LOCUS       Exported                3867 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAJM.657, complete sequence.
ACCESSION   MH101728
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3867)
  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 3867)
  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 3867)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3867
                     /organism="Cloning vector pAJM.657"
                     /mol_type="other DNA"
                     /db_xref="taxon:2461529"
     regulatory      1..53
                     /regulatory_class="terminator"
                     /note="L3S3P21"
     regulatory      73..162
                     /regulatory_class="promoter"
                     /note="PCymRC"
     misc_feature    73..104
                     /label=CymO operator
                     /note="CymO operator"
     protein_bind    73..100
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     regulatory      96..101
                     /regulatory_class="minus_35_signal"
     regulatory      119..124
                     /regulatory_class="minus_10_signal"
     misc_feature    131..162
                     /label=CymO operator
                     /note="CymO operator"
     protein_bind    131..158
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     regulatory      131
                     /regulatory_class="other"
                     /note="+1"
     misc_feature    163..237
                     /label=RiboJ ribozyme insulator
                     /note="RiboJ ribozyme insulator"
     misc_RNA        165..214
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      246..257
                     /regulatory_class="ribosome_binding_site"
                     /note="B0064"
     CDS             264..983
                     /codon_start=1
                     /product="YFP"
                     /label=YFP
                     /protein_id="AYJ72229.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      984..1044
                     /regulatory_class="terminator"
                     /note="L3S2P21"
     regulatory      1049..1102
                     /regulatory_class="promoter"
                     /note="PLacIQ"
     regulatory      1067..1072
                     /regulatory_class="minus_35_signal"
     regulatory      1090..1095
                     /regulatory_class="minus_10_signal"
     regulatory      1103..1130
                     /regulatory_class="ribosome_binding_site"
                     /note="cym1"
     CDS             1131..1742
                     /codon_start=1
                     /product="CymR-AM"
                     /label=CymR-AM
                     /protein_id="AYJ72230.1"
                     /translation="MSPKRRTQAERAMETQGKLIAAALGVLREKGYAGFRIADVPGAAG
                     VSRGAQSHHFPTKLELLLATFEWLYEQITERSRARLAKLKPEDDVIQQMLDDAAEFFLD
                     DDFSIGLDLIVAADRDPALREGIQRTVERNRFVVEDMWLGVLVSRGLSRDDAEDILWLI
                     FNSVRGLVVRSLWQKDKERFERVRNSTLEIARERYAKFKR"
     regulatory      1752..2006
                     /regulatory_class="terminator"
                     /note="induction operon terminator region"
     rep_origin      2044..2879
                     /label=p15A origin
                     /note="p15A origin"
     rep_origin      complement(2318..2863)
                     /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(2952..3767)
                     /codon_start=1
                     /product="kanamycin phosphotransferase"
                     /label=kanamycin phosphotransferase
                     /protein_id="AYJ72231.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"