LOCUS       40924_4209        3867 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAJM.657, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3867)
  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 3867)
  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 3867)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3867)
  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..3867
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..53
                     /label=L3S3P21
                     /note="L3S3P21"
                     /regulatory_class="terminator"
     protein_bind    73..100
                     /label=CuO
                     /note="CymR-binding P2 operator sequence from the p-cmt
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     regulatory      119..124
                     /regulatory_class="minus_10_signal"
     misc_feature    131..162
                     /label=CymO operator
                     /note="CymO operator"
     protein_bind    131..158
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     regulatory      131
                     /label=+1
                     /note="+1"
                     /regulatory_class="other"
     misc_RNA        164..214
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      246..257
                     /label=B0064
                     /note="B0064"
                     /regulatory_class="ribosome_binding_site"
     CDS             264..980
                     /label=EYFP
                     /note="enhanced YFP"
     regulatory      984..1044
                     /label=L3S2P21
                     /note="L3S2P21"
                     /regulatory_class="terminator"
     regulatory      1049..1102
                     /label=PLacIQ
                     /note="PLacIQ"
                     /regulatory_class="promoter"
     regulatory      1067..1072
                     /regulatory_class="minus_35_signal"
     regulatory      1090..1095
                     /regulatory_class="minus_10_signal"
     regulatory      1103..1130
                     /label=cym1
                     /note="cym1"
                     /regulatory_class="ribosome_binding_site"
     CDS             1131..1739
                     /label=CymR
                     /note="cumate repressor (Mullick et al., 2006)"
     regulatory      1752..2006
                     /label=induction operon terminator region
                     /note="induction operon terminator region"
                     /regulatory_class="terminator"
     rep_origin      complement(2318..2863)
                     /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(2955..3767)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 ccaattattg aaggcctccc taacgggggg cctttttttg tttctggtct cccgcttaac
       61 gatcgttggc tgaacaaaca gacaatctgg tctgtttgta ttatggaaaa tttttctgta
      121 taatagattc aacaaacaga caatctggtc tgtttgtatt atagctgtca ccggatgtgc
      181 tttccggtct gatgagtccg tgaggacgaa acagcctcta caaataattt tgtttaatac
      241 tagagaaaga ggggaaatac tagatggtga gcaagggcga ggagctgttc accggggtgg
      301 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
      361 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accacaggca
      421 agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg caatgcttcg
      481 cccgctaccc cgaccacatg aagctgcacg acttcttcaa gtccgccatg cccgaaggct
      541 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
      601 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
      661 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
      721 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
      781 aggacggcag cgtgcagctc gccgaccact accagcagaa caccccaatc ggcgacggcc
      841 ccgtgctgct gcccgacaac cactacctta gctaccagtc cgccctgagc aaagacccca
      901 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
      961 gcatggacga gctgtacaag taactcggta ccaaattcca gaaaagaggc ctcccgaaag
     1021 gggggccttt tttcgttttg gtccaatggc ggcgcgccat cgaatggtgc aaaacctttc
     1081 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat atgagcccga
     1141 aacgtcgtac ccaggcagaa cgtgcaatgg aaacccaggg taaactgatt gcagcagcac
     1201 tgggtgttct gcgtgaaaaa ggttatgcag gttttcgtat tgcagatgtt ccgggtgcag
     1261 ccggtgttag ccgtggtgca cagagccatc attttccgac caaactggaa ctgctgctgg
     1321 caacctttga atggctgtat gagcagatta ccgaacgtag ccgtgcacgt ctggcaaaac
     1381 tgaaaccgga agatgatgtt attcagcaga tgctggatga tgcagcagaa ttttttctgg
     1441 atgatgattt tagcatcggc ctggatctga ttgttgcagc agatcgtgat ccggcactgc
     1501 gtgaaggtat tcagcgtacc gttgaacgta atcgttttgt tgttgaagat atgtggctgg
     1561 gtgtgctggt gagccgtggt ctgagccgtg atgatgccga agatattctg tggctgattt
     1621 ttaacagcgt tcgtggtctg gtagttcgta gcctgtggca gaaagataaa gaacgttttg
     1681 aacgtgtgcg taatagcacc ctggaaattg cacgtgaacg ttatgcaaaa ttcaaacgtt
     1741 gataaggatc ctaattggta acgaatcaga caattgacgg ctcgagggag tagcataggg
     1801 tttgcagaat ccctgcttcg tccatttgac aggcacatta tgcatcgatg ataagctgtc
     1861 aaacatgagc agatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt
     1921 gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc
     1981 gggctcatga gcaaatattt tatctgaggt gcttcctcgc tcactgactc gctgcacgag
     2041 gcagacctca gcgctagcgg agtgtatact ggcttactat gttggcactg atgagggtgt
     2101 cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac cggtgcgtca gcagaatatg
     2161 tgatacagga tatattccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg
     2221 cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc caggaagata
     2281 cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc cgcccccctg
     2341 acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca ggactataaa
     2401 gataccaggc gtttccccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt
     2461 ttaccggtgt cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt
     2521 tccgggtagg cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac
     2581 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca
     2641 ccactggcag cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg
     2701 ttaaggctaa actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg
     2761 gttcaaagag ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc
     2821 gttttcagag caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa
     2881 ggggtctgac gctcagtgga acgaaaaatc aatctaaagt atatatgagt aaacttggtc
     2941 tgacagttac cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     3001 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     3061 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     3121 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     3181 tgagtgacga ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt
     3241 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     3301 attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa
     3361 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     3421 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt
     3481 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     3541 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     3601 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
     3661 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     3721 tttaatcgcg gcctcgagca agacgtttcc cgttgaatat ggctcataac accccttgta
     3781 ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca
     3841 atgtacatca gagattttga gacacaa
//
