Composite

Part:BBa_K2235009

Designed by: Shivashree Dhanaraj, Shanlin Tong, Gilai Nachmann and Sina Amoor pour   Group: iGEM17_Stockholm   (2017-10-11)
Revision as of 15:37, 26 October 2017 by Tongshl (Talk | contribs)


Sialidase composite with T7 promoter and RBS

Sialidase enzyme coding sequence with T7 promoter and RBS

Molecular cloning

Ligation of sialidase composite insert into pSB1C3

The gblock containing T7 promoter-RBS-sialidase was ligated into the Chloramphenicol plasmid backbone (pSB1C3). Preliminary confirmation of cloning was done by double digest Ecor1 and Pst1. Figure 1 represents the double digestion result: bands were visible at ~1600 bp and ~2000 bp which represents the insert and plasmid backbone respectively.

Figure 1: From left to right: M DNA ladder, next 2 lanes are plasmids digested with Ecor1 and Pst1.

Important Parameters

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 127
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 574
    Illegal NgoMIV site found at 649
    Illegal NgoMIV site found at 739
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1119


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Categories
Parameters
None