Reporter system for SST and FFT (BBa_K899000 and BBa_K899001)
Reporter system to test for transcription of fructan fructan fructosyltransferase (FFT) and Sucrose sucrose fructosyltransferase (SST) genes with green flourescent protein (GFP) as reporter, and lacl as promoter
The Plac insert was PCR'd from the MG1655 strain of E.coli K12.
Inverting regulatory region controlled by LacI (<bb_part>BBa_C0010</bb_part>, <bb_part>BBa_C0011</bb_part>, etc.) The pLac regulatory region is a 243 base-pair sequence with standard BioBrick prefix and suffix sections on its ends. It contains two protein binding sites: CAP, which is generally present in E.coli and is assocciated with cell health and availability of glucose., and LacI, the Lac inhibitor <bb_part>BBa_C0010</bb_part> which binds in an dimerized cooperative manner to inhibit the transcription of the protein that follows. In the presence of lactose or IPTG, an analog of lactose, LacI is unable to correctly bind and inhibit transcription. This allows <bb_part>BBa_R0010</bb_part> to be used as a inverter or as a detector of lactose or IPTG.
LacI binds to this regulator. This part is incompatible with species containing active LacI coding regions. Lactose and IPTG disable the operation of LacI and this regulator. This part is incompatible with environments containing lactose or lactose analogs.
Sucrose sucrose fructosyltransferase (SST)
Extracted from Jerusalem artichoke (Helianthus tuberosus).
Catalyzes sucrose to the three products: D-sucrose, beta-D-fructofuranosyl-(2->1) and alpha-D-glucopyranoside, as such: 2 sucrose -> D-glucose + beta-D-fructofuranosyl-(2->1)-beta-D-fructofuranosyl + alpha-D-glucopyranoside.
We removed illigal restriction sites; we added a Shinedalgarno sequence 7 bp from the coding part, to insure expression, while keeping the Kozak sequence to ensure that the part can still be expressed in plants.
Fructan fructan fructosyltransferase (FFT)
Extracted from Jesrusalem Artichoke (helianthus tuberosus)
Elongates a trisaccharide with the fructose unit of a sucrose substrate into an inulin polymer. Works together with part BBa_K899000 (SST).
7 BP from start codon we embedded an shine dalgarno sequence.
Doubble ribosomal binding site (FFT) (shine-dalgarno and kozak sequence)
jerusalem artichoke (Helianthus Tuberosus) and modified with bacterial shine-dalgarno sequence
This is the ribosomal binding site(s) we used in front of our fructan:fructan fructosyltransferase (FFT. It consists of the kozak sequence originally from the gene when we extracted it from the jerusalem artichoke (Helianthus Tuberosus), and the shine dalgarno sequence aggaggt.
the shine dalgarno is placed 6 bp from the startcodon (optimal distance) and kozak is 4 of theese bp right before start codon.
Doubble ribosomal binding site (SST) (shine-dalgarno and kozak sequence)
Kozak sequence derives from jerusalem artichoke (helianthus tuberosus) shinedalgarno was introduced by primers.
This sequence is taken from the RBS section of our sucrose:sucrose fructosyltransferase (SST) gene. It consists of a shine dalgarno sequence (AGGAGGT) and a kozak sequence (CACC).
Derived from the TE terminator of T7 bacteriophage between Genes 1.3 and 1.4 <genbank>V01146</genbank>.
Transcription terminator for the E.coli RNA polymerase.
Suggested by Sri Kosuri and Drew Endy as a high efficiency terminator. The 5' end cutoff was placed immediately after the TAA stop codon and the 3' end cutoff was placed just prior to the RBS of Gene 1.4 (before AAGGAG). Use anywhere transcription should be stopped when the gene of interest is upstream of this terminator.
RBS.3 (medium) -- derivative of BBa_0030
Weak1 RBS based on Ron Weiss thesis. Strength is considered relative to <bb_part>BBa_B0030</bb_part>, <bb_part>BBa_B0031</bb_part>, <bb_part>BBa_B0033</bb_part>.
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix ("RBS-2" in figure 4-14 of thesis). Contact info for this part: Brian Chow
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003.
Transcriptional terminator consisting of a 64 bp stem-loop.
green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (SwissProt: P42212
GFP (mut3b) [note that this part does not have a barcode]
Conversion of the iGEM parts registry to SBOL2.1