RBS + Circular λ-Lysozyme with His6 linker
Constantin Ahlmann-Eltze, Charlotte Bunne, Magdalena Büscher, Jan Gleixner, Max Horn, Anna Huhn, Nils Klughammer, Jakob Kreft, Elisabeth Schäfer, Carolin Schmelas, Silvan Schmitz, Max Waldha
This is the sequence of the SpeI/XbaI scar for RBS-CDS junctions in BioBricks standard assembly.
This is a shorter scar to ensure proper spacing between the RBS and CDS.
synthesized as found in the T7 genome and several commercial expression plasmids.
RFC10 compatible strong RBS derived from the T7 phage gene 10a (major capsid protein)[1]. When assembled to a coding part with the A of the start codon being part of the XbaI site, the RBS will be shifted one bp downstream compared to the native sequence. The sequence was successfully used by the iGEM team Heidelberg 2014 for the expression of many proteins in E.coli. 1. Olinss, P. & Rangwala, S. H. Derived from Bacteriophage T7 mRNA Acts ELS an Enhancer of Translation of the lac2 Gene in. 16973???16976 (1989).
The 18 bp including the XbaI that can be found upstream of the presumably important part of the RBS were included into the sequence just to make sure it works. However to fully comply with RFC10 a G was inserted behind the XbaI site.
Constantin Ahlmann-Eltze, Charlotte Bunne, Magdalena B??scher, Jan Gleixner, Max Horn, Anna Huhn, Nils Klughammer, Jakob Kreft, Elisabeth Sch??fer, Carolin Schmelas, Silvan Schmitz, Max Waldha
N-terminal start overhang (T)(A)-(G)ATG=RBS+Start RFC[105] A
This standard overhang part was developed as part of the iGEM team Heidelberg 2014's Intein Toolbox [[#References[1]]]. All standard sequences can be reviewed in RFC[???] [[#References[2]]].
This is a standard overhang sequence for in-frame cloning of Proteins of Interest in front or behind an Intein. A detailed cloning strategy is found on the iGEM team Heidelberg 2014's wiki page as well as in RFC[???]. Specifically, this part contains the overhang A used to insert a protein behind any RFC[10] compatible RBS and in-frame with the start-codon. It lies within the four bases formed by the second last base of the XbaI/SpeI scar or the first base in front of start codon respectively and the start codon itself.
This part is only the sequence of a standard overhang. It will not be sent in as physical DNA and was merely created to easily compose new parts in the RFC[???] standard. In the actual cloning process this sequence was and is recommended to be inserted with the according PCR primers.
Obtained from pVS41 by Prof. Henning D. Mootz, University of Muenster.
Nostoc punctiforme DnaE split Intein C-terminal half. This is a DNA piece for cloning used to assemble our other BioBrick parts.
This part represents only the intein sequence without inluding the standard splicing-site or polyglycine-linker overhangs.
C-terminal splicing overhang CAAC=1/3?+Asn RFC[105] standard overhang CC
This standard overhang part was developed as part of the iGEM team Heidelberg 2014's Intein Toolbox [[#References|[1]]]. All standard sequences can be reviewed in RFC[???] [[#References|[2]]].
This is a standard overhang sequence for in-frame cloning of Proteins of Interest in front or behind an Intein. A detailed cloning strategy is found on the iGEM team Heidelberg 2014's [http://2014.igem.org/Team:Heidelberg/parts wiki page] as well as in RFC[???]. Specifically, this part contains the overhang E used to insert a protein in front of an N-Intein. It lies within the four bases formed by the cystein and the first base of the subsequent Leucine/Isoleucine or similar of the N-terminal splicing site.
Obtained from pVS07 by Prof. Henning D. Mootz, University of Muenster.
Nostoc punctiforme DnaE split Intein N-terminal half. This is a DNA piece for cloning used to assemble other BioBrick parts.
N-terminal splicing overhang TGCT=Cys+1/3 Leu|Ile RFC[105] overhang NN
C-terminal stop overhang TAAT=STOP+1/3Stop RFC[105] overhang Z
This is a standard overhang sequence for in-frame cloning of Proteins of Interest in front or behind an Intein. A detailed cloning strategy is found on the iGEM team Heidelberg 2014's [http://2014.igem.org/Team:Heidelberg/parts wiki page] as well as in RFC[???]. Specifically, this part contains the overhang F used to insert the last part of an intein-fused protein directly in front of an RFC[10] double stop-codon. It lies within the first for bases of the stopstop sequence.
Directly synthesized as an oligo as backtranslated with EMBOSS[1]. ==References== 1. Rice, P., Longden, I. & Bleasby, A. EMBOSS: the European Molecular Biology Open Software Suite. Trends Genet. 16, 276???7 (2000).
This Part adds ultimate coolness to you your part. More seriously, it served us as a random sequence being a better overlap site for CPEC cloning than the RFC[10] suffix.
We aimed for a sequence without secondary structures, balanced GC content.
Conversion of the iGEM parts registry to SBOL2.1