LOCUS       Exported                6495 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Integrational vector pBEXK, complete sequence.
ACCESSION   JQ663866
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6495)
  AUTHORS   Nikel PI, de Lorenzo V.
  TITLE     Implantation of unmarked regulatory and metabolic modules in 
            Gram-negative bacteria with specialised mini-transposon delivery 
            vectors
  JOURNAL   J. Biotechnol. 163 (2), 143-154 (2013)
  PUBMED    22609234
REFERENCE   2  (bases 1 to 6495)
  AUTHORS   Nikel PI, de Lorenzo V.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-FEB-2012) Systems Biology Program, Centro Nacional de 
            Biotecnologia (CNB-CSIC), C/Darwin 3, Madrid, Madrid 28049, Spain
REFERENCE   3  (bases 1 to 6495)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6495
                     /organism="Integrational vector pBEXK"
                     /mol_type="other DNA"
                     /db_xref="taxon:1219768"
     misc_feature    complement(1..19)
                     /note="ME-O; mosaic element O"
     CDS             complement(38..1003)
                     /codon_start=1
                     /gene="xylS"
                     /product="XylS regulator"
                     /label=xylS
                     /protein_id="AFZ99883.1"
                     /translation="MDFCLLNEKSQIFVHAEPYAVSDYVNQYVGTHSIRLPKGGRPAGR
                     LHHRIFGCLDLCRISYGGSVRVISPGLETCYHLQIILKGHCLWRGHGQEHYFAPGELLL
                     LNPDDQADLTYSEDCEKFIVKLPSVVLDRACSDNNWHKPREGIRFAARHNLQQLDGFIN
                     LLGLVCDEAEHTKSMPRVQEHYAGIIASKLLEMLGSNVSREIFSKGNPSFERVVQFIEE
                     NLKRNISLERLAELAMMSPRSLYNLFEKHAGTTPKNYIRNRKLESIRACLNDPSANVRS
                     ITEIALDYGFLHLGRFAENYRSAFGELPSDTLRQCKKEVA"
     gene            complement(38..1003)
                     /gene="xylS"
                     /label=xylS
     promoter        1903..1983
                     /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      1944..1996
                     /regulatory_class="promoter"
     misc_feature    2033..2089
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     regulatory      2090..2192
                     /regulatory_class="terminator"
     terminator      2094..2188
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    2199..2246
                     /label=FRT
                     /note="FRT"
     protein_bind    2199..2246
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(2257..3072)
                     /codon_start=1
                     /gene="aphA"
                     /product="AphA"
                     /label=aphA
                     /note="aminoglycoside 3-phosphotransferase"
                     /protein_id="AFZ99880.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(2257..3072)
                     /gene="aphA"
                     /label=aphA
     misc_feature    3174..3221
                     /label=FRT
                     /note="FRT"
     protein_bind    3174..3221
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    complement(3228..3246)
                     /note="ME-I; mosaic element I"
     CDS             3340..4770
                     /codon_start=1
                     /gene="tnpA"
                     /product="Tn5 transposase"
                     /label=tnpA
                     /protein_id="AFZ99882.1"
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     gene            3340..4770
                     /gene="tnpA"
                     /label=tnpA
     CDS             3340..4767
                     /codon_start=1
                     /product="transposase from the bacterial Tn5 transposon 
                     (Reznikoff, 1993)"
                     /label=Tn5 transposase
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     promoter        4824..4915
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4916..5776
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AFZ99881.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            4916..5776
                     /gene="bla"
                     /label=bla
     terminator      5780..5819
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     misc_feature    complement(5831..6076)
                     /note="oriT; RK2"
     oriT            5966..6074
                     /note="incP origin of transfer"
     rep_origin      complement(6093..6484)
                     /direction=LEFT
     rep_origin      6093..6481
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"