LOCUS       V007653                 9286 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V007653
VERSION     V007653
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9286)
  AUTHORS   Cameron DE, Collins JJ.
  TITLE     Tunable protein degradation in bacteria
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9286)
  AUTHORS   Cameron DE, Collins JJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-SEP-2014) Biomedical Engineering, Boston University,
            44 Cummington St., Boston, MA 02215, USA
REFERENCE   3  (bases 1 to 9286)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9286)
  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            "Unpublished"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
            (12-SEP-2014) Biomedical Engineering, Boston University, 44
            Cummington St., Boston, MA 02215, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9286
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..276
                     /label="P32 promoter"
                     /note="P32 promoter"
                     /regulatory_class="promoter"
     regulatory      226..231
                     /label="-35 binding site"
                     /note="-35 binding site"
                     /regulatory_class="minus_35_signal"
     regulatory      249..254
                     /label="-10 binding site"
                     /note="-10 binding site"
                     /regulatory_class="minus_10_signal"
     CDS             304..1011
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     misc_feature    1012..1095
                     /label="pdt#3"
                     /note="pdt#3"
     terminator      1119..1205
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     rep_origin      complement(1370..1958)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     terminator      complement(2046..2140)
                     /label="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     CDS             complement(2173..3030)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(3031..3135)
                     /label="AmpR promoter"
     rep_origin      complement(3167..5714)
                     /direction=LEFT
                     /label="AMB1"
                     /note="AMB1"
     CDS             complement(5735..6469)
                     /gene="ermBP"
                     /label="rRNA adenine N-6-methyltransferase"
                     /note="rRNA adenine N-6-methyltransferase from Enterococcus
                     faecalis. Accession#: P0A4D5"
     terminator      complement(6655..6682)
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             complement(6723..9086)
                     /codon_start=1
                     /product="mf-lon"
                     /label="mf-lon"
                     /protein_id="AIU94531.1"
                     /translation="MSKKIKLPIFQIRGSFIVPGIKENLEVGRKNTLASVNYAIKNSNN
                     QMIAIPQIDASVEKPEFSDLHEFGILIDFEVIKEWKDNSLTISTNPIQRCKVISFFENE
                     DQVPYAEVELIESINDFSDEELKELIEKISDAIKTKASLVTKQIKQLISGESDDLSLAF
                     DSIMFKLAPSKILTNPEYITSPSLKTRWSIIEKIIFAEDGIITRNAESIDAARQKNEIE
                     QELNHKLKEKMDKQQKEYYLREKMRIIKDELEDEDDSDDSSLEKYKERLAKEPFPEEVK
                     RKIMASIKRVEALQSGTPEWNTEKNYIDWMMSIPWWEETEDLTDLKYAKKILDKHHYGM
                     KKVKERIIEYLAVKTKTKSLKAPIITLVGPPGVGKTSLAKSIAEAVGKNFVKVSLGGVK
                     DESEIRGHRKTYVGSMPGRIIQTMKRAKVKNPLFLLDEIDKMASDHRGDPASAMLEVLD
                     PEQNKEFSDHYIEEPYDLSQVMFIATANYPEDIPEALYDRMEIINLSSYTEIEKVKIAQ
                     DYLVPKAIEQHELTSEEISFTEGAINEIIKYYTREAGVRQLERHINSIIRKYIVKNLNG
                     EMDKIVIDEKQVNDLLGKRIFDHTEKQEESQIGVVTGLAYTQFGGDILPIEVSLYPGKG
                     NLILTGKLGEVMKESATIALTYVKSNFEKFGVDKKVFEENDIHVHVPEGAVPKDGPSAG
                     ITITTALISALSDKPVSKEIGMTGEITLRGNVLPIGGLREKSISASRSGLKTIIIPKKN
                     ERDLDEIPDEVKAKLKIIPAEKYEEVFAIVFKTK"
     regulatory      complement(9105..9280)
                     /label="nisA promoter"
                     /note="nisA promoter"
                     /regulatory_class="promoter"
     regulatory      complement(9132..9137)
                     /label="-10 region"
                     /note="-10 region"
                     /regulatory_class="minus_10_signal"
     regulatory      complement(9158..9163)
                     /label="-35 region"
                     /note="-35 region"
                     /regulatory_class="minus_35_signal"
ORIGIN
        1 cgttcgcttg ataaatctcc ctacgggagt ttttgtataa tacccggggt ggcggaactg
       61 gcagacgcac aggacttaaa atcctgcgag gggtaacctt cgtaccggtt cgaaaccggt
      121 cctcgggata tgataagatt aatagtttta gctattaatc tttttttatt tttatttaag
      181 aatggcttaa taaagcggtt actttggatt tttgtgagct tggactagaa aaaaacttca
      241 caaaatgcta tactaggtag gtaaaaaaat attcgggaat tcattaaaga gcgaggtggt
      301 accatggttt ctaaaggtga agaagataat atggctatca tcaaagaatt tatgcgtttc
      361 aaagttcaca tggaaggttc tgttaatggt cacgaatttg aaatcgaagg tgaaggtgaa
      421 ggtcgtcctt acgaaggtac tcaaactgct aaacttaaag ttactaaagg tggtcctctt
      481 cctttcgctt gggatatcct ttctcctcaa ttcatgtacg gttctaaagc ctacgttaaa
      541 caccctgctg atatccctga ttaccttaaa ctttctttcc ctgaaggttt caaatgggaa
      601 cgtgttatga atttcgaaga tggtggtgtt gttactgtta ctcaagattc ttctcttcaa
      661 gatggtgaat ttatctacaa agttaaactt cgtggtacta atttcccttc tgatggtcct
      721 gttatgcaaa aaaaaactat gggttgggaa gcatcttctg aacgtatgta ccctgaagat
      781 ggtgctctta aaggtgaaat caaacaacgt cttaaactta aagatggtgg tcactacgat
      841 gctgaagtta aaactactta caaagctaaa aaacctgttc aacttcctgg tgcttacaat
      901 gttaatatca aacttgatat cacttctcac aatgaagatt acactatcgt tgaacaatac
      961 gaacgtgctg aaggtcgtca ctctactggt ggtatggatg aactttacaa agcggcgaac
     1021 aaaaacgaag aaaacaccaa cgaagtgccg acctttatgc tgaacgcggg ccaggcgaac
     1081 agaagacgag tttaatctag aggttccacg cgtggcatca aataaaacga aaggctcagt
     1141 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     1201 caaatccgcc gccctagacc tagggcgttc ggctgcggcg agcggtatca gctcactcaa
     1261 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1321 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1381 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1441 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1501 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1561 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1621 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1681 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1741 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1801 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1861 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1921 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1981 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgactagtg
     2041 cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga
     2101 tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct cgtaaacttg
     2161 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2221 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     2281 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     2341 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     2401 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     2461 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     2521 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     2581 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     2641 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     2701 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     2761 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     2821 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     2881 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     2941 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3001 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3061 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3121 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gcatgcgatt ttcctcctct
     3181 aatatgctca acttaaatga cctattcaat aaatctatta tgctgctaaa tagtttatag
     3241 gacaaataag tatactctaa tgacctataa aagatagaaa attaaaaaat caagtgttcg
     3301 cttcgctctc actgcccctc gacgttttag tagcctttcc ctcacttcgt tcagtccaag
     3361 ccaactaaaa gttttcgggc tactctctcc ttctccccct aataattaat taaaatctta
     3421 ctctgtatat ttctgctaat cattcgctaa acagcaaaga aaaaacaaac acgtatcata
     3481 gatataaatg taatggcata gtgcgggttt tattttcagc ctgtatcata gctaaacaaa
     3541 tcgagttgtg tgtccgtttt agggcgttct gctagcttgt ttaaagtctc ttgaatgaat
     3601 gtatgctcta agtcaaaaga atttgtcagc gcctttatat agctttcttt ttcttctttt
     3661 tttactttaa tgatcgatag caacaatgat ttaacactag caagttgaat gccaccattt
     3721 cttcctggtt taatcttaaa gaaaatttcc tgattcgcct tcagtacctt cagcaattta
     3781 tctaatgtcc gttcaggaat gcctagcact tctctaatct cttttttggt cgtcactaaa
     3841 taaggcttgt atacatcgct tttttcgcta atataagcca ttaaatcttc tttccattct
     3901 gacaaatgaa cacgttgacg ttcgcttctt tttttcttga atttaaacca cccttgacgg
     3961 acaaataaat ctttactggt taaatcactt gatacccaag ctttgcaaag aatggtaatg
     4021 tattccctat tagccccttg atagttttct gaataggcac ttctaacaat tttgattact
     4081 tctttttctt ctaagggttg atctaatcga ttattaaact caaacatatt atattcgcac
     4141 gtttcgattg aatagcctga actaaagtag gctaaagaga gggtaaacat gacgttatta
     4201 cgccctatta aacccttttc tcctgaaaat ttcgtttcgt gcaataagag attaaaccag
     4261 ggttcatcta cttgtttttt gccttctgta ccgcttaaaa ccgttagact tgaacgagta
     4321 aagcccttat tatctgtttg tttgaaagac caatcttgcc attctttgaa agaataacgg
     4381 taattaggat caaaaaattc tacattgtcc gttcttggta tgcgagcaat accaaaatga
     4441 ttacacgtta gatcaactgg caaagacttt ccaaaatatt ctcggatatt ttgcgaaatt
     4501 attttggctg ctttgacaga tttaaattct gattttgaag tcacatagac tggcgtttct
     4561 aaaacaaaat atgcttgata acctttatca gatttgataa tcatagtagg cataaaacct
     4621 aaatcaatag cggttgttaa aatatcgctt gctgaaatag tttcttttgc cgtgtgaata
     4681 tcaaaatcaa taaagaaggt attgatttgt cttaaattgt tttcagaatg tcctttcgtg
     4741 tatgaacggt tttcgtctgc atacgttcca taacgataaa cgttgggtgt ccaatgtgta
     4801 aatgtatctt gattttcttg aatcgcttcc tcggaagtca gaacaacacc acgaccgcca
     4861 atcatgcttg attttgagcg atacgcaaaa atagcccctt tgcttttacc tggcttggta
     4921 gtgattgagc gaattttact atttttaaat ttgtacttta acaagccgtc atgaagcaca
     4981 gtttctacaa caaaagggat attcattcag ctgttctcct ttcctataaa tcctataaaa
     5041 taggttgttt aattaacttg gtttgctttt tcattcaact gtttcaatat tgcatgtttt
     5101 gaaaaagatt tttttccttt ataagtcaat ttttttccac taatcgaata aattattttg
     5161 ttattttcta ttaacttata tatataatct tccccctccg aagaaaaata cttatctgat
     5221 tttgtttcta agtagatatt tctcttttct aactctttct taaacgtttc tagtgtatag
     5281 atatttgcta attttcttat ctccaataaa ctatttttta tataagtttt acattcatca
     5341 tgattcatac aaactccacc ttctataaat gaatacaaaa aaagcaatca aacgatttcc
     5401 gattgattgc ttaacaattc ttaaattcag tagcttagat acttgaaaac tctctgattt
     5461 ccctatataa tgatagtacg gttatatacc gtcttcaaac aaagttaatt aaataacttc
     5521 ttacgaggga agagttcatc tgactaactg ataagcgttg gtttggcaat cttatcgggc
     5581 tatgcattta taaaatgtcg tcaaacattt tataaatgtg tcatggctct tttttcgttt
     5641 ctattcagtt cgttgtttcg ttatatctag tataccgctt ttaaaaaaaa ataagcaacg
     5701 atttcgtgca ttataccggt gactcataga attatttcct cccgttaaat aatagataac
     5761 tattaaaaat agacaatact tgctcataag taacggtact taaattgttt actttggcgt
     5821 gtttcattgc ttgatgaaac tgatttttag taaacagttg acgatattct cgattgaccc
     5881 attttgaaac aaagtacgta tatagcttcc aatatttatc tggaacatct gtggtatggc
     5941 gggtaagttt tattaagaca ctgtttactt ttggtttagg atgaaagcat tccgctggca
     6001 gcttaagcaa ttgctgaatc gagacttgag tgtgcaagag caaccctagt gttcggtgaa
     6061 tatccaaggt acgcttgtag aatccttctt caacaatcag atagatgtca gacgcatggc
     6121 tttcaaaaac cactttttta ataatttgtg tgcttaaatg gtaaggaata ctcccaacaa
     6181 ttttatacct ctgtttgtta gggaattgaa actgtagaat atcttggtga attaaagtga
     6241 cacgagtatt cagttttaat ttttctgacg ataagttgaa tagatgactg tctaattcaa
     6301 tagacgttac ctgtttactt attttagcca gtttcgtcgt taaatgccct ttacctgttc
     6361 caatttcgta aacggtatcg gtttctttta aattcaattg ttttattatt tggttgagta
     6421 ctttttcact cgttaaaaag ttttgagaat attttatatt tttgttcatg taatcactcc
     6481 ttcttaatta caaattttta gcatctaatt taacttcaat tcctattata caaaatttta
     6541 agatactgca ctatcaacac actcttaagt ttgcttctaa gtcttatttc cataacttct
     6601 tttacgtttc cgcaggctgc aggcggatcc ccaacaagag tttgtagaaa cgcaaaaagg
     6661 ccatccgtca ggatggcctt ctgcttagct agagcggcgg atttgtccta ctcagggacg
     6721 tcttattttg ttttaaaaac tattgcaaat acctcttcat atttttcagc aggtataatt
     6781 tttagttttg cttttacttc atctggaatt tcgtctaagt ctctttcatt tttctttgga
     6841 ataatgattg ttttcaatcc acttcttgaa gcagaaattg atttttctct taatccacca
     6901 attggcaata catttcctct tagtgtgatt tcaccagtca ttccaatttc ttttgaaact
     6961 ggtttatctg aaagtgcaga aattaaagca gttgtaattg tgataccagc acttggtcca
     7021 tcttttggaa cagctccttc aggaacgtga acatgaatat cattttcttc aaatactttt
     7081 ttatctactc caaatttttc aaagtttgat ttaacataag ttaacgcaat tgttgcagat
     7141 tctttcataa cttcaccaag tttacctgta agaattaaat ttcctttacc agggtataaa
     7201 cttacttcaa ttggtaaaat atctccacca aattgtgtat atgctaaacc tgtaaccaca
     7261 ccaatttgag attcttcttg tttttcagtg tgatcaaaaa ttcttttacc taataaatca
     7321 tttacttgtt tttcatcaat aacaatttta tccatttcac catttagatt tttaacaata
     7381 tactttctaa taatagaatt aatatgtctt tctaattgac gcacaccagc ttctctagtg
     7441 taatatttaa taatttcatt aattgcacct tcagtaaatg atatttcttc tgaagttaat
     7501 tcatgttgtt ctattgcttt tggaactaaa taatcttgtg ctatcttaac tttttcaatt
     7561 tcagtatagc tagataaatt tatgatttcc atacggtcat ataaggcttc aggaatatct
     7621 tcaggatagt tagcagttgc tataaacata acttggctta gatcatatgg ttcttcaata
     7681 taatgatctg aaaactcttt gttttgttct gggtcaagaa cctctaacat tgcacttgct
     7741 ggatcacctc tgtgatcaga tgccatttta tcaatttcat caagcaagaa taatgggttt
     7801 tttacttttg ctcttttcat tgtttgaata attcttcctg gcattgaacc aacataagtt
     7861 tttctatgtc cacgaatttc tgattcatct ttaactccac ctaaacttac tttaacaaag
     7921 ttttttccaa ctgcttctgc aattgatttt gctaaacttg ttttacctac acctggggga
     7981 ccaactaatg taataattgg tgcttttagt gattttgttt tagttttaac agctaagtac
     8041 tcaataattc tttctttaac ttttttcata ccataatgat gtttgtctaa aattttttta
     8101 gcatatttta aatcagttaa gtcttcagtt tcttcccacc agggaatact catcatccaa
     8161 tcaatgtaat ttttctcagt attccattca ggagttcctg attgaagtgc ttcaactcgt
     8221 ttgatagaag ccataatttt tcgtttgact tcttcaggga aaggttcttt agctaaacgt
     8281 tctttatatt tttctaatga actgtcatca gaatcatctt catcttctaa ttcatctttg
     8341 ataattcgca ttttttctcg cagataatat tctttttgtt gtttatccat tttttctttt
     8401 aatttatggt tcagttcttg ttcaatttca ttcttttgtc gtgctgcatc aattgattct
     8461 gcatttcttg ttatgattcc atcttcagca aaaattattt tttcaataat tgaccatcgt
     8521 gtttttaatg acggagatgt tatgtattcc ggatttgtca atatttttga cggagccagt
     8581 ttaaacataa ttgaatcaaa agctaaactt aaatcatcag attcacctga tattaactgt
     8641 ttaatttgtt ttgttactaa actagcttta gttttgatag catcagagat tttttcaatt
     8701 aattctttta actcttcatc actaaaatca ttaattgatt caattaattc aacttccgca
     8761 tacggcactt gatcttcatt ttcaaaaaag ctaattactt tacatcgttg aatagggttt
     8821 gtgctaatag ttaatgaatt atctttccat tctttgatta cttcaaaatc aattaaaatt
     8881 ccaaattcat gtaaatctga aaattcaggt ttttcaacag atgcatcaat ctgcggtatt
     8941 gcgatcattt gattatttga atttttaatc gcataattaa cgctcgctaa tgtgtttttt
     9001 cttccaactt ctaaattctc tttaattcca ggaacaataa atgaaccgcg tatttggaaa
     9061 ataggtagtt ttattttttt actcatgcta gcccctaacc gcggtatttt gtagttcctt
     9121 cgaacgaaat cattgtatct aacaaacttc agaatttaat cagagccgtt tattatgctc
     9181 gcgttatcga caataatatt attaccaata ctttctcaag atagaattaa gactgtttta
     9241 gatttgttaa tgtttctatt gtcagtatag ttataagact ctcgag
//
