LOCUS       Exported                6118 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA528, complete sequence.
ACCESSION   JX560391
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6118)
  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  JOURNAL   Unpublished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 6118)
  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 6118)
  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..6118
                     /organism="Cloning vector pSEVA528"
                     /mol_type="other DNA"
                     /db_xref="taxon:1246753"
     CDS             complement(19..984)
                     /codon_start=1
                     /gene="xylS"
                     /product="XylS"
                     /label=xylS
                     /protein_id="AFV59934.1"
                     /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             2322..3512
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     CDS             2394..3512
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux transporter"
                     /label=tetA
                     /protein_id="AFV59935.1"
                     /translation="MPVLPGLLRDIVHSDSIASHYGVLLALYALMQFLCAPVLGALSDR
                     FGRRPVLLASLLGATIDYAIMATTPVLWILYAGRIVAGITGATGAVAGAYIADITDGED
                     RARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCFLMQESHKG
                     ERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVIFGEDRFRWSAT
                     MIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVLLAFATRGWMAFP
                     IMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITGPLIVTAIYAASAS
                     TWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            2394..3512
                     /gene="tetA"
                     /label=tetA
     misc_feature    3533..3768
                     /label=oriT
                     /note="oriT"
     oriT            3658..3766
                     /note="incP origin of transfer"
     CDS             complement(3827..4975)
                     /codon_start=1
                     /gene="trfA"
                     /product="replication protein"
                     /label=trfA
                     /protein_id="AFV59936.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            complement(3827..4975)
                     /gene="trfA"
                     /label=trfA
     rep_origin      5368..5899
                     /label=oriV
                     /note="incP origin of replication"
     regulatory      6013..6117
                     /regulatory_class="terminator"
                     /note="T1"
     terminator      6026..6112
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"