Part:BBa_K1532002
Kpn I cutting site
This part is the cutting site of the restriction endonuclease Kpn I. The endonuclease Kpn I can specifically recognize the sequence of this part and cut the double helix into two cohesive ends. You can use this part when you are trying to add an extra cutting site inside of your system in order to make it open-edited.
Part Description
Usage and Biology
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The enzyme Kpn I is a restriction endonuclease found in Klebsiella pneumoniae . It can specifically recognize the sequence
In this part, we standardized the cutting sequence of the Kpn I so that a new cutting site can be added into a system conveniently to leave the system opened for further edit. |
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Cutting and Agarose Electrophoresis
Methods
After the assembly ,the plasmid was transferred into the Competent E. coli DH5α(which is dam+). After culturing overnight in LB,we minipreped the plasmid for cutting. The preparation of the plasmid was performed with TIANprep Mini Plasmid Kit from TIANGEN. The cutting procedure was performed with Kpn I restriction endonuclease bought from TAKARA.
The plasmid was cutted in a 20μL mixture at 37 ℃ for 2 hours. The Electrophoresis was performed on a 1％ Agarose glu.
Results
The result of the agarose electrophoresis was shown on the following picture. As is shown, the plasmid contains BBa_K1532002 can be cut by the Kpn I enzyme under the given circumstances.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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