LOCUS       V006489                 8684 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V006489
VERSION     V006489
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8684)
  AUTHORS   Pardee K, Slomovic S, Nguyen PQ, Lee JW, Donghia N, Burrill D,
            Ferrante T, McSorley F, Furuta Y, Lewandowski M, Boddy CN, Joshi NS,
            Collins JJ.
  TITLE     Portable, On-demand Biomolecular Manufacturing
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8684)
  AUTHORS   Pardee K, Slomovic S, Nguyen PQ, Lee JW, Donghia N, Burrill D,
            Ferrante T, McSorley F, Furuta Y, Lewandowski M, Boddy CN, Joshi NS,
            Collins JJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JUN-2016) Wyss Institute, Harvard University, 3
            Blackfan Circle, Wyss Institute Rm517-1D, Boston, MA 02115, USA
REFERENCE   3  (bases 1 to 8684)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8684)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (29-JUN-2016) Wyss Institute, Harvard University, 3 Blackfan Circle,
            Wyss Institute Rm517-1D, Boston, MA 02115, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        104..208
                     /label="AmpR promoter"
     CDS             209..1066
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1240..1828
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(2014..2156)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(2261..2449)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(3727..3748)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3764..4843)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(4844..4921)
                     /label="lacI promoter"
     promoter        5230..5248
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5249..5273
                     /label="lac operator"
                     /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             5288..5310
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5317..8310
                     /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"
     CDS             8320..8343
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     terminator      8425..8472
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     promoter        complement(8647..8675)
                     /label="tet promoter"
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
ORIGIN
        1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
       61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      181 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
      481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
      541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
      601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
      661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
      721 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
      781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
      841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
      901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
      961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     1921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     1981 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     2041 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc
     2101 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     2161 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2221 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2281 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     2341 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     2401 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     2461 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     2521 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     2581 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     2641 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     2701 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     2761 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     2821 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     2881 tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga
     2941 tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg
     3001 tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc
     3061 cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg
     3121 acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt
     3181 ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccag tgatcgaagt
     3241 taggctggta agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg
     3301 cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat
     3361 aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc
     3421 ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt
     3481 gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat
     3541 cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg
     3601 tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg
     3661 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg
     3721 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3781 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3841 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
     3901 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
     3961 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
     4021 cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc
     4081 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     4141 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     4201 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     4261 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
     4321 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
     4381 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     4441 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     4501 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
     4561 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     4621 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     4681 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     4741 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     4801 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     4861 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt
     4921 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     4981 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     5041 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     5101 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     5161 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
     5221 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
     5281 aaataatttt gtttaacttt aagaaggaga tataccatga gcattctgga tttcccgcgt
     5341 atccacttcc gtggctgggc ccgtgtcaat gcgccgaccg cgaaccgcga tccgcacggc
     5401 cacatcgata tggccagcaa taccgtggcg atggcgggtg agccgttcga cctggcacgc
     5461 catcctacgg agttccaccg tcacctgcgc tccctgggtc cgcgcttcgg cttggatggt
     5521 cgtgctgacc cggaaggccc gttcagcctg gccgagggct acaacgctgc cggtaacaac
     5581 cacttttcgt gggagagcgc aaccgttagc cacgtgcaat gggatggcgg tgaggcggat
     5641 cgtggtgacg gtctggtcgg tgctcgtttg gcactgtggg gtcactacaa tgattatctg
     5701 cgtaccacct tcaatcgtgc tcgttgggtc gacagcgacc cgacgcgccg tgacgctgca
     5761 caaatctatg cgggccaatt caccattagc ccggctggtg ccggtccggg tacgccgtgg
     5821 ctgtttacgg cagacattga tgatagccat ggtgcacgtt ggacgcgtgg cggccacatt
     5881 gcagagcgtg gcggccactt cttggatgaa gagtttggtc tggcacgcct gtttcagttc
     5941 tctgtgccga aagatcaccc acattttctg tttcacccgg gtccgtttga ttccgaggcc
     6001 tggcgtcgtc tgcaattggc tctggaggat gacgacgttc tgggtctgac cgtgcaatat
     6061 gcgttgttca atatgagcac cccgcctcag ccgaacagcc cggtttttca cgatatggtc
     6121 ggtgttgtcg gtctgtggcg tcgtggtgaa ctggcgagct acccggctgg tcgtctgctg
     6181 cgtccgcgtc aaccgggtct gggtgacctg accctgcgcg tcaacggtgg tcgcgttgcg
     6241 ctgaatttgg cgtgtgccat tccgttcagc actcgtgccg cgcagccaag cgcaccggac
     6301 cgcctgaccc cggacctggg tgccaaactg ccgctgggcg atctgctgct gcgtgatgag
     6361 gacggcgcac tgttggcacg tgtgccgcag gctctgtacc aagactattg gacgaatcac
     6421 ggtattgtgg acctgccgct gctgcgcgaa ccgcgtggta gcttgaccct gagcagcgaa
     6481 ctggcggagt ggcgtgagca agactgggtc acccaaagcg acgcgtctaa cctgtacctg
     6541 gaggcaccgg atcgccgtca cggtcgcttt ttccctgaga gcatcgcgct gcgcagctac
     6601 tttcgcggtg aagcgcgtgc gcgtccggat atcccgcatc gtatcgaggg catgggcctg
     6661 gtcggcgtcg aatctcgtca ggatggcgac gctgcggaat ggcgtctgac gggtctgcgt
     6721 ccgggtccgg cacgcattgt tctggacgat ggtgccgagg cgatccctct gcgtgttctg
     6781 cctgacgatt gggcgctgga tgacgcgacc gtcgaagaag tggattacgc ctttttgtac
     6841 cgccacgtta tggcgtatta cgagctggtg tatccattca tgagcgacaa ggtgttttcc
     6901 ctggctgatc gttgcaaatg tgaaacgtac gcacgtctga tgtggcagat gtgtgatccg
     6961 cagaaccgca acaagtccta ttacatgccg agcacccgcg aactgtcggc accgaaagct
     7021 cgtttgttct tgaagtatct ggcccacgtg gaaggccagg cacgcctgca agcacctccg
     7081 ccagcgggtc cggcacgcat tgaatctaaa gcccagttgg cggcagagct gcgtaaagcc
     7141 gtcgacctgg agctgtctgt gatgctgcaa tacctgtacg cggcgtatag cattccgaac
     7201 tatgcacagg gccaacaacg tgttcgtgac ggtgcgtgga ccgccgagca gctgcaactg
     7261 gcgtgcggta gcggtgaccg tcgccgtgat ggcggtattc gtgcagcact gctggaaatt
     7321 gctcatgaag aaatgattca ttacctggtc gttaacaacc tgctgatggc cctgggcgag
     7381 ccgttctacg cgggtgtccc gctgatgggc gaagcggcac gtcaggcgtt tggcctggac
     7441 accgagttcg ctctggaacc gtttagcgaa agcacgctgg cacgttttgt tcgtctggaa
     7501 tggccgcact ttatcccagc accgggcaaa tccatcgcgg actgctatgc cgccattcgt
     7561 caggcgtttt tggatctgcc ggacttgttt ggtggcgagg caggtaagcg tggcggtgaa
     7621 caccacctgt tcctgaatga gctgaccaac cgtgcgcatc cgggttatca actggaagtt
     7681 ttcgatcgcg actcggcgct gtttggtatt gcatttgtga ccgatcaggg cgaaggtggc
     7741 gctctggaca gcccgcacta cgaacatagc cattttcaac gtctgcgtga aatgagcgcg
     7801 cgtatcatgg ctcaaagcgc accgttcgaa ccggcgctgc cggcgttgcg taatccggtt
     7861 ctggatgaga gcccgggttg ccaacgtgtc gcagacggtc gtgcgcgtgc gctgatggca
     7921 ttgtaccaag gcgtttatga gctgatgttt gcgatgatgg cgcagcactt cgccgtgaaa
     7981 ccgctgggta gcttgcgtcg cagccgcctg atgaacgcag caatcgatct gatgaccggt
     8041 ctgttgcgtc cgctgagctg cgcgctgatg aacctgccaa gcggcatcgc cggtcgcacg
     8101 gccggtccgc cgctgccggg tccggttgac acccgtagct atgacgacta cgcgctgggc
     8161 tgtcgcatgc tggcacgccg ttgcgagcgt ctgctggagc aggcgagcat gctggaaccg
     8221 ggttggctgc cggatgcgca gatggagctg ctggatttct atcgtcgcca aatgctggac
     8281 ttggcgtgcg gcaaactgag ccgcgaggcc actagtggtg actacaaaga cgacgacgac
     8341 aaataaccta ggtaactcga ggatccggct gctaacaaag cccgaaagga agctgagttg
     8401 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     8461 aggggttttt tgctgaaagg aggaactata tccggatatc ccgcaagagg cccggcagta
     8521 ccggcataac caagcctatg cctacagcat ccagggtgac ggtgccgagg atgacgatga
     8581 gcgcattgtt agatttcata cacggtgcct gactgcgtta gcaatttaac tgtgataaac
     8641 taccgcatta aagcttatcg atgataagct gtcaaacatg agaa
//
