Difference between revisions of "Part:BBa K2075022:Design"

Latest revision as of 20:40, 15 October 2016

Vector iGEM_02_Hax3_2xNN

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 2636
1000
COMPATIBLE WITH RFC[1000]

Design Notes

This composite part includes an Intein and linker, an eGFP, a TAL-effector, a TEV Site and a Strep Tag. They translated all together. The intein and linker are important to circularice the aminoacid sequence between the N- and C-terminal parts. Between the Intein sites is an eGFP and the TAL-effector in frame. There is also a strep XT tag and the TEV Site. When it is translated the intein sites react and form a circularised protein. All the parts between N- and C-Terminal Intein is part of the protein circle. In this circle the hole protein gains more stability. With the help of the strep XT tag the protein can be purified even if it is circularised. The Strep XT Tag is also usefull to detect the protein with an immunostain. The TEV Site give the opportunity to linearise the protein again wit the help oft he TEV protease. The linearised TAL can bind the specific DNA Sequence. The eGFP gives us the opportunity to detect the connected TAL-effector. The TAL-effector is Hax3-2xNN. The 12th and 13th amino acid of each of the 11.5 repeats from our TAL Hax3 – 2xNN are: NI HD NI HD HD HD NN NN NS HD NI They bind the DNA sequence: A C A C C C G/A G/A N C A.

Source

Source info goes here

References

References go here

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-This composite part includes an Intein and linker, an eGFP, a TAL-effector, a TEV Site and a Strep Tag. They translated all together. The intein and linker are important to circularice the aminoacid sequence between the N- and C-terminal parts. Between the Intein sites is an eGFP and the TAL-effector in frame. There is also a strep XT tag and the TEV Site. When it is translated the intein sites react and form a circularised protein. All the parts between N- and C-Terminal Intein is part of the protein circle. In this circle the hole protein gains more stability.
-With the help of the strep XT tag the protein can be purified even if it is circularised. The Strep XT Tag is also usefull to detect the protein with an immunostain. The TEV Site give the opportunity to linearise the protein again wit the help oft he TEV protease. The linearised TAL can bind the specific DNA Sequence. The eGFP gives us the opportunity to detect the connected TAL-effector. The TAL-effector is Hax3-2xNN. The 12th and 13th amino acid of each of the 11.5 repeats from our TAL Hax3 – 2xNN are: NI HD NI HD HD HD NN NN NS HD NI They bind the DNA sequence: A C A C C C G/A G/A N C A.
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