LOCUS       Exported                4558 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAJM.969, complete sequence.
ACCESSION   MH101740
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4558)
  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 4558)
  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 4558)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4558
                     /organism="Cloning vector pAJM.969"
                     /mol_type="other DNA"
                     /db_xref="taxon:2461549"
     regulatory      1..53
                     /regulatory_class="terminator"
                     /note="L3S3P21"
     regulatory      73..124
                     /regulatory_class="promoter"
                     /note="PMph"
     misc_feature    74..108
                     /label=MphO operator
                     /note="MphO operator"
     regulatory      76..81
                     /regulatory_class="minus_35_signal"
     regulatory      99..104
                     /regulatory_class="minus_10_signal"
     regulatory      111
                     /regulatory_class="other"
                     /note="+1"
     misc_feature    125..199
                     /label=RiboJ ribozyme insulator
                     /note="RiboJ ribozyme insulator"
     misc_RNA        127..176
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      208..219
                     /regulatory_class="ribosome_binding_site"
                     /note="B0064"
     CDS             226..945
                     /codon_start=1
                     /product="YFP"
                     /label=YFP
                     /protein_id="AYJ72266.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      946..1006
                     /regulatory_class="terminator"
                     /note="L3S2P21"
     regulatory      1011..1064
                     /regulatory_class="promoter"
                     /note="PLacIQ"
     regulatory      1029..1034
                     /regulatory_class="minus_35_signal"
     regulatory      1052..1057
                     /regulatory_class="minus_10_signal"
     regulatory      1065..1092
                     /regulatory_class="ribosome_binding_site"
                     /note="mph2"
     CDS             1093..1677
                     /codon_start=1
                     /product="MphR-AM"
                     /label=MphR-AM
                     /protein_id="AYJ72267.1"
                     /translation="MPRPKLKSDDEVLEAATVVLKRCGPIEFTLSGVAKEVGLSRAALI
                     QRFTNRDTLLVRMMERGVEQVRHYLNAIPIGAGPQGLWEFLQVLVRSMDTRNDFSVNYL
                     ISWYELQVPELRTLAIQRNRAVVEGIRKRLPPGAPAAAELLLHSVIAGATMQWAVDPDG
                     ELADHVLAQIAAILCLMFPEHDDFQLLQAHA"
     regulatory      1678..1698
                     /regulatory_class="ribosome_binding_site"
                     /note="ery1"
     CDS             1699..2433
                     /codon_start=1
                     /product="EryR"
                     /label=EryR
                     /protein_id="AYJ72268.1"
                     /translation="MNEKNIKHSQNFITSKHNIDKIMTNIRLNEHDNIFEIGSGKGHFT
                     LELVKRCNFVTAIEIDHKLCKTTENKLVDHDNFQVLNKDILQFKFPKNQSYKIYGNIPY
                     NISTDIIRKIVFDSIANEIYLIVEYGFAKRLLNTKRSLALLLMAEVDISILSMVPREYF
                     HPKPKVNSSLIRLSRKKSRISHKDKQKYNYFVMKWVNKEYKKIFTKNQFNNSLKHAGID
                     DLNNISFEQFLSLFNSYKLFNK"
     regulatory      2443..2697
                     /regulatory_class="terminator"
                     /note="induction operon terminator region"
     rep_origin      2735..3571
                     /label=p15A origin
                     /note="p15A origin"
     rep_origin      complement(3009..3554)
                     /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(3643..4458)
                     /codon_start=1
                     /product="kanamycin phosphotransferase"
                     /label=kanamycin phosphotransferase
                     /protein_id="AYJ72269.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"