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Team:Paris Bettencourt/Notebook/Eliminate Smell
From 2014.igem.org
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| - | <p> | + | <p>To amplify this construct, we created two oligos:</p> |
| - | + | <TABLE BORDER> | |
| + | <TR><b><TD>Name</TD><TD>Sequence (5'->3')</TD><TD>Notes</TD><TDBinding position</b></TD></TR> | ||
| + | <TR><TD><b>oPB.001</b></TD><TD>TATAGAATTCGCGGCCGCTTCTAGAGTGACAGCTAGCTCAGTCCTAGG</TD><TD>agaA forward for BioBrick vector</TD><TD>1->23</TD></TR> | ||
| + | <TR><TD><b>oPB.002-BAD PRIMER</b></TD><TD>GAAGCATCATCACCATCACCACTGATACTAGTAGCGGCCGCTGCAGTTA</TD><TD>agaA REVERSE for BioBrick vector- NOT REVERSED</TD><TD>1272->1296</TD></TR> | ||
| + | <TR><TD><b>oPB.005</b></TD><TD>TTAACTGCAGCGGCCGCTACTAGTATCAGTGGTGATGGTGATGATGCTTC</TD><TD>agaA reverse for BioBrick vector</TD><TD>1272->1296</TD></TR> | ||
| + | </TABLE> | ||
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<h5>Date 2</h5> | <h5>Date 2</h5> | ||
<p>Text</p> | <p>Text</p> | ||
Revision as of 14:13, 19 August 2014
Paris Bettencourt 2014
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Eliminate Smell
Notebook
June
June 23rd
Goal: Design plasmids that express agaA construct
Procedure:
Using Geneious, we first created the agaA construct:
- Promoter BBa_J23108 from the Anderson's promoter collection
- RBS from the RBS Calculator of Salis lab specific for E coli
- BioBrick Prefix + Scar from the iGEM Parts webpage
- agaA sequence codon optimized for E coli 12 (IDT online tool). We checked with Generious that there were no restriction sites for EcoRI, SpeI, ZbaI and PstI.
- Histidine tag: 6 Histidines in C-terminal were added
- Stop codon
- BioBrick scar + BioBrick suffix from the iGEM Parts webpage
To amplify this construct, we created two oligos:
| Name | Sequence (5'->3') | Notes | |
| oPB.001 | TATAGAATTCGCGGCCGCTTCTAGAGTGACAGCTAGCTCAGTCCTAGG | agaA forward for BioBrick vector | 1->23 |
| oPB.002-BAD PRIMER | GAAGCATCATCACCATCACCACTGATACTAGTAGCGGCCGCTGCAGTTA | agaA REVERSE for BioBrick vector- NOT REVERSED | 1272->1296 |
| oPB.005 | TTAACTGCAGCGGCCGCTACTAGTATCAGTGGTGATGGTGATGATGCTTC | agaA reverse for BioBrick vector | 1272->1296 |
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July
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August
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September
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October
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