LOCUS       V002649                 7391 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002649
VERSION     V002649
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7391)
  AUTHORS   Segall-Shapiro TH, Sontag ED, Voigt CA.
  TITLE     Engineered promoters enable constant gene expression at any copy
            number in bacteria
  JOURNAL   Nat. Biotechnol. (2018) In press
   PUBMED   29553576
REFERENCE   2  (bases 1 to 7391)
  AUTHORS   Segall-Shapiro TH, Sontag ED, Voigt CA.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-MAR-2018) Synthetic Biology Center, Department of
            Biological Engineering, Massachusetts Institute of Technology, 77
            Massachusetts Avenue, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 7391)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7391)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat.
            Biotechnol. (2018) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (06-MAR-2018) Synthetic Biology Center, Department of Biological
            Engineering, Massachusetts Institute of Technology, 77 Massachusetts
            Avenue, Cambridge, MA 02139, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      complement(13..102)
                     /label="ECK120029600"
                     /note="ECK120029600"
                     /regulatory_class="terminator"
     regulatory      107..175
                     /label="PT7A1w1"
                     /note="PT7A1w1"
                     /regulatory_class="promoter"
     misc_RNA        177..227
                     /label="sTRSV HHRz"
                     /note="hammerhead ribozyme from the tobacco ringspot virus
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      251..273
                     /label="RBSVioEc"
                     /note="RBSVioEc"
                     /regulatory_class="ribosome_binding_site"
     CDS             274..849
                     /codon_start=1
                     /product="VioE"
                     /label="VioE"
                     /protein_id="AVR55197.1"
                     /translation="MENREPPLLPARWSSAYVSYWSPMLPDDQLTSGYCWFDYERDICR
                     IDGLFNPWSERDTGYRLWMSEVGNAASGRTWKQKVAYGRERTALGEQLCERPLDDETGP
                     FAELFLPRDVLRRLGARHIGRRVVLGREADGWRYQRPGKGPSTLYLDAASGTPLRMVTG
                     DEASRASLRDFPNVSEAEIPDAVFAAKR"
     regulatory      850..873
                     /label="RBSVioAc"
                     /note="RBSVioAc"
                     /regulatory_class="ribosome_binding_site"
     CDS             874..2127
                     /gene="vioA"
                     /label="Flavin-dependent L-tryptophan oxidase VioA"
                     /note="Flavin-dependent L-tryptophan oxidase VioA from
                     Chromobacterium violaceum (strain ATCC 12472 / DSM 30191 /
                     JCM 1249 / NBRC 12614 / NCIMB 9131 / NCTC 9757).
                     Accession#: Q9S3V1"
     regulatory      2131..2154
                     /label="RBSVioBc"
                     /note="RBSVioBc"
                     /regulatory_class="ribosome_binding_site"
     CDS             2155..5148
                     /gene="vioB"
                     /label="2-imino-3-(indol-3-yl)propanoate dimerase"
                     /note="2-imino-3-(indol-3-yl)propanoate dimerase from
                     Chromobacterium violaceum (strain ATCC 12472 / DSM 30191 /
                     JCM 1249 / NBRC 12614 / NCIMB 9131 / NCTC 9757).
                     Accession#: Q9S3V0"
     terminator      5166..5237
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      5253..5280
                     /label="T7Te terminator"
                     /note="phage T7 early transcription terminator"
     misc_feature    5299..5319
                     /label="BioBrick suffix"
                     /note="universal suffix for all parts"
     terminator      5320..5377
                     /label="his operon terminator"
                     /note="This putative transcriptin terminator from the E.
                     coli his operon has a 2-bp deletion introduced during
                     synthesis. Its efficiency has not been determined."
     rep_origin      5549..6137
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6338..7129)
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     terminator      complement(7324..7367)
                     /label="bacterial terminator"
                     /note="putative bacterial transcription terminator"
     misc_feature    7370..7391
                     /label="BioBrick prefix"
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
ORIGIN
        1 gaagacacgg agttcagcca aaaaacttaa gaccgccggt cttgtccact accttgcagt
       61 aatgcggtgg acaggatcgg cggttttctt ttctcttctc aagcttggcg cgcctcagtc
      121 agagtattga cttaaagtct aacctatagg acctttacag ccatcgagag ctgcgagctg
      181 tcaccggatg tgctttccgg tctgatgagt ccgtgaggac gaaacagcct ctacaaataa
      241 ttttgtttaa tactcattag ggcgggaaaa aaaatggaga accgtgagcc accactgttg
      301 ccagcccgtt ggagcagcgc ctatgtctct tattggagcc cgatgctgcc ggatgaccag
      361 ctgaccagcg gctattgctg gttcgactat gaacgtgaca tctgtcgtat tgacggcctg
      421 ttcaatccgt ggagcgagcg tgatactggt tatcgcctgt ggatgtcgga ggttggtaat
      481 gcggccagcg gccgtacctg gaaacaaaaa gtcgcctatg gtcgtgagcg taccgccctg
      541 ggtgaacagc tgtgtgagcg tccgctggat gatgagactg gcccttttgc cgaattgttc
      601 ctgccacgcg atgtcctgcg ccgtctgggt gcccgtcaca ttggccgtcg cgtggttctg
      661 ggtcgcgaag cggacggttg gcgttaccag cgcccaggta aaggtccgag caccctgtac
      721 ctggatgcgg cgagcggcac tccactgcgc atggtcaccg gcgatgaagc gtcgcgtgca
      781 agcctgcgtg attttccgaa tgtgagcgag gcggagatcc cggacgcggt tttcgcggcc
      841 aagcgctaat aacattatat ggcggaaaaa aaaatgaaac attcttccga tatctgcatt
      901 gttggtgctg gtatttctgg tttgacgtgc gcaagccatc tgctggacag cccggcatgc
      961 cgtggtctga gcctgcgtat ctttgacatg cagcaagaag ccggtggccg tatccgcagc
     1021 aaaatgctgg atggtaaggc aagcattgaa ctgggcgcag gtcgctactc ccctcagttg
     1081 cacccgcatt tccaaagcgc aatgcagcac tatagccaaa agagcgaagt ctatccgttc
     1141 acccagttga agttcaaatc tcacgtgcag caaaagctga agcgcgccat gaatgaactg
     1201 tccccgcgtc tgaaagagca tggtaaagag agctttttgc agtttgtcag ccgttatcaa
     1261 ggtcacgata gcgcggttgg tatgatccgc tctatgggtt acgacgcact gttcctgccg
     1321 gatatcagcg cagaaatggc ctacgacatt gtgggtaagc acccggagat ccagagcgtg
     1381 acggacaacg acgcgaacca atggtttgca gcggaaacgg gctttgctgg tctgattcag
     1441 ggcatcaagg ctaaggttaa ggcggcaggt gcgcgtttta gcctgggtta tcgtctgctg
     1501 agcgtccgta ccgacggtga cggctacctg ctgcaactgg caggtgacga cggctggaaa
     1561 ctggagcacc gtacccgcca tctgattctg gcgattccgc cgagcgcgat ggcgggtttg
     1621 aatgttgatt ttccagaagc ctggtccggt gcgcgctatg gcagcctgcc gctgtttaag
     1681 ggctttctga cgtacggtga gccgtggtgg ttggactaca aactggacga tcaggtgctg
     1741 attgttgaca acccgctgcg caaaatctat ttcaaaggcg ataagtacct gttcttctat
     1801 accgatagcg agatggcgaa ttactggcgc ggttgtgtcg cggagggcga ggacggttac
     1861 ctggagcaaa ttcgcaccca tttggctagc gcactgggta tcgtccgtga acgtatcccg
     1921 caaccgctgg cacacgttca caagtattgg gcgcacggcg ttgagttttg ccgtgattct
     1981 gatattgacc acccgagcgc actgtctcat cgcgacagcg gtatcatcgc gtgctccgat
     2041 gcgtacacgg agcattgtgg ttggatggag ggcggtctgc tgagcgcccg tgaggcaagc
     2101 cgtctgctgt tgcagcgtat cgccgcgtga taggtcatta nggnggnaaa aaaaatgagc
     2161 attctggatt tcccgcgtat ccacttccgt ggctgggccc gtgtcaatgc gccgaccgcg
     2221 aaccgcgatc cgcacggcca catcgatatg gccagcaata ccgtggcgat ggcgggtgag
     2281 ccgttcgacc tggcacgcca tcctacggag ttccaccgtc acctgcgctc cctgggtccg
     2341 cgcttcggct tggatggtcg tgctgacccg gaaggcccgt tcagcctggc cgagggctac
     2401 aacgctgccg gtaacaacca cttttcgtgg gagagcgcaa ccgttagcca cgtgcaatgg
     2461 gatggcggtg aggcggatcg tggtgacggt ctggtcggtg ctcgtttggc actgtggggt
     2521 cactacaatg attatctgcg taccaccttc aatcgtgctc gttgggtcga cagcgacccg
     2581 acgcgccgtg acgctgcaca aatctatgcg ggccaattca ccattagccc ggctggtgcc
     2641 ggtccgggta cgccgtggct gtttacggca gacattgatg atagccatgg tgcacgttgg
     2701 acgcgtggcg gccacattgc agagcgtggc ggccacttct tggatgaaga gtttggtctg
     2761 gcacgcctgt ttcagttctc tgtgccgaaa gatcacccac attttctgtt tcacccgggt
     2821 ccgtttgatt ccgaggcctg gcgtcgtctg caattggctc tggaggatga cgacgttctg
     2881 ggtctgaccg tgcaatatgc gttgttcaat atgagcaccc cgcctcagcc gaacagcccg
     2941 gtttttcacg atatggtcgg tgttgtcggt ctgtggcgtc gtggtgaact ggcgagctac
     3001 ccggctggtc gtctgctgcg tccgcgtcaa ccgggtctgg gtgacctgac cctgcgcgtc
     3061 aacggtggtc gcgttgcgct gaatttggcg tgtgccattc cgttcagcac tcgtgccgcg
     3121 cagccaagcg caccggaccg cctgaccccg gacctgggtg ccaaactgcc gctgggcgat
     3181 ctgctgctgc gtgatgagga cggcgcactg ttggcacgtg tgccgcaggc tctgtaccaa
     3241 gactattgga cgaatcacgg tattgtggac ctgccgctgc tgcgcgaacc gcgtggtagc
     3301 ttgaccctga gcagcgaact ggcggagtgg cgtgagcaag actgggtcac ccaaagcgac
     3361 gcgtctaacc tgtacctgga ggcaccggat cgccgtcacg gtcgcttttt ccctgagagc
     3421 atcgcgctgc gcagctactt tcgcggtgaa gcgcgtgcgc gtccggatat cccgcatcgt
     3481 atcgagggca tgggcctggt cggcgtcgaa tctcgtcagg atggcgacgc tgcggaatgg
     3541 cgtctgacgg gtctgcgtcc gggtccggca cgcattgttc tggacgatgg tgccgaggcg
     3601 atccctctgc gtgttctgcc tgacgattgg gcgctggatg acgcgaccgt cgaagaagtg
     3661 gattacgcct ttttgtaccg ccacgttatg gcgtattacg agctggtgta tccattcatg
     3721 agcgacaagg tgttttccct ggctgatcgt tgcaaatgtg aaacgtacgc acgtctgatg
     3781 tggcagatgt gtgatccgca gaaccgcaac aagtcctatt acatgccgag cacccgcgaa
     3841 ctgtcggcac cgaaagctcg tttgttcttg aagtatctgg cccacgtgga aggccaggca
     3901 cgcctgcaag cacctccgcc agcgggtccg gcacgcattg aatctaaagc ccagttggcg
     3961 gcagagctgc gtaaagccgt cgacctggag ctgtctgtga tgctgcaata cctgtacgcg
     4021 gcgtatagca ttccgaacta tgcacagggc caacaacgtg ttcgtgacgg tgcgtggacc
     4081 gccgagcagc tgcaactggc gtgcggtagc ggtgaccgtc gccgtgatgg cggtattcgt
     4141 gcagcactgc tggaaattgc tcatgaagaa atgattcatt acctggtcgt taacaacctg
     4201 ctgatggccc tgggcgagcc gttctacgcg ggtgtcccgc tgatgggcga agcggcacgt
     4261 caggcgtttg gcctggacac cgagttcgct ctggaaccgt ttagcgaaag cacgctggca
     4321 cgttttgttc gtctggaatg gccgcacttt atcccagcac cgggcaaatc catcgcggac
     4381 tgctatgccg ccattcgtca ggcgtttttg gatctgccgg acttgtttgg tggcgaggca
     4441 ggtaagcgtg gcggtgaaca ccacctgttc ctgaatgagc tgaccaaccg tgcgcatccg
     4501 ggttatcaac tggaagtttt cgatcgcgac tcggcgctgt ttggtattgc atttgtgacc
     4561 gatcagggcg aaggtggcgc tctggacagc ccgcactacg aacatagcca ttttcaacgt
     4621 ctgcgtgaaa tgagcgcgcg tatcatggct caaagcgcac cgttcgaacc ggcgctgccg
     4681 gcgttgcgta atccggttct ggatgagagc ccgggttgcc aacgtgtcgc agacggtcgt
     4741 gcgcgtgcgc tgatggcatt gtaccaaggc gtttatgagc tgatgtttgc gatgatggcg
     4801 cagcacttcg ccgtgaaacc gctgggtagc ttgcgtcgca gccgcctgat gaacgcagca
     4861 atcgatctga tgaccggtct gttgcgtccg ctgagctgcg cgctgatgaa cctgccaagc
     4921 ggcatcgccg gtcgcacggc cggtccgccg ctgccgggtc cggttgacac ccgtagctat
     4981 gacgactacg cgctgggctg tcgcatgctg gcacgccgtt gcgagcgtct gctggagcag
     5041 gcgagcatgc tggaaccggg ttggctgccg gatgcgcaga tggagctgct ggatttctat
     5101 cgtcgccaaa tgctggactt ggcgtgcggc aaactgagcc gcgaggccta ataaggtcca
     5161 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     5221 tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg
     5281 tttatacgct atgtcttcta ctagtagcgg ccgctgcagt ccggcaaaaa agggcaaggt
     5341 gtcaccaccc tgcccttttt ctttaaaacc gaaaagatta cttcgcgtta tgcaggcttc
     5401 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5461 aaaggcggta attcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5521 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5581 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5641 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     5701 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5761 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5821 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5881 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5941 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6001 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6061 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6121 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6181 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtcgat
     6241 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6301 aaagtatata tgagtaaact tggtctgaca gttatcagaa gaactcgtca agaaggcgat
     6361 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
     6421 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
     6481 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
     6541 tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc
     6601 gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
     6661 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
     6721 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
     6781 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
     6841 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
     6901 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca
     6961 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
     7021 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
     7081 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc
     7141 ctcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7201 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7261 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     7321 agaatttcag ataaaaaaaa tccttagctt tcgctaagga tgatttctgg aattcgcggc
     7381 cgcttctaga g
//
