LOCUS       40924_4229        4098 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAJM.690, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4098)
  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 4098)
  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 4098)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4098)
  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..4098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..53
                     /label=L3S3P21
                     /note="L3S3P21"
                     /regulatory_class="terminator"
     regulatory      73..165
                     /label=P3B5B
                     /note="P3B5B"
                     /regulatory_class="promoter"
     misc_feature    73..118
                     /label=PcaO operator
                     /note="PcaO operator"
     regulatory      129..134
                     /regulatory_class="minus_35_signal"
     regulatory      153..158
                     /regulatory_class="minus_10_signal"
     misc_RNA        167..217
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      249..260
                     /label=B0064
                     /note="B0064"
                     /regulatory_class="ribosome_binding_site"
     CDS             267..983
                     /label=EYFP
                     /note="enhanced YFP"
     regulatory      987..1047
                     /label=L3S2P21
                     /note="L3S2P21"
                     /regulatory_class="terminator"
     regulatory      1052..1105
                     /label=PLacIQ
                     /note="PLacIQ"
                     /regulatory_class="promoter"
     regulatory      1070..1075
                     /regulatory_class="minus_35_signal"
     regulatory      1093..1098
                     /regulatory_class="minus_10_signal"
     regulatory      1106..1133
                     /label=pca2
                     /note="pca2"
                     /regulatory_class="ribosome_binding_site"
     CDS             1134..1970
                     /codon_start=1
                     /product="PcaU-AM"
                     /label=PcaU-AM
                     /protein_id="AYJ72255.1"
                     /translation="MWSNMDDKKVKEENILHNSTNKKIIRHEDFVAGISKGMAILDSFG
                     TDRHRLNITMAAEKTGMTRAAARRHLLTLEYLGYLESDGHYFYLTPKILKFSGSYLGGA
                     QLPKISQPLLNLLTTQTSLIYSVMVLDGYEAITIARSAAHQQTDRVNPYGLHLGNRLPA
                     HTTSAGKILLAYLDDHAQQEWLNQYPLQRLTKYTYTNHIDFLRLLSEIKEQGWCYSSEE
                     HELGVHALAVPIYGQQSRVVAALNIVSPTMRTTKEYLIQHILPLLQETARELRNIL"
     regulatory      1983..2237
                     /label=induction operon terminator region
                     /note="induction operon terminator region"
                     /regulatory_class="terminator"
     rep_origin      complement(2549..3094)
                     /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(3186..3998)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 ccaattattg aaggcctccc taacgggggg cctttttttg tttctggtct cccgcttaac
       61 gatcgttggc tgttttgttc gattatcgaa caaattattg aaatatcgaa caaaacctct
      121 aaactactgt ggcactgaat caaaaaatta taaaccatga tcagaagctg tcaccggatg
      181 tgctttccgg tctgatgagt ccgtgaggac gaaacagcct ctacaaataa ttttgtttaa
      241 tactagagaa agaggggaaa tactagatgg tgagcaaggg cgaggagctg ttcaccgggg
      301 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
      361 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccacag
      421 gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc ctgcaatgct
      481 tcgcccgcta ccccgaccac atgaagctgc acgacttctt caagtccgcc atgcccgaag
      541 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
      601 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
      661 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
      721 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
      781 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacacccca atcggcgacg
      841 gccccgtgct gctgcccgac aaccactacc ttagctacca gtccgccctg agcaaagacc
      901 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
      961 tcggcatgga cgagctgtac aagtaactcg gtaccaaatt ccagaaaaga ggcctcccga
     1021 aaggggggcc ttttttcgtt ttggtccaat ggcggcgcgc catcgaatgg tgcaaaacct
     1081 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggaggtg aatatgtggt
     1141 cgaacatgga tgacaagaaa gtgaaagagg agaatattct gcacaattcc accaacaaga
     1201 agatcatccg ccacgaagat tttgtagccg gcattagcaa agggatggcg attctggatt
     1261 cgtttggtac agatcgtcat cgcctcaata tcaccatggc cgcagagaaa accggtatga
     1321 cacgtgcagc agctcgtcgc cacctgctta ctctggagta tctgggctat ctggaaagtg
     1381 acggccacta cttctactta actcccaaaa tcctgaaatt cagtggttca tatttgggtg
     1441 gtgctcaatt gccgaaaatt tcccaaccac tgttgaactt gcttacgacc cagaccagcc
     1501 tgatttacag cgtgatggtg ttggatggct atgaagccat taccattgcg cgttctgccg
     1561 ctcatcagca aaccgaccgc gttaacccgt atggtttaca tctcgggaat cgcttaccag
     1621 cgcatacaac gtcagcgggc aaaatcctgt tagcgtattt ggatgaccat gcccagcaag
     1681 agtggctcaa tcagtaccct ctgcaacggc tcacgaaata cacgtatacc aaccacatcg
     1741 actttctgcg ccttttgagt gaaatcaagg aacagggttg gtgctatagt tcggaagaac
     1801 acgaactggg agtacacgcc cttgcggttc cgatttacgg acagcagtct cgcgtcgtag
     1861 cggcactgaa cattgtcagc ccgacaatgc ggaccacgaa agaatacctg attcagcata
     1921 ttctgccgtt actgcaagaa actgcgcgtg aattgcgcaa tatcctgtaa tgataaggat
     1981 cctaattggt aacgaatcag acaattgacg gctcgaggga gtagcatagg gtttgcagaa
     2041 tccctgcttc gtccatttga caggcacatt atgcatcgat gataagctgt caaacatgag
     2101 cagatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct
     2161 ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg
     2221 agcaaatatt ttatctgagg tgcttcctcg ctcactgact cgctgcacga ggcagacctc
     2281 agcgctagcg gagtgtatac tggcttacta tgttggcact gatgagggtg tcagtgaagt
     2341 gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc agcagaatat gtgatacagg
     2401 atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcggcgagcg
     2461 gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat acttaacagg
     2521 gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct gacaagcatc
     2581 acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg
     2641 cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg
     2701 tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag ttccgggtag
     2761 gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc
     2821 ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc accactggca
     2881 gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta
     2941 aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc ggttcaaaga
     3001 gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga
     3061 gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta aggggtctga
     3121 cgctcagtgg aacgaaaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3181 ccttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     3241 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     3301 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     3361 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     3421 actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc
     3481 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     3541 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     3601 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     3661 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     3721 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     3781 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     3841 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     3901 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     3961 ggcctcgagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     4021 atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtacatc
     4081 agagattttg agacacaa
//
