LOCUS       V015854                 8444 bp    DNA     circular SYN 01-APR-2020
DEFINITION  Exported.
ACCESSION   V015854
VERSION     V015854
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8444)
  AUTHORS   Shin J.
  TITLE     Programming Escherichia coli to function as a digital display
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8444)
  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 8444)
  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            "Unpublished"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..8444
                     /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..2166
                     /label="P2"
                     /note="P2"
                     /regulatory_class="ribosome_binding_site"
     CDS             2167..2769
                     /codon_start=1
                     /transl_table=11
                     /product="PhlF"
                     /label="PhlF"
                     /protein_id="QIQ56079.1"
                     /translation="MARTPSRSSIGSLRSPHTHKAILTSTIEILKECGYSGLSIESVAR
                     RAGASKPTIYRWWTNKAALIAEVYENESEQVRKFPDLGSFKADLDFLLRNLWKVWRETI
                     CGEAFRCVIAEAQLDPATLTQLKDQFMERRREMPKKLVENAISNGELPKDTNRELLLDM
                     IFGFCWYRLLTEQLTVEQDIEEFTFLLINGVCPGTQR"
     regulatory      2770..2827
                     /label="ECK120033737"
                     /note="ECK120033737"
                     /regulatory_class="terminator"
     protein_bind    2852..2870
                     /label="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    2906..2980
                     /label="RiboJ10"
                     /note="RiboJ10"
     regulatory      2981..3018
                     /label="S2"
                     /note="S2"
                     /regulatory_class="ribosome_binding_site"
     CDS             3019..3660
                     /codon_start=1
                     /transl_table=11
                     /product="SrpR"
                     /label="SrpR"
                     /protein_id="QIQ56080.1"
                     /translation="MARKTAAEAEETRQRIIDAALEVFVAQGVSDATLDQIARKAGVTR
                     GAVYWHFNGKLEVLQAVLASRQHPLELDFTPDLGIERSWEAVVVAMLDAVHSPQSKQFS
                     EILIYQGLDESGLIHNRMVQASDRFLQYIHQVLRHAVTQGELPINLDLQTSIGVFKGLI
                     TGLLYEGLRSKDQQAQIIKVALGSFWALLREPPRFLLCEEAQIKQVKSFE"
     regulatory      3661..3750
                     /label="ECK120029600"
                     /note="ECK120029600"
                     /regulatory_class="terminator"
     regulatory      3755..3820
                     /label="PPhlF"
                     /note="PPhlF"
                     /regulatory_class="promoter"
     misc_feature    3821..3895
                     /label="RiboJ51"
                     /note="RiboJ51"
     regulatory      3896..3904
                     /label="H1"
                     /note="H1"
                     /regulatory_class="ribosome_binding_site"
     CDS             3905..4540
                     /codon_start=1
                     /transl_table=11
                     /product="HlyIIR"
                     /label="HlyIIR"
                     /protein_id="QIQ56081.1"
                     /translation="MKYILFEVCEMGKSREQTMENILKAAKKKFGERGYEGTSIQEITK
                     EAKVNVAMASYYFNGKENLYYEVFKKYGLANELPNFLEKNQFNPINALREYLTVFTTHI
                     KENPEIGTLAYEEIIKESARLEKIKPYFIGSFEQLKEILQEGEKQGVFHFFSINHTIHW
                     ITSIVLFPKFKKFIDSLGPNETNDTNHEWMPEDLVSRIISALTDKPNI"
     regulatory      4541..4600
                     /label="ECK120010799"
                     /note="ECK120010799"
                     /regulatory_class="terminator"
     regulatory      4647..4700
                     /label="PLacI"
                     /note="PLacI"
                     /regulatory_class="promoter"
     regulatory      4701..4728
                     /label="a part of PLacI"
                     /note="a part of PLacI"
                     /regulatory_class="ribosome_binding_site"
     CDS             4729..5808
                     /label="lacI"
                     /note="lac repressor"
     regulatory      5812..5829
                     /regulatory_class="ribosome_binding_site"
     CDS             5830..6450
                     /label="TetR"
                     /note="tetracycline repressor TetR"
     regulatory      6454..6708
                     /regulatory_class="terminator"
     regulatory      6713..6747
                     /label="J23104"
                     /note="J23104"
                     /regulatory_class="promoter"
     regulatory      6748..6775
                     /regulatory_class="ribosome_binding_site"
     CDS             6776..7525
                     /gene="luxR"
                     /label="Transcriptional activator protein LuxR"
                     /note="Transcriptional activator protein LuxR from
                     Aliivibrio fischeri. Accession#: P12746"
     regulatory      7529..7556
                     /regulatory_class="ribosome_binding_site"
     CDS             7557..8312
                     /codon_start=1
                     /transl_table=11
                     /product="CinR"
                     /label="CinR"
                     /protein_id="QIQ56085.1"
                     /translation="MIENTYSEKFESAFEQIKAAANVDAAIRILQAEYNLDFVTYHLAQ
                     TIASKIDSPFVRTTYPDAWVSRYLLNCYVKVDPIIKQGFERQLPFDWSEVEPTPEAYAM
                     LVDAQKHGIDDNGYSIPVADKAQRRALLSLNAHIPADEWTELVRRCRNEWIEIAHLIHR
                     KAVYELHGENDPVPALSPREIECLHWTALGKDYKDISVILGISEHTTRDYLKTARFRLG
                     CTTISAAASRAVQLRIINPYRIRMTRRNW"
     terminator      8324..8395
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      8411..8438
                     /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 tttgtttaag gagctatgga ctatgtttga aaggctgaaa
     2161 tactagatgg cacgtacccc gagccgtagc agcattggta gcctgcgtag tccgcatacc
     2221 cataaagcaa ttctgaccag caccattgaa atcctgaaag aatgtggtta tagcggtctg
     2281 agcattgaaa gcgttgcacg tcgtgccggt gcaagcaaac cgaccattta tcgttggtgg
     2341 accaataaag cagcactgat tgccgaagtg tatgaaaatg aaagcgaaca ggtgcgtaaa
     2401 tttccggatc tgggtagctt taaagccgat ctggattttc tgctgcgtaa tctgtggaaa
     2461 gtttggcgtg aaaccatttg tggtgaagca tttcgttgtg ttattgcaga agcacagctg
     2521 gaccctgcaa ccctgaccca gctgaaagat cagtttatgg aacgtcgtcg tgagatgccg
     2581 aaaaaactgg ttgaaaatgc cattagcaat ggtgaactgc cgaaagatac caatcgtgaa
     2641 ctgctgctgg atatgatttt tggtttttgt tggtatcgcc tgctgaccga acagctgacc
     2701 gttgaacagg atattgaaga atttaccttc ctgctgatta atggtgtttg tccgggtaca
     2761 cagcgttaag gaaacacaga aaaaagcccg cacctgacag tgcgggcttt tttttttcga
     2821 ccaaaggtac gtactccacc gttggctttt ttccctatca gtgatagaga ttgacatccc
     2881 tatcagtgat agagataatg agcacagcgc tcaacgggtg tgcttcccgt tctgatgagt
     2941 ccgtgaggac gaaagcgcct ctacaaataa ttttgtttaa gagtctatgg actatgtttt
     3001 cacatatgag ataccaggat ggcacgtaaa accgcagcag aagcagaaga aacccgtcag
     3061 cgtattattg atgcagcact ggaagttttt gttgcacagg gtgttagtga tgcaaccctg
     3121 gatcagattg cacgtaaagc cggtgttacc cgtggtgcag tttattggca ttttaatggt
     3181 aaactggaag ttctgcaggc agttctggca agccgtcagc atccgctgga actggatttt
     3241 acaccggatc tgggtattga acgtagctgg gaagcagttg ttgttgcaat gctggatgca
     3301 gttcatagtc cgcagagcaa acagtttagc gaaattctga tttatcaggg tctggatgaa
     3361 agcggtctga ttcataatcg tatggttcag gcaagcgatc gttttctgca gtatattcat
     3421 caggttctgc gtcatgcagt tacccagggt gaactgccga ttaatctgga tctgcagacc
     3481 agcattggtg tttttaaagg tctgattacc ggtctgctgt atgaaggtct gcgtagcaaa
     3541 gatcagcagg cacagattat caaagttgca ctgggtagct tttgggcact gctgcgtgaa
     3601 ccgcctcgtt ttctgctgtg tgaagaagca cagattaaac aggtgaaatc cttcgaataa
     3661 ttcagccaaa aaacttaaga ccgccggtct tgtccactac cttgcagtaa tgcggtggac
     3721 aggatcggcg gttttctttt ctcttctcaa aggtcgacgt acggtggaat ctgattcgtt
     3781 accaattgac atgatacgaa acgtaccgta tcgttaaggt agtagtcacc ggctgtgctt
     3841 gccggtctga tgagcctgtg aaggcgaaac tacctctaca aataattttg tttaaacccc
     3901 cgagatgaaa tacatcctgt ttgaggtgtg cgaaatgggt aaaagccgtg aacagaccat
     3961 ggaaaatatt ctgaaagcag ccaaaaagaa attcggcgaa cgtggttatg aaggcaccag
     4021 cattcaagaa attaccaaag aagccaaagt taacgttgca atggccagct attactttaa
     4081 tggcaaagag aacctgtact acgaggtgtt caaaaaatac ggtctggcaa atgaactgcc
     4141 gaactttctg gaaaaaaacc agtttaatcc gattaatgcc ctgcgtgaat atctgaccgt
     4201 ttttaccacc cacattaaag aaaatccgga aattggcacc ctggcctatg aagaaattat
     4261 caaagaaagc gcacgcctgg aaaaaatcaa accgtatttt atcggcagct tcgaacagct
     4321 gaaagaaatt ctgcaagagg gtgaaaaaca gggtgtgttt cactttttta gcatcaacca
     4381 taccatccat tggattacca gcattgttct gtttccgaaa ttcaaaaaat tcatcgatag
     4441 cctgggtccg aatgaaacca atgataccaa tcatgaatgg atgccggaag atctggttag
     4501 ccgtattatt agcgcactga ccgataaacc gaacatttaa gttatgagtc aggaaaaaag
     4561 gcgacagagt aatctgtcgc cttttttctt tgcttgcttt aatgcgagac caattaaggc
     4621 ctgggaggcc agtttagaaa atggtagcgg cgcgccatcg aatggcgcaa aacctttcgc
     4681 ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatat gaaaccagta
     4741 acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg
     4801 aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag
     4861 ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt
     4921 ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa
     4981 tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc
     5041 gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt
     5101 aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg
     5161 gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgag
     5221 gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg
     5281 ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat
     5341 ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc
     5401 ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt
     5461 gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt
     5521 ggtgcggata tctcggtagt gggatacgac gataccgaag atagctcatg ttatatcccg
     5581 ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg
     5641 ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgccagt ctcactggtt
     5701 aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     5761 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg ataatccagg
     5821 aggaaaaaaa tgtccagatt agataaaagt aaagtgatta acagcgcatt agagctgctt
     5881 aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg cccagaagct aggtgtagag
     5941 cagcctacat tgtattggca tgtaaaaaat aagcgggctt tgctcgacgc cttagccatt
     6001 gagatgttag ataggcacca tactcacttt tgccctttag aaggggaaag ctggcaagat
     6061 tttttacgta ataacgctaa aagttttaga tgtgctttac taagtcatcg cgatggagca
     6121 aaagtacatt taggtacacg gcctacagaa aaacagtatg aaactctcga aaatcaatta
     6181 gcctttttat gccaacaagg tttttcacta gagaatgcat tatatgcact cagcgctgtg
     6241 gggcatttta ctttaggttg cgtattggaa gatcaagagc atcaagtcgc taaagaagaa
     6301 agggaaacac ctactactga tagtatgccg ccattattac gacaagctat cgaattattt
     6361 gatcaccaag gtgcagagcc agccttctta ttcggccttg aattgatcat atgcggatta
     6421 gaaaaacaac ttaaatgtga aagtgggtcc taataattgg taacgaatca gacaattgac
     6481 ggctcgaggg agtagcatag ggtttgcaga atccctgctt cgtccatttg acaggcacat
     6541 tatgcatcga tgataagctg tcaaacatga gcagatcctc tacgccggac gcatcgtggc
     6601 cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg
     6661 ggaagatcgg gctcgccact tcgggctcat gagcaaatat tttatctgag gtttgacagc
     6721 tagctcagtc ctaggtattg tgctagctct agagattaaa gaggagaaat actagatgaa
     6781 aaacataaat gccgacgaca catacagaat aattaataaa attaaagctt gtagaagcaa
     6841 taatgatatt aatcaatgct tatctgatat gactaaaatg gtacattgtg aatattattt
     6901 actcgcaatc atttatcctc attatatggt taaatccgat atttcaatcc tagataatta
     6961 ccctaaaaaa tggaggcaat attatgatga cgctaattta ataaaatatg atcctatagt
     7021 agattattct aactccaatc attcaccaat taattggaat atatttgaaa acaatgctgt
     7081 aaataaaaaa tctccaaatg taattaaaga agcgaaaaca tcaggtctta tcactgggtt
     7141 tagtttccct attcatacgg ctaacaatgg cttcggaatg cttagttttg cacattcaga
     7201 aaaagacaac tatatagata gtttattttt acatgcgtgt atgaacatac cattaattgt
     7261 tccttctcta gttgataatt atcgaaaaat aaatatagca aataataaat caaacaacga
     7321 tttaaccaaa agagaaaaag aatgtttagc gtgggcatgc gaaggaaaaa gctcttggga
     7381 tatttcaaaa atattaggtt gcagtgagcg tactgtcact ttccatttaa ccaatgcgca
     7441 aatgaaactc aatacaacaa accgctgcca aagtatttct aaagcaattt taacaggagc
     7501 aattgattgc ccatacttta aaaattaacc caaacaagta ccctcaggac gagataatga
     7561 ttgagaatac ctatagcgaa aagttcgagt ccgcgttcga acagatcaaa gcggcggcca
     7621 acgtggatgc cgccatccgt attctccagg cggaatataa cctcgatttc gtcacctacc
     7681 atctcgccca gacaatcgcg agcaagatcg attcgccctt cgtgcgcacc acctatccgg
     7741 atgcctgggt ttcccgttac ctcctcaact gctatgtgaa ggtcgatccg atcatcaagc
     7801 agggcttcga acgccagctg cccttcgact ggagcgaggt cgaaccgacg ccggaggcct
     7861 atgccatgct ggtcgacgcc cagaaacacg gcatcgatga caatggctac tccatccccg
     7921 tcgccgacaa ggcgcagcgc cgcgccctgc tgtcgctgaa tgcccatata ccggccgacg
     7981 aatggaccga gctcgtgcgc cgctgccgca atgagtggat cgagatcgcc catctgatcc
     8041 accgcaaggc cgtatatgag ctgcatggcg aaaacgatcc ggtgccggca ttgtcgccgc
     8101 gcgagatcga gtgtctgcac tggaccgccc tcggcaagga ttacaaggat atttcggtca
     8161 tcctgggcat atcagagcat accacacgcg attacctgaa aaccgcccgc ttcaggctcg
     8221 gctgcaccac gatctcggcc gccgcgtcgc gggctgttca attgcgcatc atcaatccct
     8281 ataggatccg catgacgcga cgtaattggt aatagccagg catcaaataa aacgaaaggc
     8341 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta
     8401 gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tata
//
