LOCUS       Exported                8180 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFM141, complete sequence.
ACCESSION   KF322085
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8180)
  AUTHORS   Moser F, Horwitz A, Chen J, Lim W, Voigt CA.
  TITLE     Genetic sensor for strong methylating compounds
  JOURNAL   ACS Synth Biol 2 (10), 614-624 (2013)
  PUBMED    24032656
REFERENCE   2  (bases 1 to 8180)
  AUTHORS   Moser F, Horwitz A, Chen J, Lim WA, Voigt CA.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-JUL-2013) Biological Engineering, MIT, 500 Tech 
            Square, Rm 209D, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 8180)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8180
                     /organism="Cloning vector pFM141"
                     /mol_type="other DNA"
                     /db_xref="taxon:1403156"
     CDS             62..853
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     CDS             74..853
                     /codon_start=1
                     /product="StrepR"
                     /label=StrepR
                     /protein_id="AGW27644.1"
                     /translation="MIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPHSDID
                     LLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAKRELQ
                     FGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLFEALN
                     ETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQPVILE
                     ARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     terminator      1130..1159
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     regulatory      1261..1338
                     /regulatory_class="terminator"
                     /note="terminator B0055"
     terminator      1288..1317
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    1364..1385
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(1386..2264)
                     /codon_start=1
                     /product="AraC"
                     /label=AraC
                     /protein_id="AGW27646.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2291..2575
                     /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)"
     regulatory      2292..2595
                     /regulatory_class="promoter"
                     /note="promoter PBAD"
     misc_feature    2604..2616
                     /label=RBS B0032
                     /note="RBS B0032"
     CDS             2623..3687
                     /codon_start=1
                     /product="Ada"
                     /label=Ada
                     /protein_id="AGW27645.1"
                     /translation="MKKATCLTDDQRWQSVLARDPNADGEFVFAVRTTGIFCRPSCRAR
                     HALRENVSFYANASEALAAGFRPCKRCQPEKANAQQHRLDKITHACRLLEQETPVTLEA
                     LADQVAMSPFHLHRLFKATTGMTPKAWQQAWRARRLRESLAKGESVTTSILNAGFPDSS
                     SYYRKADETLGMTAKQFRHGGENLAVRYALADCELGRCLVAESERGICAILLGDDDATL
                     ISELQQMFPAADNAPADLMFQQHVREVIASLNQRDTPLTLPLDIRGTAFQQQVWQALRT
                     IPCGETVSYQQLANAIGKPKAVRAVASACAANKLAIIIPCHRVVRGDGTLSGYRWGVSR
                     KAQLLRREAENEER"
     misc_feature    3688..3708
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     regulatory      3734..3802
                     /regulatory_class="terminator"
                     /note="terminator B0054"
     terminator      3744..3775
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             4662..5633
                     /codon_start=1
                     /product="RepA"
                     /label=RepA
                     /protein_id="AGW27647.1"
                     /translation="MPKNNKAPGHRINEIIKTSLALEMEDAREAGLVGYMARCLVQATM
                     PHTDPKTSYFERTNGIVTLSIMGKPSIGLPYGSMPRTLLAWICTEAVRTKDPVLNLGRS
                     QSEFLQRLGMHTDGRYTATLRNQAQRLFSSMISLAGEQGNDFGIENVVIAKRAFLFWNP
                     KRPEDRALWDSTLTLTGDFFEEVTRSPVPIRIDYLHALRQSPLAMDIYTWLTYRVFLLR
                     AKGRPFVQIPWVALQAQFGSSYGSRARNSPELDDKARERAERAALASFKYNFKKRLREV
                     LIVYPEASDCIEDDGECLRIKSTRLHVTRAPGKGARIGPPPT"
     rep_origin      5670..6105
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     rep_origin      7634..8013
                     /label=R6K origin of replication
                     /note="R6K origin of replication"
     rep_origin      7638..8026
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"