Part:BBa_K1789013
SCAF1+Ter
This is a combination of scaffold and termination.
Usage and Biology
Usage: This part is scaf1 which is associated with a terminator. Scaf1 is used to bind TALE1 and TALE3 protein to make them fixed on the DNA. We create this part to make it more convenient every time we are required to add a scaffold.
Biology:
Scaf1 is a DNA scaffold which can bind TALE protein specifically to achieve the fixing of protein or enzyme. This part is scaf1 with a terminator after it.
Sequence and Features
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 145
Illegal AgeI site found at 175
Illegal AgeI site found at 205
Illegal AgeI site found at 235
Illegal AgeI site found at 265
Illegal AgeI site found at 295
Illegal AgeI site found at 325 - 1000COMPATIBLE WITH RFC[1000]
Experimental Validation
This part is validated through four ways: Enzyme cutting, PCR, and Sequence
PCR
Methods
The PCR is performed with Premix EX Taq by Takara.
The PCR protocol is selected based on the Users Manuel. The Electrophoresis was performed on a 1％ Agarose glu.
Results
The result of the agarose electrophoresis was shown on the picture above.
Enzyme cutting
Methods
After the assembly ,the plasmid was transferred into the Competent E. coli DH5α). 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 EcoRI and XbaI restriction endonuclease bought from TAKARA.
The plasmid was cutted in a 20μL system 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 picture above.
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