LOCUS       40924_4249        3834 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAJM.847, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3834)
  AUTHORS   Meyer AJ, Segall-Shapiro TH, Voigt CA.
  TITLE     Marionette: E. coli containing 12 highly-optimized small molecule 
            sensors
  JOURNAL   bioRxivorg 285866, 1-26 (2018)
REFERENCE   2  (bases 1 to 3834)
  AUTHORS   Meyer AJ, Segall-Shapiro TH, Voigt CA.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-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 3834)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3834)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxivorg 
            285866, 1-26 (2018)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (21-MAR-2018) Synthetic Biology Center, Department of Biological 
            Engineering, Massachusetts Institute of Technology, 77 Massachusetts
            Avenue, Cambridge, MA 02139, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..53
                     /label=L3S3P21
                     /note="L3S3P21"
                     /regulatory_class="terminator"
     regulatory      73..138
                     /label=PPhlF
                     /note="PPhlF"
                     /regulatory_class="promoter"
     regulatory      104..109
                     /regulatory_class="minus_35_signal"
     misc_feature    109..138
                     /label=PhlO operator
                     /note="PhlO operator"
     regulatory      127..132
                     /regulatory_class="minus_10_signal"
     misc_RNA        140..190
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      222..233
                     /label=B0064
                     /note="B0064"
                     /regulatory_class="ribosome_binding_site"
     CDS             240..956
                     /label=EYFP
                     /note="enhanced YFP"
     regulatory      960..1020
                     /label=L3S2P21
                     /note="L3S2P21"
                     /regulatory_class="terminator"
     regulatory      1025..1078
                     /label=PLacIQ
                     /note="PLacIQ"
                     /regulatory_class="promoter"
     regulatory      1043..1048
                     /regulatory_class="minus_35_signal"
     regulatory      1066..1071
                     /regulatory_class="minus_10_signal"
     regulatory      1079..1106
                     /label=phl2
                     /note="phl2"
                     /regulatory_class="ribosome_binding_site"
     CDS             1107..1709
                     /codon_start=1
                     /product="PhlF-AM"
                     /label=PhlF-AM
                     /protein_id="AYJ72227.1"
                     /translation="MARTPSRSSIGSLRSPHTHKAILTSTIEILKECGYSGLSIESVAR
                     RAGAGKPTIYRWWTNKAALIAEVYENEIEQVRKFPDLGSFKADLDFLLHNLWKVWRETI
                     CGEAFRCVIAEAQLDPVTLTQLKDQFMERRREIPKKLVEDAISNGELPKDINRELLLDM
                     IFGFCWYRLLTEQLTVEQDIEEFTFLLINGVCPGTQC"
     regulatory      1719..1973
                     /label=induction operon terminator region
                     /note="induction operon terminator region"
                     /regulatory_class="terminator"
     rep_origin      complement(2285..2830)
                     /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(2922..3734)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 ccaattattg aaggcctccc taacgggggg cctttttttg tttctggtct cccgcttaac
       61 gatcgttggc tgcgacgtac ggtggaatct gattcgttac caattgacat gatacgaaac
      121 gtaccgtatc gttaaggtag ctgtcaccgg atgtgctttc cggtctgatg agtccgtgag
      181 gacgaaacag cctctacaaa taattttgtt taatactaga gaaagagggg aaatactaga
      241 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
      301 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
      361 gcaagctgac cctgaagttc atctgcacca caggcaagct gcccgtgccc tggcccaccc
      421 tcgtgaccac cttcggctac ggcctgcaat gcttcgcccg ctaccccgac cacatgaagc
      481 tgcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
      541 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
      601 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
      661 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
      721 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
      781 accactacca gcagaacacc ccaatcggcg acggccccgt gctgctgccc gacaaccact
      841 accttagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
      901 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaac
      961 tcggtaccaa attccagaaa agaggcctcc cgaaaggggg gccttttttc gttttggtcc
     1021 aatggcggcg cgccatcgaa tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg
     1081 aagagagtca attcatgggg gtgaatatgg cacgtacccc gagccgtagc agcattggta
     1141 gcctgcgtag tccgcatacc cataaagcaa ttctgaccag caccattgaa atcctgaaag
     1201 aatgtggtta tagcggtctg agcattgaaa gcgtggcacg tcgcgccggt gcaggcaaac
     1261 cgaccattta tcgttggtgg accaacaaag cagcactgat tgccgaagtg tatgaaaatg
     1321 aaatcgaaca ggtacgtaaa tttccggatt tgggtagctt taaagccgat ctggattttc
     1381 tgctgcataa tctgtggaaa gtttggcgtg aaaccatttg tggtgaagca tttcgttgtg
     1441 ttattgcaga agcacagttg gaccctgtaa ccctgaccca actgaaagat cagtttatgg
     1501 aacgtcgtcg tgagataccg aaaaaactgg ttgaagatgc cattagcaat ggtgaactgc
     1561 cgaaagatat caatcgtgaa ctgctgctgg atatgatttt tggtttttgt tggtatcgcc
     1621 tgctgaccga acagttgacc gttgaacagg atattgaaga atttaccttc ctgctgatta
     1681 atggtgtttg tccgggtaca cagtgttgat aaggatccta attggtaacg aatcagacaa
     1741 ttgacggctc gagggagtag catagggttt gcagaatccc tgcttcgtcc atttgacagg
     1801 cacattatgc atcgatgata agctgtcaaa catgagcaga tcctctacgc cggacgcatc
     1861 gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc
     1921 gatggggaag atcgggctcg ccacttcggg ctcatgagca aatattttat ctgaggtgct
     1981 tcctcgctca ctgactcgct gcacgaggca gacctcagcg ctagcggagt gtatactggc
     2041 ttactatgtt ggcactgatg agggtgtcag tgaagtgctt catgtggcag gagaaaaaag
     2101 gctgcaccgg tgcgtcagca gaatatgtga tacaggatat attccgcttc ctcgctcact
     2161 gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga acggggcgga
     2221 gatttcctgg aagatgccag gaagatactt aacagggaag tgagagggcc gcggcaaagc
     2281 cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt
     2341 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctggc ggctccctcg
     2401 tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat tccgctgtta tggccgcgtt
     2461 tgtctcattc cacgcctgac actcagttcc gggtaggcag ttcgctccaa gctggactgt
     2521 atgcacgaac cccccgttca gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2581 tccaacccgg aaagacatgc aaaagcacca ctggcagcag ccactggtaa ttgatttaga
     2641 ggagttagtc ttgaagtcat gcgccggtta aggctaaact gaaaggacaa gttttggtga
     2701 ctgcgctcct ccaagccagt tacctcggtt caaagagttg gtagctcaga gaaccttcga
     2761 aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa
     2821 acgatctcaa gaagatcatc ttattaaggg gtctgacgct cagtggaacg aaaaatcaat
     2881 ctaaagtata tatgagtaaa cttggtctga cagttacctt agaaaaactc atcgagcatc
     2941 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     3001 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     3061 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     3121 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     3181 agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     3241 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     3301 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     3361 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     3421 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     3481 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     3541 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     3601 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     3661 ccatataaat cagcatccat gttggaattt aatcgcggcc tcgagcaaga cgtttcccgt
     3721 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt
     3781 catgatgata tatttttatc ttgtgcaatg tacatcagag attttgagac acaa
//
