LOCUS       40924_39443        5626 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA328, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5626)
  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  JOURNAL   Unpublished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 5626)
  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 5626)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5626)
  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            "Unpublished"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..5626
                     /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=F24
                     /note="F24"
     terminator      2113..2207
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        2255..2357
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             2358..3014
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     oriT            3166..3274
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(3338..4483)
                     /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(4876..5407)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(5534..5620)
                     /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 tttaaattgg cgaaaatgag acgttgatcg gcacgtaaga ggttccaact
     2281 ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag
     2341 gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc
     2401 aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc
     2461 agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt
     2521 tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg gaatttcgta
     2581 tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt
     2641 tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc
     2701 agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc
     2761 ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca
     2821 gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca
     2881 aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg
     2941 tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt
     3001 ggcagggcgg ggcgtaattt gacttttgtc cttttccgct gcataaccct gcttcggggt
     3061 cattatagcg attttttcgg tatatccatc ctttttcgca cgatatacag gattttgcca
     3121 aagggttcgt gtagactttc cttggtgtat ccaacggcgt cagccgggca ggataggtga
     3181 agtaggccca cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc acctggcggt
     3241 gctcaacggg aatcctgctc tgcgaggctg gccgtaggcc ggccgcgatg caggtggctg
     3301 ctgaaccccc agccggaact gaccccacaa ggccctagcg tttgcaatgc accaggtcat
     3361 cattgaccca ggcgtgttcc accaggccgc tgcctcgcaa ctcttcgcag gcttcgccga
     3421 cctgctcgcg ccacttcttc acgcgggtgg aatccgatcc gcacatgagg cggaaggttt
     3481 ccagcttgag cgggtacggc tcccggtgcg agctgaaata gtcgaacatc cgtcgggccg
     3541 tcggcgacag cttgcggtac ttctcccata tgaatttcgt gtagtggtcg ccagcaaaca
     3601 gcacgacgat ttcctcgtcg atcaggacct ggcaacggga cgttttcttg ccacggtcca
     3661 ggacgcggaa gcggtgcagc agcgacaccg attccaggtg cccaacgcgg tcggacgtga
     3721 agcccatcgc cgtcgcctgt aggcgcgaca ggcattcctc ggccttcgtg taataccggc
     3781 cattgatcga ccagcccagg tcctggcaaa gctcgtagaa cgtgaaggtg atcggctcgc
     3841 cgataggggt gcgcttcgcg tactccaaca cctgctgcca caccagttcg tcatcgtcgg
     3901 cccgcagctc gacgccggtg taggtgatct tcacgtcctt gttgacgtgg aaaatgacct
     3961 tgttttgcag cgcctcgcgc gggattttct tgttgcgcgt ggtgaacagg gcagagcggg
     4021 ccgtgtcgtt tggcatcgct cgcatcgtgt ccggccacgg cgcaatatcg aacaaggaaa
     4081 gctgcatttc cttgatctgc tgcttcgtgt gtttcagcaa cgcggcctgc ttggcttcgc
     4141 tgacctgttt tgccaggtcc tcgccggcgg tttttcgctt cttggtcgtc atagttcctc
     4201 gcgtgtcgat ggtcatcgac ttcgccaaac ctgccgcctc ctgttcgaga cgacgcgaac
     4261 gctccacggc ggccgatggc gcgggcaggg cagggggagc cagttgcacg ctgtcgcgct
     4321 cgatcttggc cgtagcttgc tggactatcg agccgacgga ctggaaggtt tcgcggggcg
     4381 cacgcatgac ggtgcggctt gcgatggttt cggcatcctc ggcggaaaac cccgcgtcga
     4441 tcagttcttg cctgtatgcc ttccggtcaa acgtccgatt cattcaccct ccttgcggga
     4501 ttgccccgga attaattccc cggatcgatc cgtcgatctt gatcccctgc gccatcagat
     4561 ccttggcggc aagaaagcca tccagtttac tttgcagggc ttcccaacct taccagaggg
     4621 cgccccagct ggcaattccg gttcgcttgc tgtccataaa accgcccagt ctagctatcg
     4681 ccatgtaagc ccactgcaag ctacctgctt tctctttgcg cttgcgtttt cccttgtcca
     4741 gatagcccag tagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta
     4801 cgtggctgcc atttttgggg tgaggccgtt cgcggccgag gggcgcagcc cctgggggga
     4861 tgggaggccc gcgttagcgg gccgggaggg ttcgagaagg gggggcaccc cccttcggcg
     4921 tgcgcggtca cgcgcacagg gcgcagccct ggttaaaaac aaggtttata aatattggtt
     4981 taaaagcagg ttaaaagaca ggttagcggt ggccgaaaaa cgggcggaaa cccttgcaaa
     5041 tgctggattt tctgcctgtg gacagcccct caaatgtcaa taggtgcgcc cctcatctgt
     5101 cagcactctg cccctcaagt gtcaaggatc gcgcccctca tctgtcagta gtcgcgcccc
     5161 tcaagtgtca ataccgcagg gcacttatcc ccaggcttgt ccacatcatc tgtgggaaac
     5221 tcgcgtaaaa tcaggcgttt tcgccgattt gcgaggctgg ccagctccac gtcgccggcc
     5281 gaaatcgagc ctgcccctca tctgtcaacg ccgcgccggg tgagtcggcc cctcaagtgt
     5341 caacgtccgc ccctcatctg tcagtgaggg ccaagttttc cgcgaggtat ccacaacgcc
     5401 ggcggcccta catggctctg ctgtagtgag tgggttgcgc tccggcagcg gtcctgatcc
     5461 cccgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacggcg cgcccagctg
     5521 tctagggcgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     5581 gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcct
//
