LOCUS       40924_6247        6373 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Integrational vector pBEXG, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6373)
  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 6373)
  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 6373)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6373)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Biotechnol."; date: "2013"; volume: "163"; issue: "2"; pages: 
            "143-154"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-FEB-2012) Systems Biology Program, Centro Nacional de 
            Biotecnologia (CNB-CSIC), C/Darwin 3, Madrid, Madrid 28049, Spain"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..19
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     CDS             complement(41..1003)
                     /codon_start=1
                     /label=XylS
                     /note="XylS regulator encoded by the Pseudomonas putida TOL
                     plasmid pWWO"
                     /translation="MDFCLLNEKSQIFVHAEPYAVSDYVNQYVGTHSIRLPKGGRPAGR
                     LHHRIFGCLDLCRISYGGSVRVISPGLETCYHLQIILKGHCLWRGHGQEHYFAPGELLL
                     LNPDDQADLTYSEDCEKFIVKLPSVVLDRACSDNNWHKPREGIRFAARHNLQQLDGFIN
                     LLGLVCDEAEHTKSMPRVQEHYAGIIASKLLEMLGSNVSREIFSKGNPSFERVVQFIEE
                     NLKRNISLERLAELAMMSPRSLYNLFEKHAGTTPKNYIRNRKLESIRACLNDPSANVRS
                     ITEIALDYGFLHLGRFAENYRSAFGELPSDTLRQCKKEVA"
     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)."
     misc_feature    2033..2089
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      2094..2188
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     protein_bind    2199..2246
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(2304..2834)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(3023..3051)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     misc_feature    3052..3099
                     /label=FRT
                     /note="FRT"
     protein_bind    3052..3099
                     /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(3106..3124)
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     CDS             3218..4645
                     /codon_start=1
                     /label=Tn5 transposase
                     /note="transposase from the bacterial Tn5 transposon
                     (Reznikoff, 1993)"
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     promoter        4702..4793
                     /label=AmpR promoter
     CDS             4794..5651
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      5658..5697
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            5844..5952
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      5971..6359
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 ctgtctctta tacacatctt taattaaacg ttcgtaatca agccacttcc tttttgcatt
       61 gacgcagggt gtcggaaggc aactcgccga acgcgctcct atagttttca gcgaagcgtc
      121 ccaaatgtaa gaagccgtag tctagggcta tctcagttat actacgcaca ttggcactgg
      181 gatcgttcaa gcaggcgcgg atgctttcga gcttgcggtt gcggatgtag ttcttcggcg
      241 tggtgccggc gtgcttctcg aacaaattgt agagcgagcg tggactcatc atcgccagct
      301 ccgctaaccg ctcaaggctg atattccgtt tgagattctc ctcaatgaat tgaacgactc
      361 gctcgaaaga cgggttacct ttgctgaaaa tttcacggct gacattgctg cccagcattt
      421 cgagcagctt ggaagcgatg atccccgcat agtgctcttg gacccgaggc atcgactttg
      481 tatgttccgc ttcgtcacaa actaacccga gtagattgat aaagccatcg agttgctgga
      541 gattgtgtcg cgcggcgaaa cggataccct ccctcggctt gtgccaattg ttgtcactgc
      601 acgcccgatc aaggaccact gagggcaatt taacgataaa tttctcgcaa tcttctgaat
      661 aggtcaggtc ggcttggtca tccggattga gcagcaatag ttcgcccggc gcaaaatagt
      721 gctcctggcc atggccacgc cacaggcaat ggcctttgag tattatttgc agatgataac
      781 aggtctctaa tccaggcgag attaccctca cgctaccgcc gtagctgatt cgacacagat
      841 cgaggcatcc gaagattctg tggtgcagcc tgcctgccgg gcgcccgccc ttgggcaggc
      901 gaatagagtg cgtaccgaca tactggttaa cataatcgga gactgcatag ggctcggcgt
      961 ggacgaagat ctgacttttc tcgttcaata agcaaaaatc catagttcac ggttctctta
     1021 ttttaatgtg ggctgcttgg tgtgatgtag aaaggcgcca agtcgatgaa aatgcatctc
     1081 gacgtgatgc gtatacgggt tacccccatt gccacgttgc gccatccttt ttgcaatcag
     1141 tgaccacttt tccaagcaaa aataacgcca agcagaacga agacgttctt tttaagaagc
     1201 gagaacacca gaagttcgtg ctgtcggggc atggggcgac gaattggcgg ataaagggga
     1261 tctgctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
     1321 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccgtaattac gtatggcaat
     1381 gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga
     1441 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct
     1501 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg
     1561 gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga
     1621 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgc atgggaatta
     1681 gcttgatctg accaacgacc ggtagcggag ctatccaacg gcggtatacc aggaaaacac
     1741 acagcaggta catcagaaca gtaccatgac tgaagaacaa atagtttttt cctgatccat
     1801 aaagcagaac ggcctgctcc atgacaaatc tggctcccca actaatgccc catgcagcca
     1861 gcataaccag cataaagtgc agtgtccggt ttgataggga taagtccagc cttgcaagaa
     1921 gcggatacag gagtgcaaaa aatggctatc tctagtaagg cctacccctt aggctttatg
     1981 caacagaaac aataataatg gagtcatgac catgcctagg ccgcggccgc gcgaattcga
     2041 gctcggtacc cggggatcct ctagagtcga cctgcaggca tgcaagcttc ttggactcct
     2101 gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg
     2161 ccgccgggcg ttttttattg gtgagaatcc agactagtga agttcctatt ccgaagttcc
     2221 tattctctag aaagtatagg aacttccaat ttaccgaaca actccgcggc cgggaagccg
     2281 atctcggctt gaacgaattg ttaggtggcg gtacttgggt cgatatcaaa gtgcatcact
     2341 tcttcccgta tgcccaactt tgtatagaga gccactgcgg gatcgtcacc gtaatctgct
     2401 tgcacgtaga tcacataagc accaagcgcg ttggcctcat gcttgaggag attgatgagc
     2461 gcggtggcaa tgccctgcct ccggtgctct ccggagactg cgagatcata gatatagatc
     2521 tcactacgcg gctgctcaaa cttgggcaga acgtaagccg cgagagcgcc aacaaccgct
     2581 tcttggtcga aggcagcaag cgcgatgaat gtcttactac ggagcaagtt cccgaggtaa
     2641 tcggagtccg gctgatgttg ggagtaggtg gctacgtctc cgaactcacg accgaaaaga
     2701 tcaagagcag cccgcatgga tttgacttgg tcagggccga gcctacatgt gcgaatgatg
     2761 cccatacttg agccacctaa ctttgtttta gggcgactgc cctgctgcgt aacatcgttg
     2821 ctgctgcgta acatcgttgc tgctccataa catcaaacat cgacccacgg cgtaacgcgc
     2881 ttgctgcttg gatgcccgag gcataggctg tacaaaaaaa cagtcataac aagccatgaa
     2941 aaccgccact gcgccgttac caccgctgcg ttcggtcaag gttctggacc agttgcgtga
     3001 gcgcatacgc tacttgcatt acagtttacg aaccgaacag gcttatgtca agaagttcct
     3061 attccgaagt tcctattctc tagaaagtat aggaacttcg acgtcagatg tgtataagag
     3121 acagctggtt taaactacac aagtagcgtc ctgaacggaa cctttcccgt tttccagaat
     3181 ctgatgttcc atgtgacctc ctaacatggt aacgttcatg attaccagtg cactgcatcg
     3241 tgcggcggat tgggcgaaaa gcgtgttttc tagtgctgcg ctgggtgatc cgcgtcgtac
     3301 cgcgcgtctg gtgaatgttg cggcgcaact ggccaaatat agcggcaaaa gcattaccat
     3361 tagcagcgaa ggcagcaaag ccatgcagga aggcgcgtat cgttttattc gtaatccgaa
     3421 cgtgagcgcg gaagcgattc gtaaagcggg tgccatgcag accgtgaaac tggcccagga
     3481 atttccggaa ctgctggcaa ttgaagatac cacctctctg agctatcgtc atcaggtggc
     3541 ggaagaactg ggcaaactgg gtagcattca ggataaaagc cgtggttggt gggtgcatag
     3601 cgtgctgctg ctggaagcga ccacctttcg taccgtgggc ctgctgcatc aagaatggtg
     3661 gatgcgtccg gatgatccgg cggatgcgga tgaaaaagaa agcggcaaat ggctggccgc
     3721 tgctgcaact tcgcgtctga gaatgggcag catgatgagc aacgtgattg cggtgtgcga
     3781 tcgtgaagcg gatattcatg cgtatctgca agataaactg gcccataacg aacgttttgt
     3841 ggtgcgtagc aaacatccgc gtaaagatgt ggaaagcggc ctgtatctgt atgatcacct
     3901 gaaaaaccag ccggaactgg gcggctatca gattagcatt ccgcagaaag gcgtggtgga
     3961 taaacgtggc aaacgtaaaa accgtccggc gcgtaaagcg agcctgagcc tgcgtagcgg
     4021 ccgtattacc ctgaaacagg gcaacattac cctgaacgcg gtgctggccg aagaaattaa
     4081 tccgccgaaa ggcgaaaccc cgctgaaatg gctgctgctg accagcgagc cggtggaaag
     4141 tctggcccaa gcgctgcgtg tgattgatat ttatacccat cgttggcgca ttgaagaatt
     4201 tcacaaagcg tggaaaacgg gtgcgggtgc ggaacgtcag cgtatggaag aaccggataa
     4261 cctggaacgt atggtgagca ttctgagctt tgtggcggtg cgtctgctgc aactgcgtga
     4321 atcttttact ccgccgcaag cactgcgtgc gcagggcctg ctgaaagaag cggaacacgt
     4381 tgaaagccag agcgcggaaa ccgtgctgac cccggatgaa tgccaactgc tgggctatct
     4441 ggataaaggc aaacgcaaac gcaaagaaaa agcgggcagc ctgcaatggg cgtatatggc
     4501 gattgcgcgt ctgggcggct ttatggatag caaacgtacc ggcattgcga gctggggtgc
     4561 gctgtgggaa ggttgggaag cgctgcaaag caaactggat ggctttctgg ccgcgaaaga
     4621 cctgatggcg cagggcatta aaatctaatc gatttaaatt agtagcccgc ctaatgagcg
     4681 ggcttttttt taattcccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4741 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagca
     4801 ttcagcattt tcgtgtggcg ctgattccgt tttttgcggc gttttgcctg ccggtgtttg
     4861 cgcatccgga aaccctggtg aaagtgaaag atgcggaaga tcaactgggt gcgcgcgtgg
     4921 gctatattga actggatctg aacagcggca aaattctgga atcttttcgt ccggaagaac
     4981 gttttccgat gatgagcacc tttaaagtgc tgctgtgcgg tgcggttctg agccgtgtgg
     5041 atgcgggcca ggaacaactg ggccgtcgta ttcattatag ccagaacgat ctggtggaat
     5101 atagcccggt gaccgaaaaa catctgaccg atggcatgac cgtgcgtgaa ctgtgcagcg
     5161 cggcgattac catgagcgat aacaccgcgg cgaacctgct gctgacgacc attggcggtc
     5221 cgaaagaact gaccgcgttt ctgcataaca tgggcgatca tgtgacccgt ctggatcgtt
     5281 gggaaccgga actgaacgaa gcgattccga acgatgaacg tgataccacc atgccggcag
     5341 caatggcgac caccctgcgt aaactgctga cgggtgagct gctgaccctg gcaagccgcc
     5401 agcaactgat tgattggatg gaagcggata aagtggcggg tccgctgctg cgtagcgcgc
     5461 tgccggctgg ctggtttatt gcggataaaa gcggtgcggg cgaacgtggc agccgtggca
     5521 ttattgcggc gctgggcccg gatggtaaac cgagccgtat tgtggtgatt tataccaccg
     5581 gcagccaggc gacgatggat gaacgtaacc gtcagattgc ggaaattggc gcgagcctga
     5641 ttaaacattg gtaaaccgat acaattaaag gctccttttg gagccttttt ttttggacga
     5701 cccttgtcct tttccgctgc ataaccctgc ttcggggtca ttatagcgat tttttcggta
     5761 tatccatcct ttttcgcacg atatacagga ttttgccaaa gggttcgtgt agactttcct
     5821 tggtgtatcc aacggcgtca gccgggcagg ataggtgaag taggcccacc cgcgagcggg
     5881 tgttccttct tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg
     5941 cgaggctggc cgtaggccgg ccggcgcgcc gatctgaaga tcagcagttc aacctgttga
     6001 tagtacgtac taagctctca tgtttcacgt actaagctct catgtttaac gtactaagct
     6061 ctcatgttta acgaactaaa ccctcatggc taacgtacta agctctcatg gctaacgtac
     6121 taagctctca tgtttcacgt actaagctct catgtttgaa caataaaatt aatataaatc
     6181 agcaacttaa atagcctcta aggttttaag ttttataaga aaaaaaagaa tatataaggc
     6241 ttttaaagcc tttaaggttt aacggttgtg gacaacaagc cagggatgta acgcactgag
     6301 aagcccttag agcctctcaa agcaattttg agtgacacag gaacacttaa cggctgacat
     6361 ggggcgcgcc cag
//
