LOCUS       V015492                 8644 bp    DNA     circular SYN 01-APR-2020
DEFINITION  Exported.
ACCESSION   V015492
VERSION     V015492
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8644)
  AUTHORS   Shin J.
  TITLE     Programming Escherichia coli to function as a digital display
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8644)
  AUTHORS   Shin J.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-JAN-2020) Biomedical Engineering, Boston University,
            44 Cummington Mall, Boston, MA 02215, USA
REFERENCE   3  (bases 1 to 8644)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (19-JAN-2020) Biomedical Engineering, Boston University, 44
            Cummington Mall, Boston, MA 02215, USA"
FEATURES             Location/Qualifiers
     source          1..8644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(297..842)
                     /direction=LEFT
                     /label="p15A ori"
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells
                     that contain a second plasmid with the ColE1 origin."
     CDS             complement(934..1746)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     regulatory      1903..1984
                     /label="PSrpR"
                     /note="PSrpR"
                     /regulatory_class="promoter"
     regulatory      1985..2054
                     /label="PLux2"
                     /note="PLux2"
                     /regulatory_class="promoter"
     misc_feature    2055..2129
                     /label="RiboJ53"
                     /note="RiboJ53"
     regulatory      2130..2147
                     /label="P3"
                     /note="P3"
                     /regulatory_class="ribosome_binding_site"
     CDS             2148..2750
                     /codon_start=1
                     /transl_table=11
                     /product="PhlF"
                     /label="PhlF"
                     /protein_id="QIQ56099.1"
                     /translation="MARTPSRSSIGSLRSPHTHKAILTSTIEILKECGYSGLSIESVAR
                     RAGASKPTIYRWWTNKAALIAEVYENESEQVRKFPDLGSFKADLDFLLRNLWKVWRETI
                     CGEAFRCVIAEAQLDPATLTQLKDQFMERRREMPKKLVENAISNGELPKDTNRELLLDM
                     IFGFCWYRLLTEQLTVEQDIEEFTFLLINGVCPGTQR"
     regulatory      2751..2808
                     /label="ECK120033737"
                     /note="ECK120033737"
                     /regulatory_class="terminator"
     regulatory      2813..2875
                     /label="PHlyIIR2"
                     /note="PHlyIIR2"
                     /regulatory_class="promoter"
     protein_bind    2896..2914
                     /label="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    2950..3024
                     /label="RiboJ10"
                     /note="RiboJ10"
     regulatory      3025..3058
                     /label="S4"
                     /note="S4"
                     /regulatory_class="ribosome_binding_site"
     CDS             3059..3700
                     /codon_start=1
                     /transl_table=11
                     /product="SrpR"
                     /label="SrpR"
                     /protein_id="QIQ56100.1"
                     /translation="MARKTAAEAEETRQRIIDAALEVFVAQGVSDATLDQIARKAGVTR
                     GAVYWHFNGKLEVLQAVLASRQHPLELDFTPDLGIERSWEAVVVAMLDAVHSPQSKQFS
                     EILIYQGLDESGLIHNRMVQASDRFLQYIHQVLRHAVTQGELPINLDLQTSIGVFKGLI
                     TGLLYEGLRSKDQQAQIIKVALGSFWALLREPPRFLLCEEAQIKQVKSFE"
     regulatory      3701..3790
                     /label="ECK120029600"
                     /note="ECK120029600"
                     /regulatory_class="terminator"
     regulatory      3795..4020
                     /label="PCin"
                     /note="PCin"
                     /regulatory_class="promoter"
     misc_feature    4021..4095
                     /label="RiboJ51"
                     /note="RiboJ51"
     regulatory      4096..4104
                     /label="H1"
                     /note="H1"
                     /regulatory_class="ribosome_binding_site"
     CDS             4105..4740
                     /codon_start=1
                     /transl_table=11
                     /product="HlyIIR"
                     /label="HlyIIR"
                     /protein_id="QIQ56101.1"
                     /translation="MKYILFEVCEMGKSREQTMENILKAAKKKFGERGYEGTSIQEITK
                     EAKVNVAMASYYFNGKENLYYEVFKKYGLANELPNFLEKNQFNPINALREYLTVFTTHI
                     KENPEIGTLAYEEIIKESARLEKIKPYFIGSFEQLKEILQEGEKQGVFHFFSINHTIHW
                     ITSIVLFPKFKKFIDSLGPNETNDTNHEWMPEDLVSRIISALTDKPNI"
     regulatory      4741..4800
                     /label="ECK120010799"
                     /note="ECK120010799"
                     /regulatory_class="terminator"
     regulatory      4847..4900
                     /label="PLacI"
                     /note="PLacI"
                     /regulatory_class="promoter"
     regulatory      4901..4928
                     /label="a part of PLacI"
                     /note="a part of PLacI"
                     /regulatory_class="ribosome_binding_site"
     CDS             4929..6008
                     /label="lacI"
                     /note="lac repressor"
     regulatory      6012..6029
                     /regulatory_class="ribosome_binding_site"
     CDS             6030..6650
                     /label="TetR"
                     /note="tetracycline repressor TetR"
     regulatory      6654..6908
                     /regulatory_class="terminator"
     regulatory      6948..6975
                     /regulatory_class="ribosome_binding_site"
     CDS             6976..7725
                     /gene="luxR"
                     /label="Transcriptional activator protein LuxR"
                     /note="Transcriptional activator protein LuxR from
                     Aliivibrio fischeri. Accession#: P12746"
     regulatory      7729..7756
                     /regulatory_class="ribosome_binding_site"
     CDS             7757..8512
                     /codon_start=1
                     /transl_table=11
                     /product="CinR"
                     /label="CinR"
                     /protein_id="QIQ56105.1"
                     /translation="MIENTYSEKFESAFEQIKAAANVDAAIRILQAEYNLDFVTYHLAQ
                     TIASKIDSPFVRTTYPDAWVSRYLLNCYVKVDPIIKQGFERQLPFDWSEVEPTPEAYAM
                     LVDAQKHGIDDNGYSIPVADKAQRRALLSLNAHIPADEWTELVRRCRNEWIEIAHLIHR
                     KAVYELHGENDPVPALSPREIECLHWTALGKDYKDISVILGISEHTTRDYLKTARFRLG
                     CTTISAAASRAVQLRIINPYRIRMTRRNW"
     terminator      8524..8595
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      8611..8638
                     /label="T7Te terminator"
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 cactgactcg ctgcacgagg cagacctcag cgctagcgga gtgtatactg gcttactatg
       61 ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc
      121 ggtgcgtcag cagaatatgt gatacaggat atattccgct tcctcgctca ctgactcgct
      181 acgctcggtc gttcgactgc ggcgagcgga aatggcttac gaacggggcg gagatttcct
      241 ggaagatgcc aggaagatac ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc
      301 cataggctcc gcccccctga caagcatcac gaaatctgac gctcaaatca gtggtggcga
      361 aacccgacag gactataaag ataccaggcg tttccccctg gcggctccct cgtgcgctct
      421 cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg tttgtctcat
      481 tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact gtatgcacga
      541 accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
      601 ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta gaggagttag
      661 tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt gactgcgctc
      721 ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc
      781 cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca aaacgatctc
      841 aagaagatca tcttattaag gggtctgacg ctcagtggaa cgaaaaatca atctaaagta
      901 tatatgagta aacttggtct gacagttacc ttagaaaaac tcatcgagca tcaaatgaaa
      961 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     1021 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     1081 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     1141 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg
     1201 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     1261 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct
     1321 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     1381 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc
     1441 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     1501 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     1561 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     1621 tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     1681 atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc gttgaatatg
     1741 gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga
     1801 tatattttta tcttgtgcaa tgtacatcag agattttgag acacaaagcc tgcctgtcta
     1861 ttgatgcggc cgaaacggcc agattgtact gggtctctgg agtctatgat tggtccagat
     1921 tcgttaccaa ttgacagcta gctcagtcct aggtatatac atacatgctt gtttgtttgt
     1981 aaacatagct tcttaccgga cctgtaggat cgtacaggtt tacgcaagaa aatggtttgt
     2041 tattttcgaa taaaagcggt caacgcatgt gctttgcgtt ctgatgagac agtgatgtcg
     2101 aaaccgcctc tacaaataat tttgtttaac tttacgaggg cgatcctatg gcacgtaccc
     2161 cgagccgtag cagcattggt agcctgcgta gtccgcatac ccataaagca attctgacca
     2221 gcaccattga aatcctgaaa gaatgtggtt atagcggtct gagcattgaa agcgttgcac
     2281 gtcgtgccgg tgcaagcaaa ccgaccattt atcgttggtg gaccaataaa gcagcactga
     2341 ttgccgaagt gtatgaaaat gaaagcgaac aggtgcgtaa atttccggat ctgggtagct
     2401 ttaaagccga tctggatttt ctgctgcgta atctgtggaa agtttggcgt gaaaccattt
     2461 gtggtgaagc atttcgttgt gttattgcag aagcacagct ggaccctgca accctgaccc
     2521 agctgaaaga tcagtttatg gaacgtcgtc gtgagatgcc gaaaaaactg gttgaaaatg
     2581 ccattagcaa tggtgaactg ccgaaagata ccaatcgtga actgctgctg gatatgattt
     2641 ttggtttttg ttggtatcgc ctgctgaccg aacagctgac cgttgaacag gatattgaag
     2701 aatttacctt cctgctgatt aatggtgttt gtccgggtac acagcgttaa ggaaacacag
     2761 aaaaaagccc gcacctgaca gtgcgggctt ttttttttcg accaaaggta cgaccaggaa
     2821 tctgaacgat tcgttaccaa ttgccatatt taaaattctt gtttaaaatg ctagctactc
     2881 caccgttggc ttttttccct atcagtgata gagattgaca tccctatcag tgatagagat
     2941 aatgagcaca gcgctcaacg ggtgtgcttc ccgttctgat gagtccgtga ggacgaaagc
     3001 gcctctacaa ataattttgt ttaactatgg actatgtttt cacacaggaa ataccaggat
     3061 ggcacgtaaa accgcagcag aagcagaaga aacccgtcag cgtattattg atgcagcact
     3121 ggaagttttt gttgcacagg gtgttagtga tgcaaccctg gatcagattg cacgtaaagc
     3181 cggtgttacc cgtggtgcag tttattggca ttttaatggt aaactggaag ttctgcaggc
     3241 agttctggca agccgtcagc atccgctgga actggatttt acaccggatc tgggtattga
     3301 acgtagctgg gaagcagttg ttgttgcaat gctggatgca gttcatagtc cgcagagcaa
     3361 acagtttagc gaaattctga tttatcaggg tctggatgaa agcggtctga ttcataatcg
     3421 tatggttcag gcaagcgatc gttttctgca gtatattcat caggttctgc gtcatgcagt
     3481 tacccagggt gaactgccga ttaatctgga tctgcagacc agcattggtg tttttaaagg
     3541 tctgattacc ggtctgctgt atgaaggtct gcgtagcaaa gatcagcagg cacagattat
     3601 caaagttgca ctgggtagct tttgggcact gctgcgtgaa ccgcctcgtt ttctgctgtg
     3661 tgaagaagca cagattaaac aggtgaaatc cttcgaataa ttcagccaaa aaacttaaga
     3721 ccgccggtct tgtccactac cttgcagtaa tgcggtggac aggatcggcg gttttctttt
     3781 ctcttctcaa aggtcccttt gtgcgtccaa acggacgcac ggcgctctaa agcgggtcgc
     3841 gatctttcag attcgctcct cgcgctttca gtctttgttt tggcgcatgt cgttatcgca
     3901 aaaccgctgc acacttttgc gcgacatgct ctgatccccc tcatctgggg gggcctatct
     3961 gagggaattt ccgatccggc tcgcctgaac cattctgctt tccacgaact tgaaaacgct
     4021 agtagtcacc ggctgtgctt gccggtctga tgagcctgtg aaggcgaaac tacctctaca
     4081 aataattttg tttaaacccc cgagatgaaa tacatcctgt ttgaggtgtg cgaaatgggt
     4141 aaaagccgtg aacagaccat ggaaaatatt ctgaaagcag ccaaaaagaa attcggcgaa
     4201 cgtggttatg aaggcaccag cattcaagaa attaccaaag aagccaaagt taacgttgca
     4261 atggccagct attactttaa tggcaaagag aacctgtact acgaggtgtt caaaaaatac
     4321 ggtctggcaa atgaactgcc gaactttctg gaaaaaaacc agtttaatcc gattaatgcc
     4381 ctgcgtgaat atctgaccgt ttttaccacc cacattaaag aaaatccgga aattggcacc
     4441 ctggcctatg aagaaattat caaagaaagc gcacgcctgg aaaaaatcaa accgtatttt
     4501 atcggcagct tcgaacagct gaaagaaatt ctgcaagagg gtgaaaaaca gggtgtgttt
     4561 cactttttta gcatcaacca taccatccat tggattacca gcattgttct gtttccgaaa
     4621 ttcaaaaaat tcatcgatag cctgggtccg aatgaaacca atgataccaa tcatgaatgg
     4681 atgccggaag atctggttag ccgtattatt agcgcactga ccgataaacc gaacatttaa
     4741 gttatgagtc aggaaaaaag gcgacagagt aatctgtcgc cttttttctt tgcttgcttt
     4801 aatgcgagac caattaaggc ctgggaggcc agtttagaaa atggtagcgg cgcgccatcg
     4861 aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg
     4921 tggtgaatat gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc
     4981 agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag
     5041 tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg
     5101 gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc
     5161 aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga
     5221 tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac
     5281 gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag
     5341 ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca
     5401 gtattatttt ctcccatgag gacggtacgc gactgggcgt ggagcatctg gtcgcattgg
     5461 gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc
     5521 tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag
     5581 gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg
     5641 ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta
     5701 ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag
     5761 atagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc
     5821 aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc
     5881 tgttgccagt ctcactggtt aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct
     5941 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     6001 gcgggcagtg ataatccagg aggaaaaaaa tgtccagatt agataaaagt aaagtgatta
     6061 acagcgcatt agagctgctt aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg
     6121 cccagaagct aggtgtagag cagcctacat tgtattggca tgtaaaaaat aagcgggctt
     6181 tgctcgacgc cttagccatt gagatgttag ataggcacca tactcacttt tgccctttag
     6241 aaggggaaag ctggcaagat tttttacgta ataacgctaa aagttttaga tgtgctttac
     6301 taagtcatcg cgatggagca aaagtacatt taggtacacg gcctacagaa aaacagtatg
     6361 aaactctcga aaatcaatta gcctttttat gccaacaagg tttttcacta gagaatgcat
     6421 tatatgcact cagcgctgtg gggcatttta ctttaggttg cgtattggaa gatcaagagc
     6481 atcaagtcgc taaagaagaa agggaaacac ctactactga tagtatgccg ccattattac
     6541 gacaagctat cgaattattt gatcaccaag gtgcagagcc agccttctta ttcggccttg
     6601 aattgatcat atgcggatta gaaaaacaac ttaaatgtga aagtgggtcc taataattgg
     6661 taacgaatca gacaattgac ggctcgaggg agtagcatag ggtttgcaga atccctgctt
     6721 cgtccatttg acaggcacat tatgcatcga tgataagctg tcaaacatga gcagatcctc
     6781 tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat
     6841 atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcaaatat
     6901 tttatctgag gtttgacagc tagctcagtc ctaggtattg tgctagctct agagattaaa
     6961 gaggagaaat actagatgaa aaacataaat gccgacgaca catacagaat aattaataaa
     7021 attaaagctt gtagaagcaa taatgatatt aatcaatgct tatctgatat gactaaaatg
     7081 gtacattgtg aatattattt actcgcaatc atttatcctc attatatggt taaatccgat
     7141 atttcaatcc tagataatta ccctaaaaaa tggaggcaat attatgatga cgctaattta
     7201 ataaaatatg atcctatagt agattattct aactccaatc attcaccaat taattggaat
     7261 atatttgaaa acaatgctgt aaataaaaaa tctccaaatg taattaaaga agcgaaaaca
     7321 tcaggtctta tcactgggtt tagtttccct attcatacgg ctaacaatgg cttcggaatg
     7381 cttagttttg cacattcaga aaaagacaac tatatagata gtttattttt acatgcgtgt
     7441 atgaacatac cattaattgt tccttctcta gttgataatt atcgaaaaat aaatatagca
     7501 aataataaat caaacaacga tttaaccaaa agagaaaaag aatgtttagc gtgggcatgc
     7561 gaaggaaaaa gctcttggga tatttcaaaa atattaggtt gcagtgagcg tactgtcact
     7621 ttccatttaa ccaatgcgca aatgaaactc aatacaacaa accgctgcca aagtatttct
     7681 aaagcaattt taacaggagc aattgattgc ccatacttta aaaattaacc caaacaagta
     7741 ccctcaggac gagataatga ttgagaatac ctatagcgaa aagttcgagt ccgcgttcga
     7801 acagatcaaa gcggcggcca acgtggatgc cgccatccgt attctccagg cggaatataa
     7861 cctcgatttc gtcacctacc atctcgccca gacaatcgcg agcaagatcg attcgccctt
     7921 cgtgcgcacc acctatccgg atgcctgggt ttcccgttac ctcctcaact gctatgtgaa
     7981 ggtcgatccg atcatcaagc agggcttcga acgccagctg cccttcgact ggagcgaggt
     8041 cgaaccgacg ccggaggcct atgccatgct ggtcgacgcc cagaaacacg gcatcgatga
     8101 caatggctac tccatccccg tcgccgacaa ggcgcagcgc cgcgccctgc tgtcgctgaa
     8161 tgcccatata ccggccgacg aatggaccga gctcgtgcgc cgctgccgca atgagtggat
     8221 cgagatcgcc catctgatcc accgcaaggc cgtatatgag ctgcatggcg aaaacgatcc
     8281 ggtgccggca ttgtcgccgc gcgagatcga gtgtctgcac tggaccgccc tcggcaagga
     8341 ttacaaggat atttcggtca tcctgggcat atcagagcat accacacgcg attacctgaa
     8401 aaccgcccgc ttcaggctcg gctgcaccac gatctcggcc gccgcgtcgc gggctgttca
     8461 attgcgcatc atcaatccct ataggatccg catgacgcga cgtaattggt aatagccagg
     8521 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     8581 tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt
     8641 tata
//
