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Part:BBa_K1532001

Designed by: Xinyuan Qiu   Group: iGEM14_NUDT_CHINA   (2014-09-30)

Sal I cutting site

This part is the cutting site of the restriction endonuclease Sal I. The endonuclease Sal 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

The enzyme Sal I is a restriction endonuclease found in Streptomyces albus. It can specifically recognize the sequence

...GTCGAC...

...CAGCTG...

And cut it into

...G TCGAC...

...CAGCT G...

At the condition of 37 degrees Celsius.

SalI_DES.jpg

In this part, we standardized the cutting sequence of the Sal I so that a new cutting site can be added into a system conveniently to leave the system opened for further edit.

SalI_DIS2_300_450.jpg

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 Sal 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_K1532001 can be cut by the Sal I enzyme under the given circumstances.

SalItest.jpg

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
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Parameters
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