This is an 'upside-down' version of I14032. It is a strong, constitual promoter. By adding this part, we wish to emphasise the possibility of engineering both DNA strands, and the additional reglatory possibility that this gives. The upstream sequence is non-functional and simply there to faciliate gel analysis or to physically enable the part to be recomineered (see the Edinburgh 2007 project where it is flanked by dif-sites).
When dealing with reverse complements of a locus, suffix and prefix need be fitted in opposite order.
This sequence is part of the E.coli K12 genome.
This part is the reverse of terminator B0013, a bacteriophage T7 terminator which was engineered to be bidirectional. Contains a poly A insertion upstream of a 20 bp stem-loop.
Reverse of bi-directional terminator B0051. (E.coli derived, genbank U24206 bases 2694-2727. between converging yciA and tonA genes.)
Inverted tetR promoter with noncoding, spacer DNA downstream of it to make the total length 250 bp. The promoter faces the 5' direction.
This part is in a series of inverted tetR promoters that have total lengths of 150, 250, 350, 450, 650, and 850 bp. We chose the region upstream of tetR in part Q04400 to be the noncoding DNA segment. This noncoding segment consists of the 3' end of tetR and B0015. None of the parts in this series was long enough to include the start codon of tetR, so a functional tetR protein should not be transcribed.
Primers were synthesized to bind to part Q04400, and then a PCR reaction was run.
Chloramphenicol resistance gene including its native promoter and ribosome binding site. Confers resistance to 25 ug/mL chloramphenicol. {pSB1A2-Bca1016}
PCR ca1016F/R on pKD3 (934 bp, EcoRI/SpeI) Sub into pSB1A2-I13522 (EcoRI/SpeI) Products are Bca1015 and Bca1016 ---- ca1016F Forward EcoRI to biobrick CAT of pKD3 gacttgaattcgcggccgcttctagaggaataggaacttcatttaaatg ca1016R Reverse SpeI to biobrick CAT of pKD3 ccgctactagtggcgcgcctacctgtgacgg
pT7-21 + RiboJ + GFP + LVA + 3 terminnators
This device is design for protein expression and promoter measurement. It constructed with a T7 RNA polymerase which has other 3 mutations, a RiboJ, a green fluorescent protein, a LVA degradation tag, a T7 terminator, a rrnB T1 terminator and a rrnB T2 terminator. The four T7 RNA polymerases are a family with different activity, the T7 RNA polymerase in this device have approximate relative activity 21%.
This device is design for protein expression and promoter measurement. It constructed with a T7 RNA polymerase which has other 3 mutations, a RiboJ, a green fluorescent protein, a LVA degradation tag, a T7 terminator, a rrnB T1 terminator and a rrnB T2 terminator. We added restriction sites flank these segments, so that users can change this segments into what they prefer. Users can replace T7 with other promoters through SalI and HindIII; replace RiboJ and GFP with other coding sequence through HindIII and StuI. After transcription, RiboJ becomes a section of functional RNA which comprises the sTRSV- ribozyme which can excise its upstream fragment to reduce noise, and an additional 23-nt hairpin immediately downstream to help expose the RBS. We use it to buffer synthetic circuits from genetic context. To adapt the protein expression device, three terminators are design to use cooperatively with strong promoters, the LVA degradation tag can accelerate protein degradation rate to decrease cell stress.
pT7-100 + RiboJ + GFP + LVA + 3 terminnators
This device is design for protein expression and promoter measurement. It constructed with a T7 RNA polymerase which has other 3 mutations, a RiboJ, a green fluorescent protein, a LVA degradation tag, a T7 terminator, a rrnB T1 terminator and a rrnB T2 terminator. The four T7 RNA polymerases are a family with different activity, the T7 RNA polymerase in this device have approximate relative activity 100%.
Reverse of B0053, E.coli transcriptional terminator from the His operon.
pT7-4 +RiboJ + GFP + LVA + 3 terminnators
This device is design for protein expression and promoter measurement. It constructed with a T7 RNA polymerase which has other 3 mutations, a RiboJ, a green fluorescent protein, a LVA degradation tag, a T7 terminator, a rrnB T1 terminator and a rrnB T2 terminator.
Inverted tetR promoter with noncoding, spacer DNA upstream of it to make the total length 450 bp.
pT7-10 + RiboJ + GFP + LVA + 3 terminnators
This device is design for protein expression and promoter measurement. It constructed with a T7 RNA polymerase which has other 3 mutations, a RiboJ, a green fluorescent protein, a LVA degradation tag, a T7 terminator, a rrnB T1 terminator and a rrnB T2 terminator. The four T7 RNA polymerases are a family with different activity, the T7 RNA polymerase in this device have approximate relative activity 10%.
tetR promoter with noncoding, spacer DNA upstream of it to make the total length 150 bp
This part is in a series of tetR promoter that have total lengths of 150, 250, 350, 450, 650, and 850 bp. We chose the region upstream of tetR in part Q04400 to be the noncoding DNA segment. This noncoding segment consists of the 3' end of tetR and B0015. None of the parts in this series was long enough to include the start codon of tetR, so a functional tetR protein should not be transcribed.
This is the same part as BBa_K110007 except it has Prefix and Suffix regions switched
This promoter is the right promoter in the lux cassette of Vibrio Fischeri. There is weak constitutive expression of downstream genes. Expression of downstream genes can be up-regulated by the action of the LuxR activator protein in complex with the autoinducer, 3-oxo-hexanoyl-HSL. Two molecules of LuxR protein can form a complex with two molecules of the AHL. This causes a conformational change in the protein, allowing it to bind to a site on the lux promoter and induce transcription.
This part was obtained by taking the reverse complement of R0062, ordering appropriate oligos, then annealing and extending them.
Reverse Bi-directional terminator from B0050 (derived from pBR322).
Inverted tetR promoter with noncoding, spacer DNA upstream of it to make the total length 650 bp.
Terminator from the E.coli ribosonal RNA rrnC operon. This terminator occurs in the reverse orientation upstream of the multiple cloning site of the pSB**1-pSB**3 series of plasmids.
Uni-directional terminator. Reverse of part B0012. 20 bp stem-loop.
Rick Carrick, Ambhi Ganesan, Alex McMillan
This is the same sequence as MFA1 promotor(BBa_K110008), but with suffix and prefix switched
Inverted tetR promoter with noncoding, spacer DNA upstream of it to make the total length 350 bp.
DNA is double-stranded. To translate genes of both strands, you need RBSs in both strands.
Please refer to the original (BB_B0034).
Transcriptional terminator consisting of a 64 bp stem-loop (B0010), cloned onto the antisense strand.
Antiquity. thought to be T1 transcriptional terminator from E. coli RRNB operon.
Reverse RBS (RBSrev) -- corresponds to BBa_B0030
Repeats in oligos caused unusual products during cloning