LOCUS       Exported                3973 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAJM.474, complete sequence.
ACCESSION   MH101729
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3973)
  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 3973)
  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 3973)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3973
                     /organism="Cloning vector pAJM.474"
                     /mol_type="other DNA"
                     /db_xref="taxon:2461528"
     regulatory      1..53
                     /regulatory_class="terminator"
                     /note="L3S3P21"
     regulatory      73..127
                     /regulatory_class="promoter"
                     /note="PLuxB"
     misc_feature    73..92
                     /label=LuxO operator
                     /note="LuxO operator"
     regulatory      93..98
                     /regulatory_class="minus_35_signal"
     regulatory      114..119
                     /regulatory_class="minus_10_signal"
     misc_feature    128..202
                     /label=RiboJ ribozyme insulator
                     /note="RiboJ ribozyme insulator"
     misc_RNA        130..179
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      211..222
                     /regulatory_class="ribosome_binding_site"
                     /note="B0064"
     CDS             229..948
                     /codon_start=1
                     /product="YFP"
                     /label=YFP
                     /protein_id="AYJ72232.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      949..1009
                     /regulatory_class="terminator"
                     /note="L3S2P21"
     regulatory      1014..1067
                     /regulatory_class="promoter"
                     /note="PLacI"
     regulatory      1032..1037
                     /regulatory_class="minus_35_signal"
     regulatory      1055..1060
                     /regulatory_class="minus_10_signal"
     regulatory      1068..1095
                     /regulatory_class="ribosome_binding_site"
                     /note="lux1"
     CDS             1096..1848
                     /codon_start=1
                     /product="LuxR"
                     /label=LuxR
                     /protein_id="AYJ72233.1"
                     /translation="MKNINADDTYRIINKIKACRSNNDINQCLSDMTKMVHCEYYLLAI
                     IYPHSMVKSDISILDNYPKKWRQYYDDANLIKYDPIVDYSNSNHSPINWNIFENNAVNK
                     KSPNVIKEAKTSGLITGFSFPIHTANNGFGMLSFAHSEKDNYIDSLFLHACMNIPLIVP
                     SLVDNYRKINIANNKSNNDLTKREKECLAWACEGKSSWDISKILGCSERTVTFHLTNAQ
                     MKLNTTNRCQSISKAILTGAIDCPYFKN"
     regulatory      1858..2112
                     /regulatory_class="terminator"
                     /note="induction operon terminator region"
     rep_origin      2150..2986
                     /label=p15A origin
                     /note="p15A origin"
     rep_origin      complement(2424..2969)
                     /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(3058..3873)
                     /codon_start=1
                     /product="kanamycin phosphotransferase"
                     /label=kanamycin phosphotransferase
                     /protein_id="AYJ72234.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"