LOCUS       Exported                5840 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA428, complete sequence.
ACCESSION   JX560390
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5840)
  AUTHORS   Silva-Rocha R, Martinez-Garcia E, Chavarria M, Calles B, 
            Arce-Rodriguez A, de las Heras A, Paez-Espino D, Durante-Rodriguez 
            G, Kim J, Nikel PI, Platero R, de Lorenzo V.
  TITLE     The Standard European Vector Architecture (SEVA): a coherent 
            platform for analysis and deployment of complex prokaryotic 
            phenotypes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5840)
  AUTHORS   Silva-Rocha R, Martinez-Garcia E, Chavarria M, Calles B, 
            Arce-Rodriguez A, de las Heras A, Paez-Espino D, Durante-Rodriguez 
            G, Kim J, Nikel PI, Platero R, de Lorenzo V.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-AUG-2012) Systems Biology Program, Centro Nacional de 
            Biotecnologia-CSIC, C/ Darwin 3, Campus de Cantoblanco, Madrid, 
            Madrid 28049, Spain
REFERENCE   3  (bases 1 to 5840)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: ApE v. v2.0.40
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5840
                     /organism="Cloning vector pSEVA428"
                     /mol_type="other DNA"
                     /db_xref="taxon:1246741"
     CDS             complement(19..984)
                     /codon_start=1
                     /gene="xylS"
                     /product="XylS"
                     /label=xylS
                     /protein_id="AFV59931.1"
                     /translation="MDFCLLNEKSQIFVHAEPYAVSDYVNQYVGTHSIRLPKGGRPAGR
                     LHHRIFGCLDLCRISYGGSVRVISPGLETCYHLQIILKGHCLWRGHGQEHYFAPGELLL
                     LNPDDQADLTYSEDCEKFIVKLPSVVLDRACSDNNWHKPREGIRFAARHNLQQLDGFIN
                     LLGLVCDEAEHTKSMPRVQEHYAGIIASKLLEMLGSNVSREIFSKGNPSFERVVQFIEE
                     NLKRNISLERLAELAMMSPRSLYNLFEKHAGTTPKNYIRNRKLESIRACLNDPSANVRS
                     ITEIALDYGFLHLGRFAENYRSAFGELPSDTLRQCKKEVA"
     gene            complement(19..984)
                     /gene="xylS"
                     /label=xylS
     promoter        1884..1964
                     /label=Pm promoter
                     /note="The bacterial Pm promoter is activated by XylS in 
                     the presence of benzoate or m-toluate (Marques et al., 
                     1999)."
     regulatory      1925..1977
                     /regulatory_class="promoter"
                     /note="Pm"
     misc_feature    2014..2070
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2079..2102)
                     /label=R24
                     /note="R24"
     regulatory      2109..2211
                     /regulatory_class="terminator"
                     /note="T0"
     terminator      2113..2207
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2329..3234
                     /codon_start=1
                     /gene="aadA"
                     /product="spectinomycin adenyltransferase"
                     /label=aadA
                     /protein_id="AFV59932.1"
                     /translation="MPRASKQQARYAVGRCLMLWSSNDVTQQGSRPKTKLNIMREAVIA
                     EVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRAL
                     INDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPA
                     TIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLFEALNETLTLWNSPPDWAGDER
                     NVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQPVILEARQAYLGQEEDRLASRA
                     DQLEEFVHYVKGEITKVVGK"
     gene            2329..3234
                     /gene="aadA"
                     /label=aadA
     CDS             2443..3234
                     /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"
     misc_feature    3255..3490
                     /label=oriT
                     /note="oriT"
     oriT            3380..3488
                     /note="incP origin of transfer"
     CDS             complement(3549..4697)
                     /codon_start=1
                     /gene="trfA"
                     /product="replication protein"
                     /label=trfA
                     /protein_id="AFV59933.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            complement(3549..4697)
                     /gene="trfA"
                     /label=trfA
     rep_origin      5090..5621
                     /label=oriV
                     /note="incP origin of replication"
     regulatory      5735..5839
                     /regulatory_class="terminator"
                     /note="T1"
     terminator      5748..5834
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"