 1% Agarose Gel. Lane1: circular plasmid (brighter) and supercoiled structure; Lane2: plasmid (brighter) digested by EcoRI and BamHI and target gene fragment.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 1
    Illegal PstI site found at 40
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 1
    Illegal PstI site found at 40
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 1
    Illegal BglII site found at 3419
    Illegal BamHI site found at 7
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 1
    Illegal PstI site found at 40
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 1
    Illegal PstI site found at 40
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI site found at 220
    Illegal BsaI site found at 1287
    Illegal SapI.rc site found at 2369


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