LOCUS       Exported                3852 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAJM.661, complete sequence.
ACCESSION   MH101735
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3852)
  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 3852)
  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 3852)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3852
                     /organism="Cloning vector pAJM.661"
                     /mol_type="other DNA"
                     /db_xref="taxon:2461530"
     regulatory      1..53
                     /regulatory_class="terminator"
                     /note="L3S3P21"
     regulatory      73..123
                     /regulatory_class="promoter"
                     /note="PTtg"
     misc_feature    83..110
                     /label=TtgO operator
                     /note="TtgO operator"
     regulatory      85..90
                     /regulatory_class="minus_35_signal"
     regulatory      109..114
                     /regulatory_class="minus_10_signal"
     regulatory      121
                     /regulatory_class="other"
                     /note="+1"
     misc_feature    124..198
                     /label=RiboJ ribozyme insulator
                     /note="RiboJ ribozyme insulator"
     misc_RNA        126..175
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      207..218
                     /regulatory_class="ribosome_binding_site"
                     /note="B0064"
     CDS             225..944
                     /codon_start=1
                     /product="YFP"
                     /label=YFP
                     /protein_id="AYJ72251.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      945..1005
                     /regulatory_class="terminator"
                     /note="L3S2P21"
     regulatory      1010..1063
                     /regulatory_class="promoter"
                     /note="PLacIQ"
     regulatory      1028..1033
                     /regulatory_class="minus_35_signal"
     regulatory      1051..1056
                     /regulatory_class="minus_10_signal"
     regulatory      1064..1091
                     /regulatory_class="ribosome_binding_site"
                     /note="ttg2"
     CDS             1092..1724
                     /codon_start=1
                     /product="TtgR-AM"
                     /label=TtgR-AM
                     /protein_id="AYJ72252.1"
                     /translation="MVRRTKEEAQETRAQIIEAAERAFYKRGVARTTLADIAELAGVTR
                     GAIYWHFNNKAELVQALLDSLHETHDHLARASESEDEVDPLGCMRKLLLQVFNELVLDA
                     RTRRINEILHHKCEFTDDMCEIRQQHQSAVLDCHKGITLTLANVVRRGQLPGELDAERA
                     AVAMFAYVDGLIRRWLLLPDSVDLLGDVEKWVDTGLDMLRLSPALRK"
     regulatory      1737..1991
                     /regulatory_class="terminator"
                     /note="induction operon terminator region"
     rep_origin      2029..2865
                     /label=p15A origin
                     /note="p15A origin"
     rep_origin      complement(2303..2848)
                     /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(2937..3752)
                     /codon_start=1
                     /product="kanamycin phosphotransferase"
                     /label=kanamycin phosphotransferase
                     /protein_id="AYJ72253.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"