tetP promoter: : BBa_R0040 RBS: BBa_B0030 GFP: BBa_E0040 stop: BBa_B0015
mioC promoter + RBS + tetR (standard 10 compatible)
mioC: Escherichia coli p0305293.5 gecp03052935.contig.229, whole genome shotgun sequence. RBS + tetR: BBa_I15040
The MioC promoter is a cell-cycle dependent promoter. Nathan knows this!!! he will modified
tet R coding sequence combined with mioC promoter to allow production of tet R protein during cell replication.
loXf: BBa_J61046 tetP promoter reverse: BBa_R0040 RBS: BBa_B0030 cre: BBa_J61047 ssRa: BBa_K313002 stop: BBa_B0015 loXr: BBa_J61046
Lambda lysis cassette[BBa_K358004] with double terminator[BBa_B0015].
RBS.1 (strong) -- modified from R. Weiss
Strong RBS based on Ron Weiss thesis. Strength is considered relative to <bb_part>BBa_B0031</bb_part>, <bb_part>BBa_B0032</bb_part>, <bb_part>BBa_B0033</bb_part>.
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix ("orig" in figure 4-14 of Ron Weiss thesis). 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 Brian Chow
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003.
Double terminator consisting of BBa_B0010 and BBa_B0012
This is the naturally-occurring (possibly stationary-phase only) promoter for the XylR gene (which encodes the transcriptional regulator XylR).
This is the promoter activated by the transcriptional regulator XylR (BBa_I723136) in response to detection of environmental xylene.
AANDENYNYADAS. (Fast) SsrA degradation tag.
This variation of the SsrA tag was studied by McGinness, Baker, Sauer. 2006. Mol. Cell. 22:701.
This sequence codes for the amino acid sequence AANDENYNYADAS, a variation of the WT SsrA tag sequence, which, when fused to the C-terminal of proteins, will make the protein susceptible to fast degradation through SspB-mediated binding to the ClpX protease. The following rates of degradation of this tag are pulled from the corresponding references below: ~2 Vmax/ [Clpx6] min-1 from (1). See the following references for further information on degradation rates and mechanisms of this tag: (1)McGinness, Baker, Sauer. 2006. Mol. Cell. 22:701. (2)Flynn et al 2003. Mol. Cell. 11: 671. Flynn et al. 2001. PNAS 98(19): 10584. Anderson et al 1998. App. Env. Microbiol. 64(6):2240.
C-terminal tag. Degradation rate is fast. Variations of this sequence yield different degradation rates(see Parts BBa_M0050, BBa_M0052, BBa_M0053). Sequence derived from reverse translation of AANDENYNYADAS sequence using codon usage optimized for E. coli. Three C-terminal aa's (DAS in this case) are necessary and sufficient for ClpX binding and degradation. Upstream aa sequence serves as a binding site for SspB, which guides rapid binding to ClpX. See sources on the main part page for further information about the mechanism of the system. NOTE: this sequence has only the amino acid sequence for the tag and bears NO STOP CODONS, so be sure to inlclude them if you use this sequence.
From a complete genomic sequence of Escherichia coli p0305293.5 gecp03052935.contig.229
The MioC Promoter is activated by the presence of DNAa which is produced in the S-Phase of bacterial replication. Isolated from Escherichia coli p0305293.5 gecp03052935.contig.229
Flpe is a site-specific DNA recombinase that that recognizes FRT sequence. Flpe was derived from yeaset site-specific DNA recombinase Flp. (Buchholz, et al.,Improved properties of FLP recombinase evolved by cycling mutagenesis. Nature. Biotechnology, 16:657-662 ( 1998 )) It's internal EcoRI and SpeI site has been removed using site-directed mutagenesis.
It's internal EcoRI and SpeI site has been removed using site-directed mutagenesis.
Kohaku So, Takashi Hiroi, Megumi Iseki, Mami Nakatake, Ryosuke Kamei, Ryo Kariyazono
Extend ca1010F/R (73 bp, EcoRI/SpeI) Sub into pSB1A2-I13522 (EcoRI/SpeI) Product is pSB1A2-Bca1010 ---- ca1010F Forward (universal biobrick) EcoRI for FRT GGACTgaattcgcggccgcttctagag ca1010R Reverse SpeI oligo for FRT cctatactagtagaagttcctattctctaAaaagtataggaacttcctctagaagcggccgcg
Site for recombination by flp recombinase. Genes flanked by FRT sites (oriented in the same direction) in the genome can be excised with the introduction of flp recombinase-expressing helper plasmid pCP20. Note that the original FRT sequence contained an XbaI site that has been removed with a point mutation for compatibility with standard assembly. See Datsenko and Wanner for details of its use in markerless knockouts and knockins.
The part accepts POPS to produce TetR. This sequence encodes the ribosome binding site and tetracycline repressor protein from the plasmid pASKIBA3.
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.
Lutz, R., Bujard, H., Nucleic Acids Research (1997) 25, 1203-1210.
Sequence for pTet inverting regulator driven by the TetR protein.
BBa_R0040 TetR-Regulated Promoter is based on a cI promoter. It has been modified to include two TetR binding sites and the BioBrick standard assembly head and tail restriction sites.
June Rhee, Connie Tao, Ty Thomson, Louis Waldman
λ -Hind Ⅲ digest (product of TaKaRa Bio)
This part causes cell death. It contains holin/antiholin, endolysin and Rz/Rz1 genes.
We made point mutation and prepared wild type phage DNA.
Conversion of the iGEM parts registry to SBOL2.1