Difference between revisions of "Part:BBa K797004"

Latest revision as of 14:00, 25 September 2012

Tat secretion cassette with constitutive promoter.

This cassette allow E.coli to secrete proteins with torA signal. Wild type Tat protein secretion system is too week so that Kyoto 2012 construct Tat cassette to reinforce the ability of transportation of Tat system. This parts include tatA,B,C protein coding region and pspA (phage shock protein A). Tat A,B,C protein is the main component of Tat complex where proteins with torA signal go through and pspA can encourage protein secretion via Tat system. Kyoto 2012 suggest iGEMers with this new way of secretion and provide them with this cassette regulated by constitutive promoter.
We checked the sequence of tatABCD(BBa_K797000) and the sequence of pspA(BBa_K797001) individually, and then, we made TAT construction composed of constitutive promoter(BBa_J23107), tatABCD(BBa_K797000),pspA(BBa_K797001) and double terminator(BBa_B0015). This TAT secretion cassette is too long device to sequence, so that we performed electrophoresis of this cassette and confirmed the length of our parts.

Considering that the sequences of tatABCD and pspA are correct,and the length of TAT secretion casssette is correct, we declare that this construction of TAT secretion cassette has been completed.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 2621
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 2291
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 2048
Illegal SapI.rc site found at 2081

@@ Line 5: / Line 5: @@
 This parts include tatA,B,C protein coding region and pspA (phage shock protein A). Tat A,B,C protein is the main
 component of Tat complex where proteins with torA signal go through and pspA can encourage protein secretion via Tat system. Kyoto 2012 suggest iGEMers with this new way of secretion and provide them with this cassette regulated by constitutive promoter.<br>
-We checked the sequence of tatABCD[[Part:BBa_K797000|'''(BBa_K797000)''']]and the sequence of pspA(BBa_K797001　このページへリンク) individually, and then, we made TAT construction composed of constitutive promoter(BBa_J23107　このページへリンク), tatABCD(BBa_K797000　このページへリンク), pspA(BBa_K797001　このページへリンク) and double terminator(BBa_B0015　このページへリンク). This TAT secretion cassette is too long device to sequence, so that we performed electrophoresis of this cassette and confirmed the length of our parts. <br><br>
+We checked the sequence of tatABCD[[Part:BBa_K797000|'''(BBa_K797000)''']] and the sequence of pspA[[Part:BBa_K797001|'''(BBa_K797001)''']] individually, and then, we made TAT construction composed of constitutive promoter[[Part:BBa_J23107|'''(BBa_J23107)''']], tatABCD[[Part:BBa_K797000|'''(BBa_K797000)''']],pspA[[Part:BBa_K797001|'''(BBa_K797001)''']] and double terminator[[Part:BBa_B0015|'''(BBa_B0015)''']]. This TAT secretion cassette is too long device to sequence, so that we performed electrophoresis of this cassette and confirmed the length of our parts. <br><br>
-Considering that the sequences of tatABCD and pspA are correct, and the length of TAT secretion casssette is correct, we declare that this construction of TAT secretion cassette has been completed.
+Considering that the sequences of tatABCD and pspA are correct,and the length of TAT secretion casssette is correct, we declare that this construction of TAT secretion cassette has been completed.