HRT2 prenyltransferase from Hevea Brasilianis (ara promoter with araC: arabinose inducible)
Apart from the prenyltransferase all the parts derive from E. coli. The prenyltransferase have been synthesized based on the cDNA sequence of a Hevea brasiliensis latex extract study. For more info, look at BBa_K1088003.
This biobrick expresses the Hevea brasiliensis rubber prenyltransferase when induced with IPTG.
There is a scarsite between the RBS of araC (BBa_B0030) aswell as in front of and behind the RBS of the prenyltransferase to improve translational efficieny. There are no more scarsites because the rest is assembled with USER cloning
AraC device (constitutive promoter, RBS and terminator)
>gnl|ECOLI|EG10525 lacI "PD00763" (complement(366734..365652)) Escherichia coli K-12 substr. MG1655
Coding region for the LacI protein derived from E. coli. LacI binds to the lac promoter BBa_R0010 and inhibits transcription. IPTG (Isopropyl β-D-1-thiogalactopyranoside) binds to LacI and inhibits its operation, thereby promoting transcription.
The GTG start codon was changed into ATG
HRT2 prenyltransferase from Hevea Brasilianis (ara promoter without araC: arabinose inducible)
This part encodes the rubberproducing prenyltransferase from the rubber tree Hevea brasiliensis. It uses isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) as substrates to produce cis-1,4 polyisoprene natural rubber. The gene is expressed from the arabinose promoter (BBa_I13453), which is induceable with arabinose under glucose scarce conditions. Note that the regulater insn't a part of this part and thus no/very little transcription is expected.
There is a 3xFLAG-tag after the HRT2 stop codon, and thus the 3xFLAG-tag isn't translated.
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.
strong synthetic E. coli ribosome binding site with SacI site.
This is a strong synthetic E. coli ribosome binding site. It is synthesised as two complementary oligonucleotides rather than being incorporated into a biobrick plasmid. It incorporates a SacI site overlapping the XbaI site, which is preserved when it is added to any other biobrick. This allows easy detection of the RBS after it has been added upstream of a biobrick coding sequence in a plasmid vector.
Note the presence of a SacI site overlapping the XbaI site, which is preserved when this biobrick is added to any other biobrick. At the time of writing, this biobrick is added as a short piece of DNA composed of two complementary oligonucleotides rather than being incorporated into a biobrick cloning vector by itself. It can be added upstream of a biobrick coding sequence, and its presence can easily be detected in miniprep DNA on a gel by using a SacI-SpeI or similar digest.
Bidirectional, with the reverse estimated to be less effective than the forward. Has a polyA tail of 6 residues.
Cis-1,4-prenyltransferase obtained from Hevea brasiliensis cDNA
The gene is synthesized by http://www.genscript.com/
This part encodes the rubberproducing prenyltransferase from the rubber tree Hevea brasiliensis. It uses isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) as substrates to produce cis-1,4 polyisoprene natural rubber. The sequence have been optimized for E. coli codons.
We optimized codons to fit the most used codons of E. coli.
Andreas Kj??r, Patrick Rosendahl Andreassen
arabinose regulator (araC) coding sequence
This sequence is a copy of the sequence 70,387 -> 71,265 in E. coli K-12 MG1655 which is the coding sequence of the pBAD repressor araC. From ECOCYC: The "arabinose regulator," AraC, is a transcription factor that regulates transcription of several genes and operons involved in arabinose catabolism and transport. It coregulates with another transcriptional regulator, CRP; both are transcription factors involved in l-arabinose degradation. These regulators bind cooperatively to activate transcription of five operons related to transport, catabolism, and autoregulation of l-arabinose. Transcription of these operons is induced when E. coli is grown in the absence of glucose and when the physiological inducer, l-arabinose, binds to the AraC regulator. In the absence of glucose, cellular cyclic AMP levels are high and cyclic AMP forms a dimeric complex with CRP to coregulate with AraC. For more info go to http://ecocyc.org/ECOLI/NEW-IMAGE?type=GENE&object=EG10054
The part was amplified from chromosomal DNA with primers carrying prefix and suffix overhangs
Mixed site (ACTAGA)when XbaI and SpeI sticky ends ligate
3xFLAG tag that can be fused with proteins either at the C- or N-terminal end. It can be used to purify the tagged protein or check levels of expression of tagged protein.
Conversion of the iGEM parts registry to SBOL2.1