Difference between revisions of "Part:BBa K2235009"
Line 3: Line 3:
 
<partinfo>BBa_K2235009 short</partinfo>
 
<partinfo>BBa_K2235009 short</partinfo>
  
 +
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.
 +
 +
[[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.
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 18:47, 25 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.

Figure 1

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

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