Difference between revisions of "Part:BBa K2235009"
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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. | 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. | ||
− | [[File:SiaComp_1.png|600px|thumb|left|Figure 1]] | + | [[File:SiaComp_1.png|600px|thumb|left|Figure 1: |
+ | From left to right: M DNA ladder, next 2 lanes are plasmids digested with Ecor1 and Pst1.]] | ||
− | |||
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Revision as of 15:25, 26 October 2017
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.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 127
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 574
Illegal NgoMIV site found at 649
Illegal NgoMIV site found at 739 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1119