Histidine tag and RFP linker [GSGSGS*2] semi-permanent generator
This device is all-in-one mRNA circularization device. If you transfer this device into E. coli, you can get a protein (its sequence is ???GSGSGSGSGSGS - amino acid derived from RBS sequence - HisRFP ??? GSGSGSGSGSGS - amino acid derived from ribozyme sequence ??? GSGSGSGSGSGS - amino acid derived from RBS sequence -???)
This part includes sequences which one of the linkers we designed(GSGSGSGSGSGS).
RBS (Elowitz 1999) -- defines RBS efficiency
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix. No secondary structures are formed in the given RBS region. Users should check for secondary structures induced in the RBS by upstream and downstream elements in the +50 to -50 region, as such structures will greatly affect the strength of the RBS. Contact info for this part: Brian Chow
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003.
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.
Double terminator consisting of BBa_B0010 and BBa_B0012
the 3??side of the intron[BBa_K1332005] +GSGSGS*2 linker
This part consists of mRNA circularization device (5?? side) and linker (amino acid sequence: GSGSGSGSGSGS). It???s the improved mRNA circularization parts (5?? side). The linker may keep a function of coded protein that is translated semi-permanently. The amino acids that consist of the linker is glycine and serine. Because their steric hindrance is small and their hydrophilicity are strong. The protein coding sequence that is inserted between this device and mRNA circularization device (3??? side) can be circularized. If you circularized the protein coding sequence (Its a stop codon have been removed.) with mRNA circularization device (3?? side) (endless translation)(BBa_K1332009), you can get a circular mRNA that is translated semi-permanently.
We built a sequence of linker(GSGSGSGSGSGS) into the circuler mRNA. The linker may keep a function of coded protein that is translated semi-permanently.
2*GSGSGS linker+ the 5??side of the intron[BBa_K1332003]
This part consists of mRNA circularization device (3?? side) (endless) and linker (amino acid sequence: GSGSGSGSGSGS). It???s the improved mRNA circularization parts (5?? side). The linker may keep a function of coded protein that is translated semi-permanently. The amino acids that consist of the linker are histidine. Because it has some steric hindrance but its hydrophilicity is very strong. The protein coding sequence that is inserted between this part and mRNA circularization device (5??? side)(BBa_K1332008) can be circularized. If you circularized the protein coding sequence (Its stop codon have been removed.), you can get a circular mRNA that is translated semi-permanently.
We built a sequence of linker (GSGSGSGSGSGS) into the circuler mRNA. The linker may keep a function of coded protein that is translated semi-permanently.
Histidine tag (8 AA) and RFP (without stop codon)
This part consists of histidine tag and RFP. His-tag is useful for purifying a long RFP. The RFP lacks its stop codon.
Stop codons are removed to translate consecutively.
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.
Transcriptional terminator consisting of a 64 bp stem-loop.
Conversion of the iGEM parts registry to SBOL2.1