LOCUS       Exported                5793 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAJM.677, complete sequence.
ACCESSION   MH101733
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5793)
  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 5793)
  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 5793)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5793
                     /organism="Cloning vector pAJM.677"
                     /mol_type="other DNA"
                     /db_xref="taxon:2461531"
     regulatory      1..53
                     /regulatory_class="terminator"
                     /note="L3S3P21"
     regulatory      73..361
                     /regulatory_class="promoter"
                     /note="PBAD"
     promoter        73..356
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     misc_feature    74..90
                     /label=O2 operator
                     /note="O2 operator"
     misc_feature    211..228
                     /label=O1L operator
                     /note="O1L operator"
     misc_feature    231..248
                     /label=O1R operator
                     /note="O1R operator"
     misc_feature    249..281
                     /label=CAP operator
                     /note="CAP operator"
     misc_feature    285..300
                     /label=I1 operator
                     /note="I1 operator"
     misc_feature    306..317
                     /label=I2 operator
                     /note="I2 operator"
     regulatory      319..324
                     /regulatory_class="minus_35_signal"
     regulatory      341..346
                     /regulatory_class="minus_10_signal"
     regulatory      355
                     /regulatory_class="other"
                     /note="+1"
     misc_feature    362..436
                     /label=RiboJ ribozyme insulator
                     /note="RiboJ ribozyme insulator"
     misc_RNA        364..413
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      445..456
                     /regulatory_class="ribosome_binding_site"
                     /note="B0064"
     CDS             463..1182
                     /codon_start=1
                     /product="YFP"
                     /label=YFP
                     /protein_id="AYJ72244.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      1183..1243
                     /regulatory_class="terminator"
                     /note="L3S2P21"
     regulatory      1248..1301
                     /regulatory_class="promoter"
                     /note="PLacIQ"
     regulatory      1266..1271
                     /regulatory_class="minus_35_signal"
     regulatory      1289..1294
                     /regulatory_class="minus_10_signal"
     regulatory      1302..1329
                     /regulatory_class="ribosome_binding_site"
                     /note="ara1"
     CDS             1330..2208
                     /codon_start=1
                     /product="AraC-AM"
                     /label=AraC-AM
                     /protein_id="AYJ72245.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDFFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MLAINGSLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGLE
                     EKVNDVAVKLS"
     regulatory      2209..2249
                     /regulatory_class="ribosome_binding_site"
                     /note="e1"
     CDS             2250..3668
                     /codon_start=1
                     /product="AraE"
                     /label=AraE
                     /protein_id="AYJ72246.1"
                     /translation="MVTINTESALTPRSLRDTRRMNMFVSVAAAVAGLLFGLDIGVIAG
                     ALPFITDHFVLTSRLQEWVVSSMMLGAAIGALFNGWLSFRLGRKYSLMAGAILFVLGSI
                     GSAFATSVEMLIAARVVLGIAVGIASYTAPLYLSEMASENVRGKMISMYQLMVTLGIVL
                     AFLSDTAFSYSGNWRAMLGVLALPAVLLIILVVFLPNSPRWLAEKGRHIEAEEVLRMLR
                     DTSEKAREELNEIRESLKLKQGGWALFKINRNVRRAVFLGMLLQAMQQFTGMNIIMYYA
                     PRIFKMAGFTTTEQQMIATLVVGLTFMFATFIAVFTVDKAGRKPALKIGFSVMALGTLV
                     LGYCLMQFDNGTASSGLSWLSVGMTMMCIAGYAMSAAPVVWILCSEIQPLKCRDFGITC
                     STTTNWVSNMIIGATFLTLLDSIGAAGTFWLYTALNIAFVGITFWLIPETKNVTLEHIE
                     RKLMAGEKLRNIGV"
     regulatory      3678..3932
                     /regulatory_class="terminator"
                     /note="induction operon terminator region"
     rep_origin      3970..4806
                     /label=p15A origin
                     /note="p15A origin"
     rep_origin      complement(4244..4789)
                     /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(4878..5693)
                     /codon_start=1
                     /product="kanamycin phosphotransferase"
                     /label=kanamycin phosphotransferase
                     /protein_id="AYJ72247.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"