LOCUS       40924_4194        4143 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAJM.1642, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4143)
  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 4143)
  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 4143)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4143)
  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..4143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..53
                     /label=L3S3P21
                     /note="L3S3P21"
                     /regulatory_class="terminator"
     regulatory      73..298
                     /label=PCin
                     /note="PCin"
                     /regulatory_class="promoter"
     misc_feature    225..245
                     /label=CinO operator
                     /note="CinO operator"
     regulatory      246..251
                     /regulatory_class="minus_35_signal"
     regulatory      269..274
                     /regulatory_class="minus_10_signal"
     misc_RNA        300..350
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      382..393
                     /label=B0064
                     /note="B0064"
                     /regulatory_class="ribosome_binding_site"
     CDS             400..1116
                     /label=EYFP
                     /note="enhanced YFP"
     regulatory      1120..1180
                     /label=L3S2P21
                     /note="L3S2P21"
                     /regulatory_class="terminator"
     regulatory      1185..1238
                     /label=PLacI
                     /note="PLacI"
                     /regulatory_class="promoter"
     regulatory      1203..1208
                     /regulatory_class="minus_35_signal"
     regulatory      1226..1231
                     /regulatory_class="minus_10_signal"
     regulatory      1239..1266
                     /label=cin1
                     /note="cin1"
                     /regulatory_class="ribosome_binding_site"
     CDS             1267..2022
                     /codon_start=1
                     /product="CinR-AM"
                     /label=CinR-AM
                     /protein_id="AYJ72261.1"
                     /translation="MIENTYSEKFESAFEQIKAAANVDAAIRILQAEYNLDFVTYHLAQ
                     TIASKIDSPFVRTTYPDAWVSRYLLNCYVKVDPIIKQGFERQLPFDWSEVEPTPEAYAM
                     LVDAQKHGIDDNGYSIPVADKAQRRALLSLNAHIPADEWTELVRRCRNEWIEIAHLIHR
                     KAVYELHGENDPVPALSPREIECLHWTALGKDYKDISVILGISEHTTRDYLKTARFRLG
                     CTTISAAASRAVQLRIINPYRIRMTRRNW"
     regulatory      2028..2282
                     /label=induction operon terminator region
                     /note="induction operon terminator region"
                     /regulatory_class="terminator"
     rep_origin      complement(2594..3139)
                     /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(3231..4043)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 ccaattattg aaggcctccc taacgggggg cctttttttg tttctggtct cccgcttaac
       61 gatcgttggc tgccctttgt gcgtccaaac ggacgcacgg cgctctaaag cgggtcgcga
      121 tctttcagat tcgctcctcg cgctttcagt ctttgttttg gcgcatgtcg ttatcgcaaa
      181 accgctgcac acttttgcgc gacatgctct gatccccctc atctgggggg gcctatctga
      241 gggaatttcc gatccggctc gcctgaacca ttctgctttc cacgaacttg aaaacgctag
      301 ctgtcaccgg atgtgctttc cggtctgatg agtccgtgag gacgaaacag cctctacaaa
      361 taattttgtt taatactaga gaaagagggg aaatactaga tggtgagcaa gggcgaggag
      421 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      481 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
      541 atctgcacca caggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac
      601 ggcctgcaat gcttcgcccg ctaccccgac cacatgaagc tgcacgactt cttcaagtcc
      661 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
      721 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
      781 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
      841 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
      901 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
      961 ccaatcggcg acggccccgt gctgctgccc gacaaccact accttagcta ccagtccgcc
     1021 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1081 gccgggatca ctctcggcat ggacgagctg tacaagtaac tcggtaccaa attccagaaa
     1141 agaggcctcc cgaaaggggg gccttttttc gttttggtcc aatggcggcg cgccatcgaa
     1201 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
     1261 gtgaatatga ttgagaatac ctatagcgaa aagttcgagt ccgcgttcga acagatcaaa
     1321 gcggcggcca acgtggatgc cgccatccgt attctccagg cggaatataa cctcgatttc
     1381 gtcacctacc atctcgccca gacaatcgcg agcaagatcg attcgccctt cgtgcgcacc
     1441 acctatccgg atgcctgggt ttcccgttac ctcctcaact gctatgtgaa ggtcgatccg
     1501 atcatcaagc agggcttcga acgccagctg cccttcgact ggagcgaggt cgaaccgacg
     1561 ccggaggcct atgccatgct ggtcgacgcc cagaaacacg gcatcgatga caatggctac
     1621 tccatccccg tcgccgacaa ggcgcagcgc cgcgccctgc tgtcgctgaa tgcccatata
     1681 ccggccgacg aatggaccga gctcgtgcgc cgctgccgca atgagtggat cgagatcgcc
     1741 catctgatcc accgcaaggc cgtatatgag ctgcatggcg aaaacgatcc ggtgccggca
     1801 ttgtcgccgc gcgagatcga gtgtctgcac tggaccgccc tcggcaagga ttacaaggat
     1861 atttcggtca tcctgggcat atcagagcat accacacgcg attacctgaa aaccgcccgc
     1921 ttcaggctcg gctgcaccac gatctcggcc gccgcgtcgc gggctgttca attgcgcatc
     1981 atcaatccct ataggatccg catgacgcga cgtaattggt aatagcctaa ttggtaacga
     2041 atcagacaat tgacggctcg agggagtagc atagggtttg cagaatccct gcttcgtcca
     2101 tttgacaggc acattatgca tcgatgataa gctgtcaaac atgagcagat cctctacgcc
     2161 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
     2221 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcaa atattttatc
     2281 tgaggtgctt cctcgctcac tgactcgctg cacgaggcag acctcagcgc tagcggagtg
     2341 tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc atgtggcagg
     2401 agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata ttccgcttcc
     2461 tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa
     2521 cggggcggag atttcctgga agatgccagg aagatactta acagggaagt gagagggccg
     2581 cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct
     2641 caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg
     2701 gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat
     2761 ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag
     2821 ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat
     2881 cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat
     2941 tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag
     3001 ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag
     3061 aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg
     3121 cagaccaaaa cgatctcaag aagatcatct tattaagggg tctgacgctc agtggaacga
     3181 aaaatcaatc taaagtatat atgagtaaac ttggtctgac agttacctta gaaaaactca
     3241 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     3301 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     3361 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     3421 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     3481 aatggcaaaa gcttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     3541 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     3601 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     3661 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     3721 tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     3781 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     3841 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     3901 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     3961 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct cgagcaagac
     4021 gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt
     4081 tttattgttc atgatgatat atttttatct tgtgcaatgt acatcagaga ttttgagaca
     4141 caa
//
