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Team:UT-Tokyo/Sudoku lab note
From 2010.igem.org
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Revision as of 03:04, 19 September 2010
Sudoku
Abstract Construct Lab note Result
Lab note
This is an abstract page. If you can see detail data, please click each protocols.
Main construct
Detail protocols about this is:
June.
July.
Assay
Terminator leak switch
To realize 4C3 leak switch, we choose proper terminator which terminates transcription when connected two or more but leak when single.
We made assay (see Fig) to select proper terminator.
flpe
The original flpe include two restriction enzyme sites(EcoRI, SpeI).
We modified this flpe by PCR:
1st -> cloning
2nd -> modified SpeI site
3rd -> modified EcoRI site
Convert lox71(rev) to lox66(rev)
Convert lox71(rev) to lox66(rev)
Make Hin(rev)
Virus expression check
Anti-RNA repression
Parts list
| Number | Name | Link to BioBrick | Plate coordinate | Vector | Code length |
| 1 | T7 promoter |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_I712074 BBa_I712074] | plate1-6N | pSB1AK8 | 46bp |
| 2 | rbs |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_B0030 BBa_B0030] | plate1-1H | pSB1A2 | 15bp |
| 3 | cre recombinase |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_J61047 BBa_J61047] | plate1-5D | pSB1A2 | 1037bp |
| 4 | double terminator |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_B0014 BBa_B0014] | plate2-24C | pSB1AK3 | 95bp |
| 5 | lox66 recombinase site |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_I718017 BBa_I718017] | plate1-17J | pSB1A2 | 34bp |
| 6 | Kan resistance (rev) |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_J31002 BBa_J31002] | plate1-2K | pSB1A2 | 816bp |
| 7 | rbs (rev) |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_B0014 BBa_B0014] | plate1-1J | pSB1A2 | 15bp |
| 8 | lox71 recombinase site |
order-made | --- | --- | 34bp |
| 9 | single terminator |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_B1006 BBa_B1006] | plate1-4H | pSB1AK3 | 39bp |
| 10 | constant express promoter |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_J23119 BBa_J23119] | plate1-18A | pSB1A2 | 35bp |
| 11 | Tet resistance (rev) |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_J31006 BBa_J31006] | plate1-1N | pSB1A2 | 1191bp |
| 12 | hixC |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_J44000 BBa_J44000] | plate1-1B | pSB1A2 | 26bp |
| 13 | --- |
--- | --- | --- | --- |
| 14 | lox66 |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_I718016 BBa_I718016] | plate1-17H | pSB1A2 | 34bp |
| 15 | rbs-mRFP1-terminator |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_I13507 BBa_I13507] | plate1-22O | pSB1A2 | 861bp |
| 16 | location sequence(old) |
order | --- | --- | --- |
| 17 | pSP6(rev) |
order | --- | --- | --- |
| 18 | lox2272 |
order | --- | --- | --- |
| 19 | loading sequence(rev) |
order | --- | --- | --- |
| 20 | frt |
[http://partsregistry.org/Part:BBa_J61020 BBa_J61020] | From HQ | pSB1A2 | 34bp |
| 21 | hin(true) |
[http://partsregistry.org/Part:BBa_J31000 BBa_J31000] | From HQ | ??? | 573bp |
| 22 | hin(rev) |
--- | --- | ??? | 573bp |
| 23 | flpe |
--- | --- | --- | about 1.2kbp |
| 24 | gfp unit |
--- | --- | --- | about 1.2kbp |
| 25 | flpe(rev) |
--- | --- | --- | about 1.2kbp |
