LOCUS       40924_39558        6118 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA528, complete sequence.
ACCESSION   .
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  JOURNAL   Unpublished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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6118)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-AUG-2012) Systems Biology Program, Centro Nacional de 
            Biotecnologia-CSIC, C/ Darwin 3, Campus de Cantoblanco, Madrid, 
            Madrid 28049, Spain"
COMMENT     SGRef: number: 3; type: "Journal Article"
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
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(22..984)
                     /codon_start=1
                     /label=XylS
                     /note="XylS regulator encoded by the Pseudomonas putida TOL
                     plasmid pWWO"
                     /translation="MDFCLLNEKSQIFVHAEPYAVSDYVNQYVGTHSIRLPKGGRPAGR
                     LHHRIFGCLDLCRISYGGSVRVISPGLETCYHLQIILKGHCLWRGHGQEHYFAPGELLL
                     LNPDDQADLTYSEDCEKFIVKLPSVVLDRACSDNNWHKPREGIRFAARHNLQQLDGFIN
                     LLGLVCDEAEHTKSMPRVQEHYAGIIASKLLEMLGSNVSREIFSKGNPSFERVVQFIEE
                     NLKRNISLERLAELAMMSPRSLYNLFEKHAGTTPKNYIRNRKLESIRACLNDPSANVRS
                     ITEIALDYGFLHLGRFAENYRSAFGELPSDTLRQCKKEVA"
     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)."
     misc_feature    2014..2070
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2079..2102)
                     /label=R24
                     /note="R24"
     terminator      2113..2207
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2322..3509
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     oriT            3658..3766
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(3830..4975)
                     /codon_start=1
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(5368..5899)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(6026..6112)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ttaattaaac gttcgtaatc aagccacttc ctttttgcat tgacgcaggg tgtcggaagg
       61 caactcgccg aacgcgctcc tatagttttc agcgaagcgt cccaaatgta agaagccgta
      121 gtctagggct atctcagtta tactacgcac attggcactg ggatcgttca agcaggcgcg
      181 gatgctttcg agcttgcggt tgcggatgta gttcttcggc gtggtgccgg cgtgcttctc
      241 gaacaaattg tagagcgagc gtggactcat catcgccagc tccgctaacc gctcaaggct
      301 gatattccgt ttgagattct cctcaatgaa ttgaacgact cgctcgaaag acgggttacc
      361 tttgctgaaa atttcacggc tgacattgct gcccagcatt tcgagcagct tggaagcgat
      421 gatccccgca tagtgctctt ggacccgagg catcgacttt gtatgttccg cttcgtcaca
      481 aactaacccg agtagattga taaagccatc gagttgctgg agattgtgtc gcgcggcgaa
      541 acggataccc tccctcggct tgtgccaatt gttgtcactg cacgcccgat caaggaccac
      601 tgagggcaat ttaacgataa atttctcgca atcttctgaa taggtcaggt cggcttggtc
      661 atccggattg agcagcaata gttcgcccgg cgcaaaatag tgctcctggc catggccacg
      721 ccacaggcaa tggcctttga gtattatttg cagatgataa caggtctcta atccaggcga
      781 gattaccctc acgctaccgc cgtagctgat tcgacacaga tcgaggcatc cgaagattct
      841 gtggtgcagc ctgcctgccg ggcgcccgcc cttgggcagg cgaatagagt gcgtaccgac
      901 atactggtta acataatcgg agactgcata gggctcggcg tggacgaaga tctgactttt
      961 ctcgttcaat aagcaaaaat ccatagttca cggttctctt attttaatgt gggctgcttg
     1021 gtgtgatgta gaaaggcgcc aagtcgatga aaatgcatct cgacgtgatg cgtatacggg
     1081 ttacccccat tgccacgttg cgccatcctt tttgcaatca gtgaccactt ttccaagcaa
     1141 aaataacgcc aagcagaacg aagacgttct ttttaagaag cgagaacacc agaagttcgt
     1201 gctgtcgggg catggggcga cgaattggcg gataaagggg atctgctgga tattacggcc
     1261 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
     1321 gcccgcctga tgaatgctca tccgtaatta cgtatggcaa tgaaagacgg tgagctggtg
     1381 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
     1441 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
     1501 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
     1561 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
     1621 gacaacttct tcgcccccgt tttcaccatg catgggaatt agcttgatct gaccaacgac
     1681 cggtagcgga gctatccaac ggcggtatac caggaaaaca cacagcaggt acatcagaac
     1741 agtaccatga ctgaagaaca aatagttttt tcctgatcca taaagcagaa cggcctgctc
     1801 catgacaaat ctggctcccc aactaatgcc ccatgcagcc agcataacca gcataaagtg
     1861 cagtgtccgg tttgataggg ataagtccag ccttgcaaga agcggataca ggagtgcaaa
     1921 aaatggctat ctctagtaag gcctacccct taggctttat gcaacagaaa caataataat
     1981 ggagtcatga ccatgcctag gccgcggccg cgcgaattcg agctcggtac ccggggatcc
     2041 tctagagtcg acctgcaggc atgcaagctt gcggccgcgt cgtgactggg aaaaccctgg
     2101 cgactagtct tggactcctg ttgatagatc cagtaatgac ctcagaactc catctggatt
     2161 tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaatcca ggggtcccca
     2221 ataattacga tttacgtatt taaatttgac agcttatcat cgataaactg taatgcggta
     2281 gtttatcaca gttaaattgc taacgcagtc aggcaccgtg tatgaaatct aacaatgcgc
     2341 tcatcgtcat cctcggcacc gtcaccctgg atgctgtagg cataggcttg gttatgccgg
     2401 tactgccggg cctcttgcgg gatatcgtcc attccgacag catcgccagt cactatggcg
     2461 tgctgctagc gctatatgcg ttgatgcaat ttctatgcgc acccgttctc ggagcactgt
     2521 ccgaccgctt tggccgccgc ccagtcctgc tcgcttcgct acttggagcc actatcgact
     2581 acgcgatcat ggcgaccaca cccgtcctgt ggattctcta cgccggacgc atcgtggccg
     2641 gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg
     2701 aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag
     2761 gccccgtggc cgggggactg ttgggcgcca tctccctgca cgcaccattc cttgcggcgg
     2821 cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag tcgcataagg
     2881 gagagcgccg tccgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc
     2941 ggggcatgac cattgtggcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac
     3001 aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga
     3061 tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca
     3121 ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccg
     3181 acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc ttccccatta
     3241 tgattcttct cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc
     3301 aggtagatga cgaccatcag ggacagcttc aaggatcgct cgcggctctt accagcctaa
     3361 cttcgatcac tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg agcacatgga
     3421 acgggttggc atggattgta ggcgccgccc tataccttgt ctgcctcccc gcgttgcgtc
     3481 gcggtgcatg gagccgggcc acctcgacct gagacaattg tccttttccg ctgcataacc
     3541 ctgcttcggg gtcattatag cgattttttc ggtatatcca tcctttttcg cacgatatac
     3601 aggattttgc caaagggttc gtgtagactt tccttggtgt atccaacggc gtcagccggg
     3661 caggataggt gaagtaggcc cacccgcgag cgggtgttcc ttcttcactg tcccttattc
     3721 gcacctggcg gtgctcaacg ggaatcctgc tctgcgaggc tggccgtagg ccggccgcga
     3781 tgcaggtggc tgctgaaccc ccagccggaa ctgaccccac aaggccctag cgtttgcaat
     3841 gcaccaggtc atcattgacc caggcgtgtt ccaccaggcc gctgcctcgc aactcttcgc
     3901 aggcttcgcc gacctgctcg cgccacttct tcacgcgggt ggaatccgat ccgcacatga
     3961 ggcggaaggt ttccagcttg agcgggtacg gctcccggtg cgagctgaaa tagtcgaaca
     4021 tccgtcgggc cgtcggcgac agcttgcggt acttctccca tatgaatttc gtgtagtggt
     4081 cgccagcaaa cagcacgacg atttcctcgt cgatcaggac ctggcaacgg gacgttttct
     4141 tgccacggtc caggacgcgg aagcggtgca gcagcgacac cgattccagg tgcccaacgc
     4201 ggtcggacgt gaagcccatc gccgtcgcct gtaggcgcga caggcattcc tcggccttcg
     4261 tgtaataccg gccattgatc gaccagccca ggtcctggca aagctcgtag aacgtgaagg
     4321 tgatcggctc gccgataggg gtgcgcttcg cgtactccaa cacctgctgc cacaccagtt
     4381 cgtcatcgtc ggcccgcagc tcgacgccgg tgtaggtgat cttcacgtcc ttgttgacgt
     4441 ggaaaatgac cttgttttgc agcgcctcgc gcgggatttt cttgttgcgc gtggtgaaca
     4501 gggcagagcg ggccgtgtcg tttggcatcg ctcgcatcgt gtccggccac ggcgcaatat
     4561 cgaacaagga aagctgcatt tccttgatct gctgcttcgt gtgtttcagc aacgcggcct
     4621 gcttggcttc gctgacctgt tttgccaggt cctcgccggc ggtttttcgc ttcttggtcg
     4681 tcatagttcc tcgcgtgtcg atggtcatcg acttcgccaa acctgccgcc tcctgttcga
     4741 gacgacgcga acgctccacg gcggccgatg gcgcgggcag ggcaggggga gccagttgca
     4801 cgctgtcgcg ctcgatcttg gccgtagctt gctggactat cgagccgacg gactggaagg
     4861 tttcgcgggg cgcacgcatg acggtgcggc ttgcgatggt ttcggcatcc tcggcggaaa
     4921 accccgcgtc gatcagttct tgcctgtatg ccttccggtc aaacgtccga ttcattcacc
     4981 ctccttgcgg gattgccccg gaattaattc cccggatcga tccgtcgatc ttgatcccct
     5041 gcgccatcag atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac
     5101 cttaccagag ggcgccccag ctggcaattc cggttcgctt gctgtccata aaaccgccca
     5161 gtctagctat cgccatgtaa gcccactgca agctacctgc tttctctttg cgcttgcgtt
     5221 ttcccttgtc cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg
     5281 actggctttc tacgtggctg ccatttttgg ggtgaggccg ttcgcggccg aggggcgcag
     5341 cccctggggg gatgggaggc ccgcgttagc gggccgggag ggttcgagaa gggggggcac
     5401 cccccttcgg cgtgcgcggt cacgcgcaca gggcgcagcc ctggttaaaa acaaggttta
     5461 taaatattgg tttaaaagca ggttaaaaga caggttagcg gtggccgaaa aacgggcgga
     5521 aacccttgca aatgctggat tttctgcctg tggacagccc ctcaaatgtc aataggtgcg
     5581 cccctcatct gtcagcactc tgcccctcaa gtgtcaagga tcgcgcccct catctgtcag
     5641 tagtcgcgcc cctcaagtgt caataccgca gggcacttat ccccaggctt gtccacatca
     5701 tctgtgggaa actcgcgtaa aatcaggcgt tttcgccgat ttgcgaggct ggccagctcc
     5761 acgtcgccgg ccgaaatcga gcctgcccct catctgtcaa cgccgcgccg ggtgagtcgg
     5821 cccctcaagt gtcaacgtcc gcccctcatc tgtcagtgag ggccaagttt tccgcgaggt
     5881 atccacaacg ccggcggccc tacatggctc tgctgtagtg agtgggttgc gctccggcag
     5941 cggtcctgat cccccgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6001 cgcgcccagc tgtctagggc ggcggatttg tcctactcag gagagcgttc accgacaaac
     6061 aacagataaa acgaaaggcc cagtctttcg actgagcctt tcgttttatt tgatgcct
//
