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Revision as of 14:52, 23 October 2007 by Stefan Luzi (Talk | contribs)

System Parts

educatETH E.coli consists of 11 parts that can be synthesized independently (want to know how this is done in the lab? Then visit our In the Lab page!) Please note that four of them (4,5 and 8,9) form together two functional system units. They have been separated to ensure comparable part lengths and thus enable easier introduction into plasmids. All parts are listed in the following:

Single System Parts
1 TetR production BBa_I739001 constitutive subsystem
2 LacI production BBa_I739002 constitutive subsystem
3 LuxR production BBa_I739003 constitutive subsystem
4 1st half of P22 cII / EYFP production BBa_I739004 reporting subsystem
5 2nd half of P22 cII / EYFP production BBa_I739005 reporting subsystem
6 cI production BBa_I739006 learning subsystem
7 P22 cII production BBa_I739007 learning subsystem
8 1st half of cI / ECFP production BBa_I739008 reporting subsystem
9 2nd half of cI / ECFP production BBa_I739009 reporting subsystem
10 RFP production BBa_I739010 reporting subsystem
11 GFP production BBa_I739011 reporting subsystem

Composite System Parts
1+2+3 tetR + lacI + luxR production BBa_I739013 constitutive subsystem
4+5 P22 cII + EYFP production BBa_I739015 reporting subsystem
8+9 cI + ECFP production BBa_I739016 reporting subsystem
(4+5)+(8+9) (P22 cII + EYFP) + (cI + ECFP) production BBa_I739017 reporting subsystem
6+7 cI + P22 cII production BBa_I739018 learning subsystem
10+11 RFP + GFP production BBa_I739019 reporting subsystem
(6+7)+(10+11) (cI + P22 cII) + (RFP + GFP) production BBa_I739020 learning/reporting subsystem

Many of the above mentioned parts contain double promoters. This promoter constructs contain two independent classes of operator sites and can therefore be regulated by two different types of molecules. Double promoters form the basis of the inducible toggle switch and hence the memory of our learning system. This concept could also help future projects in developing devices and systems that need extended regulation. In the following, we introduce a selection of first generation double promoters to the Registry of Standard Biological Parts:

Double Promoters
1' cI negative / tetR negative promoter BBa_I739102 reporting subsystem
2' lacI negative / P22 cII negative promoter BBa_I739103 reporting subsystem
3' luxR/HSL positive / P22 cII negative promoter BBa_I739104 learning subsystem
4' luxR/HSL positive / cI negative promoter BBa_I739105 learning subsystem
5' tetR negative / P22 cII negative promoter BBa_I739106 reporting subsystem
6' cI negative / lacI negative promoter BBa_I739107 reporting subsystem

In order to test if the concept of those proposed double promoters is working, simple proof of concept (PoC) parts have been constructed. The PoC promoter consists of two TetR operator sequences linked to a constitutive promoter. In contrast to the other double promoters, this one is only single regulated. The PoC intermediate is part of the PoC composite part, whith which one can test the concept (cI and EYFP production with and without addition of TetR).

Proof of Concept
1" PoC promoter BBa_I739101 proof of concept, no part of the system
2" PoC intermediate BBa_I739014 proof of concept, no part of the system
3" PoC composite BBa_I739021 proof of concept, no part of the system

1"' pBR322BB1 BBa_I739201 high-copy cloning vector, ApR
2"' pCK01BB1 BBa_I739202 low-copy cloning vector, CmR
3"' pACYC177BB1 BBa_I739203 low-copy cloning vector, KmR