Template:SBB12 part sbb1227: Difference between revisions
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<pre> | <pre> | ||
Partname: sbb1227 | Partname: sbb1227 | ||
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Just a big hydrophobic patch of 8 leucines, or LLLLLLLL* | Just a big hydrophobic patch of 8 leucines, or LLLLLLLL* | ||
{{SBB12_homodimerizer}} | {{SBB12_homodimerizer}} | ||
<blockquote style="background: white; border: 1px solid rgb(117, 174, 255); font: typewriter; padding: 2em;" | |||
<blockquote style="background: white; border: 1px solid rgb(117, 174, 255); font: typewriter; padding: 2em;"> | |||
===Construction File=== | ===Construction File=== | ||
<pre> | |||
</ | Wobble tn001F/tn002R (64bp, wobpdt) | ||
Digest wobpdt (NheI/BamHI, L, wobdig) | |||
Digest pBca9525-Bca1834 (NheI/BamHI, L, vectdig) | |||
Ligate wobdig and vectdig (pBca9525-sbb1227) {Leu8} | |||
---------------------------------------------------------------------- | |||
>tn001F Forward construction of sbb1227 basic part | |||
ccataGCTAGCggcagtggatctggtCTGCTACTGTTGCTG | |||
>tn002R Reverse construction of sbb1227 basic part | |||
CTGATggatccTTACAGGAGTAGCAGCAACAGTAGCAGaccag | |||
>wobpdt | |||
ccataGCTAGCggcagtggatctggtCTGCTACTGTTGCTGCTACTCCTGTAAggatccATCAG | |||
</pre></blockquote> | |||
__NOTOC__ | __NOTOC__ |
Latest revision as of 19:38, 13 February 2012
Partname: sbb1227 Featurename: Leu8 Genename: n/a Source: Synthetic
Just a big hydrophobic patch of 8 leucines, or LLLLLLLL*
This part encodes a homodimer domain
You are going to put the homodimer on the C-terminus of a truncated ToxR. For this part, you're going to go off BioBricks. Kindov. Your final product will be a BioBrick, but you are going to use an ad hoc procedure to create it. You should examine this illustration which refers to two sequences: pBca9525-Bca1834 and pBca9525-Bca1839. The important thing here is that you think through the translation frame of the final product, and make sure that you add some linker between your sequence and the sequence 5' to it. You should also remove the start methionine from you homodimer, unless your project description explicitly says not to. You should include the stop codon.
The product of your construction file should resemble pBca9525-Bca1839 in terms of it encoding a full expression cassette containing a ToxR fusion protein with your designed feature. If you were given part sbb1293, your product would be called pBca9525-sbb1293.
Construction File
Wobble tn001F/tn002R (64bp, wobpdt) Digest wobpdt (NheI/BamHI, L, wobdig) Digest pBca9525-Bca1834 (NheI/BamHI, L, vectdig) Ligate wobdig and vectdig (pBca9525-sbb1227) {Leu8} ---------------------------------------------------------------------- >tn001F Forward construction of sbb1227 basic part ccataGCTAGCggcagtggatctggtCTGCTACTGTTGCTG >tn002R Reverse construction of sbb1227 basic part CTGATggatccTTACAGGAGTAGCAGCAACAGTAGCAGaccag >wobpdt ccataGCTAGCggcagtggatctggtCTGCTACTGTTGCTGCTACTCCTGTAAggatccATCAG