LOCUS       40924_4199        4304 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAJM.336, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4304)
  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 4304)
  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 4304)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4304)
  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..4304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..53
                     /label=L3S3P21
                     /note="L3S3P21"
                     /regulatory_class="terminator"
     promoter        75..103
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    complement(109..128)
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     misc_RNA        130..180
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      212..223
                     /label=B0064
                     /note="B0064"
                     /regulatory_class="ribosome_binding_site"
     CDS             230..946
                     /label=EYFP
                     /note="enhanced YFP"
     regulatory      950..1010
                     /label=L3S2P21
                     /note="L3S2P21"
                     /regulatory_class="terminator"
     regulatory      1015..1068
                     /label=PLacI
                     /note="PLacI"
                     /regulatory_class="promoter"
     regulatory      1033..1038
                     /regulatory_class="minus_35_signal"
     regulatory      1056..1061
                     /regulatory_class="minus_10_signal"
     regulatory      1069..1096
                     /label=lac1
                     /note="lac1"
                     /regulatory_class="ribosome_binding_site"
     CDS             1097..2176
                     /label=lacI
                     /note="lac repressor"
     regulatory      2189..2443
                     /label=induction operon terminator region
                     /note="induction operon terminator region"
                     /regulatory_class="terminator"
     rep_origin      complement(2755..3300)
                     /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(3392..4204)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 ccaattattg aaggcctccc taacgggggg cctttttttg tttctggtct cccgcttaac
       61 gatcgttggc tgtgttgaca attaatcatc ggctcgtata atgtgtggaa ttgtgagcgc
      121 tcacaattag ctgtcaccgg atgtgctttc cggtctgatg agtccgtgag gacgaaacag
      181 cctctacaaa taattttgtt taatactaga gaaagagggg aaatactaga tggtgagcaa
      241 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
      301 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
      361 cctgaagttc atctgcacca caggcaagct gcccgtgccc tggcccaccc tcgtgaccac
      421 cttcggctac ggcctgcaat gcttcgcccg ctaccccgac cacatgaagc tgcacgactt
      481 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
      541 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
      601 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
      661 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
      721 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
      781 gcagaacacc ccaatcggcg acggccccgt gctgctgccc gacaaccact accttagcta
      841 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
      901 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaac tcggtaccaa
      961 attccagaaa agaggcctcc cgaaaggggg gccttttttc gttttggtcc aatggcggcg
     1021 cgccatcgaa tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
     1081 attcagggtg gtgaatatga aaccagtaac gttatacgat gtcgcagagt atgccggtgt
     1141 ctcttatatg accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg
     1201 ggaaaaagtg gaagcggcga tggtggagct gaattacatt cccaaccgcg tggcacaaca
     1261 actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc
     1321 gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt
     1381 ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct
     1441 cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc
     1501 tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc
     1561 catcaacagt attatttact cccatgagga cggtacgcga ctgggcgtgg agcatctggt
     1621 cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg
     1681 tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga
     1741 acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga
     1801 gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg
     1861 cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga
     1921 taccgaagat agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct
     1981 gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg
     2041 caatcagctg ttgccagtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca
     2101 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     2161 actggaaagc gggcagtgat aaggatccta attggtaacg aatcagacaa ttgacggctc
     2221 gagggagtag catagggttt gcagaatccc tgcttcgtcc atttgacagg cacattatgc
     2281 atcgatgata agctgtcaaa catgagcaga tcctctacgc cggacgcatc gtggccggca
     2341 tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag
     2401 atcgggctcg ccacttcggg ctcatgagca aatattttat ctgaggtgct tcctcgctca
     2461 ctgactcgct gcacgaggca gacctcagcg ctagcggagt gtatactggc ttactatgtt
     2521 ggcactgatg agggtgtcag tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg
     2581 tgcgtcagca gaatatgtga tacaggatat attccgcttc ctcgctcact gactcgctac
     2641 gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga acggggcgga gatttcctgg
     2701 aagatgccag gaagatactt aacagggaag tgagagggcc gcggcaaagc cgtttttcca
     2761 taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa
     2821 cccgacagga ctataaagat accaggcgtt tccccctggc ggctccctcg tgcgctctcc
     2881 tgttcctgcc tttcggttta ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc
     2941 cacgcctgac actcagttcc gggtaggcag ttcgctccaa gctggactgt atgcacgaac
     3001 cccccgttca gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3061 aaagacatgc aaaagcacca ctggcagcag ccactggtaa ttgatttaga ggagttagtc
     3121 ttgaagtcat gcgccggtta aggctaaact gaaaggacaa gttttggtga ctgcgctcct
     3181 ccaagccagt tacctcggtt caaagagttg gtagctcaga gaaccttcga aaaaccgccc
     3241 tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa acgatctcaa
     3301 gaagatcatc ttattaaggg gtctgacgct cagtggaacg aaaaatcaat ctaaagtata
     3361 tatgagtaaa cttggtctga cagttacctt agaaaaactc atcgagcatc aaatgaaact
     3421 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     3481 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     3541 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     3601 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca
     3661 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     3721 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt
     3781 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     3841 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg
     3901 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     3961 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     4021 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     4081 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     4141 cagcatccat gttggaattt aatcgcggcc tcgagcaaga cgtttcccgt tgaatatggc
     4201 tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata
     4261 tatttttatc ttgtgcaatg tacatcagag attttgagac acaa
//
