LOCUS Exported 8599 bp ds-DNA circular SYN 15-AUG-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..8599
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 26..70
/label=T5 promoter
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 46..62
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
protein_bind 78..94
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
RBS 104..115
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 122..1204
/codon_start=1
/gene="luxA"
/product="LuxA luciferase subunit"
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
KLFQSDVMPFLKEKQRSLLY"
RBS 1216..1227
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 1234..2217
/codon_start=1
/gene="luxB"
/product="LuxB luciferase subunit"
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MKFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENH
FSDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVRIAEEACLLDQLSEGRFILG
FSDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAY
TPGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEI
DHQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECIT
AAKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHMEYT"
RBS 2229..2240
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 2247..3689
/codon_start=1
/gene="luxC"
/product="LuxC fatty acid reductase"
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MTKKISFIINGQVEIFPESDDLVQSINFGDNSVYLPILNDSHVKN
IIDCNGNNELRLHNIVNFLYTVGQRWKNEEYSRRRTYIRDLKKYMGYSEEMAKLEANWI
SMILCSKGGLYDVVENELGSRHIMDEWLPQDESYVRAFPKGKSVHLLAGNVPLSGIMSI
LRAILTKNQCIIKTSSTDPFTANALALSFIDVDPNHPITRSLSVIYWPHQGDTSLAKEI
MRHADVIVAWGGPDAINWAVEHAPSYADVIKFGSKKSLCIIDNPVDLTSAATGAAHDVC
FYDQRACFSAQNIYYMGNHYEEFKLALIEKLNLYAHILPNAKKDFDEKAAYSLVQKESL
FAGLKVEVDIHQRWMIIESNAGVEFNQPLGRCVYLHHVDNIEQILPYVQKNKTQTISIF
PWESSFKYRDALALKGAERIVEAGMNNIFRVGGSHDGMRPLQRLVTYISHERPSNYTAK
DVAVEIEQTRFLEEDKFLVFVP"
RBS 3701..3712
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 3719..4642
/codon_start=1
/gene="luxD"
/product="LuxD acyltransferase"
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MENESKYKTIDHVICVEGNKKIHVWETLPEENSPKRKNAIIIASG
FARRMDHFAGLAEYLSRNGFHVIRYDSLHHVGLSSGTIDEFTMSIGKQSLLAVVDWLTT
NNFGMLASSLSARIAYASLSEINASFLITAVGVVNLRYSLERALGFDYLSLPINEL
PDNLDFEGHKLGAEVFARDCLDFGWEDLASTINNMMYLDIPFIAFTANNDNWVKQDEVI
TLLSNIRSNRCKIYSLLGSSHDLSENLVVLRNFYQSVTKAAIAMDNDHLDIDVDITEPS
FEHLTIATVNERRMRIEIENQAISLS"
RBS 4654..4665
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 4672..5784
/codon_start=1
/gene="luxE"
/product="LuxE"
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MTSYVDKQEITASSEIDDLIFSSDPLVWSYDEQEKIRKKLVLDAF
RNHYKHCREYRHYCQAHKVDDNITEIDDIPVFPTSVFKFTRLLTSQENEIESWFTSSGT
NGLKSQVARDRLSIERLLGSVSYGMKYVGSWFDHQIELVNLGPDRFNAHNIWFKYVMSL
VELLYPTTFTVTEERIDFVKTLNSLERIKNQGKDLCLIGSPYFIYLLCHYMKDKKISFS
GDKSLYIITGGGWKSYEKESLKRDDFNHLLFDTFNLSDISQIRDIFNQVELNTCFFEDE
MQRKHVPPWVYARALDPETLKPVPDGTPGLMSYMDASATSYPAFIVTDDVGIISREYGK
YPGVLVEILRRVNTRTQKGCALSLTEAFDS"
misc_feature 5788..5808
/label=BioBrick suffix
/note="universal suffix for all parts"
terminator 5859..5945
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 7489..7583
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(7599..8393)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEEYRCVKDTEGMSPAKVYKLVGENEILYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEGYED
EQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
1 aattcgcggc cgcttctaga ggaaatcata aaaaatttat ttgctttgtg agcggataac
61 aattataata gattcaattg tgagcggata acaattacta gagaaagagg agaaatacta
121 gatgaaattt ggaaactttt tgcttacata ccaacctccc caattttctc aaacagaggt
181 aatgaaacgt ttggttaaat taggtcgcat ctctgaggag tgtggttttg ataccgtatg
241 gttactggag catcatttca cggagtttgg tttgcttggt aacccttatg tcgctgctgc
301 atatttactt ggcgcgacta aaaaattgaa tgtaggaact gccgctattg ttcttcccac
361 agcccatcca gtacgccaac ttgaagatgt gaatttattg gatcaaatgt caaaaggacg
421 atttcggttt ggtatttgcc gagggcttta caacaaggac tttcgcgtat tcggcacaga
481 tatgaataac agtcgcgcct tagcggaatg ctggtacggg ctgataaaga atggcatgac
541 agagggatat atggaagctg ataatgaaca tatcaagttc cataaggtaa aagtaaaccc
601 cgcggcgtat agcagaggtg gcgcaccggt ttatgtggtg gctgaatcag cttcgacgac
661 tgagtgggct gctcaatttg gcctaccgat gatattaagt tggattataa atactaacga
721 aaagaaagca caacttgagc tttataatga agtggctcaa gaatatgggc acgatattca
781 taatatcgac cattgcttat catatataac atctgtagat catgactcaa ttaaagcgaa
841 agagatttgc cggaaatttc tggggcattg gtatgattct tatgtgaatg ctacgactat
901 ttttgatgat tcagaccaaa caagaggtta tgatttcaat aaagggcagt ggcgtgactt
961 tgtattaaaa ggacataaag atactaatcg ccgtattgat tacagttacg aaatcaatcc
1021 cgtgggaacg ccgcaggaat gtattgacat aattcaaaaa gacattgatg ctacaggaat
1081 atcaaatatt tgttgtggat ttgaagctaa tggaacagta gacgaaatta ttgcttccat
1141 gaagctcttc cagtctgatg tcatgccatt tcttaaagaa aaacaacgtt cgctattata
1201 ttaataatac tagagaaaga ggagaaatac tagatgaaat ttggattgtt cttccttaac
1261 ttcatcaatt caacaactgt tcaagaacaa agtatagttc gcatgcagga aataacggag
1321 tatgttgata agttgaattt tgaacagatt ttagtgtatg aaaatcattt ttcagataat
1381 ggtgttgtcg gcgctcctct gactgtttct ggttttctgc tcggtttaac agagaaaatt
1441 aaaattggtt cattaaatca catcattaca actcatcatc ctgtccgcat agcggaggaa
1501 gcttgcttat tggatcagtt aagtgaaggg agatttattt tagggtttag tgattgcgaa
1561 aaaaaagatg aaatgcattt ttttaatcgc ccggttgaat atcaacagca actatttgaa
1621 gagtgttatg aaatcattaa cgatgcttta acaacaggct attgtaatcc agataacgat
1681 ttttatagct tccctaaaat atctgtaaat ccccatgctt atacgccagg cggacctcgg
1741 aaatatgtaa cagcaaccag tcatcatatt gttgagtggg cggccaaaaa aggtattcct
1801 ctcatcttta agtgggatga ttctaatgat gttagatatg aatatgctga aagatataaa
1861 gccgttgcgg ataaatatga cgttgaccta tcagagatag accatcagtt aatgatatta
1921 gttaactata acgaagatag taataaagct aaacaagaga cgcgtgcatt tattagtgat
1981 tatgttcttg aaatgcaccc taatgaaaat ttcgaaaata aacttgaaga aataattgca
2041 gaaaacgctg tcggaaatta tacggagtgt ataactgcgg ctaagttggc aattgaaaag
2101 tgtggtgcga aaagtgtatt gctgtccttt gaaccaatga atgatttgat gagccaaaaa
2161 aatgtaatca atattgttga tgataatatt aagaagtacc acatggaata tacctaataa
2221 tactagagaa agaggagaaa tactagatga ctaaaaaaat ttcattcatt attaacggcc
2281 aggttgaaat ctttcccgaa agtgatgatt tagtgcaatc cattaatttt ggtgataata
2341 gtgtttacct gccaatattg aatgactctc atgtaaaaaa cattattgat tgtaatggaa
2401 ataacgaatt acggttgcat aacattgtca attttctcta tacggtaggg caaagatgga
2461 aaaatgaaga atactcaaga cgcaggacat acattcgtga cttaaaaaaa tatatgggat
2521 attcagaaga aatggctaag ctagaggcca attggatatc tatgatttta tgttctaaag
2581 gcggccttta tgatgttgta gaaaatgaac ttggttctcg ccatatcatg gatgaatggc
2641 tacctcagga tgaaagttat gttcgggctt ttccgaaagg taaatctgta catctgttgg
2701 caggtaatgt tccattatct gggatcatgt ctatattacg cgcaatttta actaagaatc
2761 agtgtattat aaaaacatcg tcaaccgatc cttttaccgc taatgcatta gcgttaagtt
2821 ttattgatgt agaccctaat catccgataa cgcgctcttt atctgttata tattggcccc
2881 accaaggtga tacatcactc gcaaaagaaa ttatgcgaca tgcggatgtt attgtcgctt
2941 ggggagggcc agatgcgatt aattgggcgg tagagcatgc gccatcttat gctgatgtga
3001 ttaaatttgg ttctaaaaag agtctttgca ttatcgataa tcctgttgat ttgacgtccg
3061 cagcgacagg tgcggctcat gatgtttgtt tttacgatca gcgagcttgt ttttctgccc
3121 aaaacatata ttacatggga aatcattatg aggaatttaa gttagcgttg atagaaaaac
3181 ttaatctata tgcgcatata ttaccgaatg ccaaaaaaga ttttgatgaa aaggcggcct
3241 attctttagt tcaaaaagaa agcttgtttg ctggattaaa agtagaggtg gatattcatc
3301 aacgttggat gattattgag tcaaatgcag gtgtggaatt taatcaacca cttggcagat
3361 gtgtgtacct tcatcacgtc gataatattg agcaaatatt gccttatgtt caaaaaaata
3421 agacgcaaac catatctatt tttccttggg agtcatcatt taaatatcga gatgcgttag
3481 cattaaaagg tgcggaaagg attgtagaag caggaatgaa taacatattt cgagttggtg
3541 gatctcatga cggaatgaga ccgttgcaac gattagtgac atatatttct catgaaaggc
3601 catctaacta tacggctaag gatgttgcgg ttgaaataga acagactcga ttcctggaag
3661 aagataagtt ccttgtattt gtcccataat aatactagag aaagaggaga aatactagat
3721 ggaaaatgaa tcaaaatata aaaccatcga ccacgttatt tgtgttgaag gaaataaaaa
3781 aattcatgtt tgggaaacgc tgccagaaga aaacagccca aagagaaaga atgccattat
3841 tattgcgtct ggttttgccc gcaggatgga tcattttgct ggtctggcgg aatatttatc
3901 gcggaatgga tttcatgtga tccgctatga ttcgcttcac cacgttggat tgagttcagg
3961 gacaattgat gaatttacaa tgtctatagg aaagcagagc ttgttagcag tggttgattg
4021 gttaactaca cgaaaaataa ataacttcgg tatgttggct tcaagcttat ctgcgcggat
4081 agcttatgca agcctatctg aaatcaatgc ttcgttttta atcaccgcag tcggtgttgt
4141 taacttaaga tattctcttg aaagagcttt agggtttgat tatctcagtc tacccattaa
4201 tgaattgccg gataatctgg attttgaagg ccataaattg ggtgctgaag tctttgcgag
4261 agattgtctt gattttggtt gggaagattt agcttctaca attaataaca tgatgtatct
4321 tgatataccg tttattgctt ttactgcaaa taacgataat tgggtcaagc aagatgaagt
4381 tatcacattg ttatcaaata ttcgtagtaa tcgatgcaag atatattctt tgttaggaag
4441 ttcgcatgac ttgagtgaaa atttagtggt cctgcgcaat ttttatcaat cggttacgaa
4501 agccgctatc gcgatggata atgatcatct ggatattgat gttgatatta ctgaaccgtc
4561 atttgaacat ttaactattg cgacagtcaa tgaacgccga atgagaattg agattgaaaa
4621 tcaagcaatt tctctgtctt aataatacta gagaaagagg agaaatacta gatgacttca
4681 tatgttgata aacaagaaat tacagcaagc tcagaaattg atgatttgat tttttcgagc
4741 gatccattag tgtggtctta cgacgagcag gaaaaaatca gaaagaaact tgtgcttgat
4801 gcatttcgta atcattataa acattgtcga gaatatcgtc actactgtca ggcacacaaa
4861 gtagatgaca atattacgga aattgatgac atacctgtat tcccaacatc ggtttttaag
4921 tttactcgct tattaacttc tcaggaaaac gagattgaaa gttggtttac cagtagcggc
4981 acgaatggtt taaaaagtca ggtggcgcgt gacagattaa gtattgagag actcttaggc
5041 tctgtgagtt atggcatgaa atatgttggt agttggtttg atcatcaaat agaattagtc
5101 aatttgggac cagatagatt taatgctcat aatatttggt ttaaatatgt tatgagtttg
5161 gtggaattgt tatatcctac gacatttacc gtaacagaag aacgaataga ttttgttaaa
5221 acattgaata gtcttgaacg aataaaaaat caagggaaag atctttgtct tattggttcg
5281 ccatacttta tttatttact ctgccattat atgaaagata aaaaaatctc attttctgga
5341 gataaaagcc tttatatcat aaccggaggc ggctggaaaa gttacgaaaa agaatctctg
5401 aaacgtgatg atttcaatca tcttttattt gatactttca atctcagtga tattagtcag
5461 atccgagata tatttaatca agttgaactc aacacttgtt tctttgagga tgaaatgcag
5521 cgtaaacatg ttccgccgtg ggtatatgcg cgagcgcttg atcctgaaac gttgaaacct
5581 gtacctgatg gaacgccggg gttgatgagt tatatggatg cgtcagcaac cagttatcca
5641 gcatttattg ttaccgatga tgtcgggata attagcagag aatatggtaa gtatcccggc
5701 gtgctcgttg aaattttacg tcgcgtcaat acgaggacgc agaaagggtg tgctttaagc
5761 ttaaccgaag cgtttgatag ttaataatac tagtagcggc cgctgcaggc ctcagggccc
5821 gatcgatgcc gccgcttaat taattaatcc agaggcatca aataaaacga aaggctcagt
5881 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
5941 caaatccgcc gccctagacc tagtgtcatt ttatttcccc cgtttcagca tcaagaacct
6001 ttgcataact tgctctatat ccacactgat aattgccctc aaaccataat ctaaaggcgc
6061 tagagtttgt tgaaacaata tcttttacat cattcgtatt taaaattcca aactccgctc
6121 ccctaaggcg aataaaagcc attaaatctt ttgtatttac caaattatag tcatccacta
6181 tatctaagag taaattcttc aattctcttt tttggctttc atcaagtgtt atatagcggt
6241 caatatcaaa atcattaatg ttcaaaatat cttttttgtc gtatatatgt ttattcttag
6301 caatagcgtc ctttgattca tgagtcaaat attcatatga acctttgata taatcaagta
6361 tctcaacatg agcaactgaa ctattcccca attttcgctt aatcttgttc ctaacgcttt
6421 ctattgttac aggatttcgt gcaatatata taacgtgata gtgtggtttt ttatagtgct
6481 ttccatttcg tataacatca ctactattcc atgtatcttt atcttttttt tcgtccatat
6541 cgtgtaaagg actgacagcc atagatacgc ccaaactctc taatttttcc ttccaatcat
6601 taggaattga gtcaggatat aataaaaatc caaaatttct agctttagta tttttaatag
6661 ccatgatata attaccttat caaaaacaag tagcgaaaac tcgtatcctt ctaaaaacgc
6721 gagctttcgc ttattttttt tgttctgatt cctttcttgc atattcttct atagctaacg
6781 ccgcaaccgc agattttgaa aaaccttttt gtttcgccat atctgttaat tttttatctt
6841 gctcttttgt cagagaaatc ataactcttt ttttcgattc tgaaatcacc atttaaaaaa
6901 ctccaatcaa ataattttat aaagttagtg tatcactttg taatcataaa aacaacaata
6961 aagctactta aatatagatt tataaaaaac gttggcgaaa acgttggcga ttcgttggcg
7021 attgaaaaac cccttaaacc cttgagccag ttgggataga gcgtttttgg cacaaaaatt
7081 ggcactcggc acttaatggg gggtcgtagt acggaagcaa aattcgcttc ctttcccccc
7141 atttttttcc aaattccaaa tttttttcaa aaattttcca gcgctaccgc tcggcaaaat
7201 tgcaagcaat ttttaaaatc aaacccatga gggaatttca ttccctcata ctcccttgag
7261 cctcctccaa ccgaaataga agggcgctgc gcttattatt tcattcagtc atcggctttc
7321 ataatctaac agacaacatc ttcgctgcaa agccacgcta cgctcaaggg cttttacgct
7381 acgataacgc ctgttttaac gattatgccg ataactaaac gaaataaacg ctaaaacgtc
7441 tcagaaacga ttttgagacg ttttaataaa aaatcgccta gtgcttggat tctcaccaat
7501 aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat
7561 tactggatct atcaacagga gtccaagcga gctcggtact aaaacaattc atccagtaaa
7621 atataatatt ttattttctc ccaatcaggc ttgatcccca gtaagtcaaa aaatagctcg
7681 acatactgtt cttccccgat atcctccctg atcgaccgga cgcagaaggc aatgtcatac
7741 cacttgtccg ccctgccgct tctcccaaga tcaataaagc cacttacttt gccatctttc
7801 acaaagatgt tgctgtctcc caggtcgccg tgggaaaaga caagttcctc ttcgggcttt
7861 tccgtcttta aaaaatcata cagctcgcgc ggatctttaa atggagtgtc ttcttcccag
7921 ttttcgcaat ccacatcggc cagatcgtta ttcagtaagt aatccaattc ggctaagcgg
7981 ccgtctaagc tattcgtata gggacaatcc gatatgtcga tggagtgaaa gagcctgatg
8041 cactccgcat acagctcgat agtcttttca gggctttgtt catcttcata cccttccgag
8101 caaaggacgc catcggcctc actcatgagc agattgctcc agccatcatg ccgttcaaag
8161 tgcaggacct ttggaacagg cagctttcct tccagccata gcatcatgtc cttttcccgt
8221 tccacatcat aggtggtccc tttataccgg ctgtccgtca tttttaaata taggatttca
8281 ttttctccca ccagcttata taccttagca ggagacattc cttccgtatc ttttacgcag
8341 cggtattctt cgatcagttt tttcaattcc ggtgatattc tcattttagc catttattat
8401 ttccttcctc ttttctacag tatttaaaga taccccaaga agctaattat aacaagacga
8461 actccaattc actgttcctt gcattctaaa accttaaata cagaaaacag ccttttcaaa
8521 gttgttttca aagttggcgt ataacatagt atcgacggag ccgattttga aaccacaatt
8581 atgatagaat ttgacgtcg
//