LOCUS       40924_4189        3863 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAJM.011, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3863)
  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 3863)
  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 3863)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3863)
  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..3863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..53
                     /label=L3S3P21
                     /note="L3S3P21"
                     /regulatory_class="terminator"
     protein_bind    95..113
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      114..119
                     /regulatory_class="minus_35_signal"
     misc_feature    120..138
                     /label=TetO2 operator
                     /note="TetO2 operator"
     protein_bind    120..138
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      137..142
                     /regulatory_class="minus_10_signal"
     misc_RNA        150..200
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      232..243
                     /label=B0064
                     /note="B0064"
                     /regulatory_class="ribosome_binding_site"
     CDS             250..966
                     /label=EYFP
                     /note="enhanced YFP"
     regulatory      970..1030
                     /label=L3S2P21
                     /note="L3S2P21"
                     /regulatory_class="terminator"
     regulatory      1035..1088
                     /label=PLacI
                     /note="PLacI"
                     /regulatory_class="promoter"
     regulatory      1053..1058
                     /regulatory_class="minus_35_signal"
     regulatory      1076..1081
                     /regulatory_class="minus_10_signal"
     regulatory      1089..1116
                     /label=tet1
                     /note="tet1"
                     /regulatory_class="ribosome_binding_site"
     CDS             1117..1737
                     /label=TetR
                     /note="tetracycline repressor TetR"
     regulatory      1750..2002
                     /label=induction operon terminator region
                     /note="induction operon terminator region"
                     /regulatory_class="terminator"
     rep_origin      complement(2314..2859)
                     /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(2951..3763)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 ccaattattg aaggcctccc taacgggggg cctttttttg tttctggtct cccgcttaac
       61 gatcgttggc tgttttcagc aggacgcact gacctcccta tcagtgatag agattgacat
      121 ccctatcagt gatagagata ctgagcacag ctgtcaccgg atgtgctttc cggtctgatg
      181 agtccgtgag gacgaaacag cctctacaaa taattttgtt taatactaga gaaagagggg
      241 aaatactaga tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
      301 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
      361 gccacctacg gcaagctgac cctgaagttc atctgcacca caggcaagct gcccgtgccc
      421 tggcccaccc tcgtgaccac cttcggctac ggcctgcaat gcttcgcccg ctaccccgac
      481 cacatgaagc tgcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
      541 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
      601 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
      661 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
      721 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
      781 cagctcgccg accactacca gcagaacacc ccaatcggcg acggccccgt gctgctgccc
      841 gacaaccact accttagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
      901 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
      961 tacaagtaac tcggtaccaa attccagaaa agaggcctcc cgaaaggggg gccttttttc
     1021 gttttggtcc aatggcggcg cgccatcgaa tggcgcaaaa cctttcgcgg tatggcatga
     1081 tagcgcccgg aagagagtca attcagggtg gtgaatatgt ccagattaga taaaagtaaa
     1141 gtgattaaca gcgcattaga gctgcttaat gaggtcggaa tcgaaggttt aacaacccgt
     1201 aaactcgccc agaagctagg tgtagagcag cctacattgt attggcatgt aaaaaataag
     1261 cgggctttgc tcgacgcctt agccattgag atgttagata ggcaccatac tcacttttgc
     1321 cctttagaag gggaaagctg gcaagatttt ttacgtaata acgctaaaag ttttagatgt
     1381 gctttactaa gtcatcgcga tggagcaaaa gtacatttag gtacacggcc tacagaaaaa
     1441 cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc aacaaggttt ttcactagag
     1501 aatgcattat atgcactcag cgctgtgggg cattttactt taggttgcgt attggaagat
     1561 caagagcatc aagtcgctaa agaagaaagg gaaacaccta ctactgatag tatgccgcca
     1621 ttattacgac aagctatcga attatttgat caccaaggtg cagagccagc cttcttattc
     1681 ggccttgaat tgatcatatg cggattagaa aaacaactta aatgtgaaag tgggtcctga
     1741 taaggatcct aattggtaac gaatcagaat tgacggctcg agggagtagc atagggtttg
     1801 cagaatccct gcttcgtcca tttgacaggc acattatgca tcgatgataa gctgtcaaac
     1861 atgagcagat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
     1921 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
     1981 tcatgagcaa atattttatc tgaggtgctt cctcgctcac tgactcgctg cacgaggcag
     2041 acctcagcgc tagcggagtg tatactggct tactatgttg gcactgatga gggtgtcagt
     2101 gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat
     2161 acaggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc
     2221 gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta
     2281 acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa
     2341 gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata
     2401 ccaggcgttt ccccctggcg gctccctcgt gcgctctcct gttcctgcct ttcggtttac
     2461 cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc acgcctgaca ctcagttccg
     2521 ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag tccgaccgct
     2581 gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca aaagcaccac
     2641 tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg cgccggttaa
     2701 ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccagtt acctcggttc
     2761 aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt
     2821 tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct tattaagggg
     2881 tctgacgctc agtggaacga aaaatcaatc taaagtatat atgagtaaac ttggtctgac
     2941 agttacctta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
     3001 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     3061 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     3121 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     3181 tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag acttgttcaa
     3241 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     3301 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     3361 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     3421 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
     3481 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     3541 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     3601 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     3661 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     3721 atcgcggcct cgagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac
     3781 tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt
     3841 acatcagaga ttttgagaca caa
//
