Part:BBa_K2814003
pLTL+RBS+sfGFP+ssrA deg tag+ Terminator
This part was formed by ligating hybrid promoter(lac-tet-lac) with the a reporter sequence containing superfolder GFP along with a ssrA degradation tag and a terminator, a RBS preceded this sequence so that the hybrid promoter could be used in tandem. Hence, sfGFP can be induced by IPTG or/and aTc depending upon the repressors acting upon it.
The pLTL (Lac-Tet-Lac) (BBa_K2814001) is a hybrid promoter. It is composed of the operator and promoter sequences of pTet and pLac promoter. There are multiple benefits of using the pLTL promoter:-
1) The hybrid pLTL shows almost complete repression on being repressed and on complete induction (either by IPTG or aTc), the hybrid pLTL promoter shows a 1.4-2 times higher expression. 2) The hybrid pLTL promoter also permits flexible gene expression because it can be utilized under either or both repression controls (LacI and TetR) simultaneously.
The RBS used is standard Elowitz RBS and we have use a codon optimized(for E. coli) variant of superfolder GFP along with a ssrA degradation tag in order to speed up the degradation rate of the GFP. More information related to the sfGFP+ssrA degtag and the part sequence can be found on it’s BioBrick page (BBa_K2814007).The terminator used is a standard double terminator composed of rrnb T1 and T7 Te terminators. (BBa_K2814008)
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 142
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