Table 1.
Most burdensome BioBricks
| BioBricka | Seqb | Burden (b)c | Fraction other burden (bO/b)d | Subpartse | Functionf |
|---|---|---|---|---|---|
| K523022 | M | 51.7 ± 19.2% | n.s. | Plac &lacZ’ crtE crtI crtB | Carotenoid synthesis (Pantoea ananatis) |
| K733010 | C | 46.0 ± 6.2% | 0.16–0.71 | Ptms &endB | Antitoxin gene (Bacillus subtilis)B |
| J04450 | NS | 44.4 ± 2.2% | n.s. | Plac &mRFP1 | RFP reporter |
| K523014 | C | 39.6 ± 4.7% | 1.04–1.98 | Plac &lacZ’ bglX | Cellobiose degradation |
| K523020 | M, E | 38.3 ± 8.7% | n.s. | Plac &lacZ’ INP+bglX | Cellobiose degradation (INP, Pseudomonas syringae) |
| K608010 | C | 34.1 ± 7.7% | NT | PJ23110 &GFP | GFP reporter |
| K515100 | C | 33.9 ± 15.9% | 0.26–0.88 | Pveg2 &IaaM &IaaH | Indoleacetamide synthesis (Pseudomonas savastanoi)B |
| J61000 | m | 33.4 ± 4.1% | 0.21–0.96 | Pcat &cat | Chloramphenicol resistance |
| K541526 | C | 32.9 ± 7.8% | n.s. | Pveg &reflectin1A | Reflectin reporter (Euprymna scolopes)B |
| K592020 | m | 31.8 ± 5.0% | NT | PfixK2 &cI(λ) PcI &amilCP | Blue light sensor output (Acropora millepora) |
| J36335 | m | 30.2 ± 12.2% | n.s. | Plac &kaiA Plac &kaiC | Circadian rhythm (Synechococcus elongatus) |
| I759017 | C | 29.5 ± 8.3% | NT | Ptet [cis5] &YFP | YFP reporter |
| K346000 | C | 29.1 ± 10.4% | n.s. | &RNAP(T3) | Phage RNA polymerase (Phage T3) |
| C0056 | C | 28.2 ± 3.9% | n.s. | cI434(λ) | Mutant phage repressor (Phage λ) |
| K880005 | C | 27.5 ± 8.6% | n.s. | PJ23100 & | Gene expression |
| C0053 | NS | 27.2 ± 6.5% | n.s. | cII(P22) | Phage repressor (Phage P22) |
| K608012 | C | 27.1 ± 4.7% | NT | PJ23110 &GFP | GFP reporter |
| I759014 | C | 26.8 ± 5.8% | n.s. | Ptet [cis2] &YFP | YFP reporter |
| K541502 | C | 24.6 ± 3.2% | 0.42–1.91 | Pveg &lipAsig | Gene expression/secretion (Bacillus subtilis)B |
| K395602 | C | 20.3 ± 1.9% | 0.09–0.38 | PT7 &MpAAT1 | Apple fragrance generator (Malus pumila) |
| K733013 | C | 19.5 ± 3.3% | n.s. | Pveg &GFP | GFP reporterB |
| K523013 | C | 18.3 ± 8.8% | NT | Plac &lacZ’ INP + EYFP | EYFP reporter (INP, Pseudomonas syringae) |
| I761014 | C | 17.5 ± 5.0% | 0.21–1.33 | &cinR &cinI | Quorum sensing (Rhizobium leguminosarum) |
| C0051 | NS | 17.1 ± 8.2% | n.s. | λ-cI + LVA | Phage repressor (Phage λ) |
| K137018 | C | 16.8 ± 8.2% | NT | PL-lacO1 &luxR Plux-R &GFP | Quorum sensing receiver (Aliivibrio fischeri) |
| K1149051 | C | 15.0 ± 8.4% | n.s. | PJ23104 &phaC1 phaA phaB1 | Polyhydroxybutyrate synthesis (Ralstonia eutropha) |
| K731721 | C | 14.8 ± 4.4% | n.s. | Transcription terminator (Phage T7) | |
| K639003 | m | 14.8 ± 2.8% | n.s. | PrrnB-P1 &lacI PL-lacO1 &mCherry | Stress sensor |
| K541501 | C | 14.4 ± 3.6% | n.s. | Pveg &sacBsig | Gene expression/secretion (Bacillus subtilis)B |
| K608011 | C | 13.7 ± 5.4% | NT | PJ23110 &GFP | GFP reporter |
| K861172 | NS | 13.4 ± 2.5% | n.s. | PcstA &cI(λ) | Phage repressor (Phage λ) |
| K617004 | C | 11.6 ± 1.5% | 0.95–2.32 | attP(λ) P’OP | Phage attachment site (Phage λ) |
| K325218 | m | 10.8 ± 7.3% | 0.76–1.55 | ParaC &luc(orange) | Luciferase reporter (Luciola cruciata) |
| I712669 | m | 10.1 ± 4.5% | NT | PCMV GFP | GFP reporterM |
aBioBrick accession numbers. The 34 parts shown all had an estimated burden that was significantly greater than zero after correcting for multiple testing (one-tailed t-tests, Benjamini–Hochberg adjusted p < 0.05) and had a mean estimated burden value of >10%.
bResults of sequencing the BioBrick plasmid: C, reported BioBrick sequence was confirmed; M major discrepancies found in BioBrick sequence; m minor discrepancies found in BioBrick sequence; NS not sequenced; E part is reported to have errors in the iGEM Registry. Full sequencing results are provided in Supplementary Data 4.
cBurden as the percentage reduction in growth rate caused by the BioBrick ± estimated 95% confidence limits.
d95% confidence interval on the fraction of burden from sources other than utilization of the host cell’s gene expression capacity. n.s., value was not significantly greater than zero (one-tailed t-tests, Benjamini–Hochberg adjusted p < 0.05). NT not tested because the BioBrick contains a protein that interferes with the measurement of GFP fluorescence.
eRepresentation of gene expression signals and genes in the BioBrick abbreviated as follows: Px, promoter from gene or operon x; &, ribosome-binding site; [y] other regulatory sequence. Other italicized entries are gene names.
fGeneral description of the designed function of the BioBrick. For BioBricks that contain recombinant DNA encoding genes other than fluorescent proteins, the organism of origin is shown in parentheses. Superscript B or M, indicates that the gene expression sequences are intended to function in Bacillus subtilis or mammalian cells, respectively.