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

                     RKINNFGMLASSLSARIAYASLSEINASFLITAVGVVNLRYSLERALGFDYLSLPINEL

                     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

//

Caution:
Product is for research use only!