LOCUS       40924_39543        5334 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA451, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5334)
  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  JOURNAL   Unpublished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 5334)
  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 5334)
  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 (30-SEP-2014) Systems Biology Program, Centro Nacional de 
            Biotecnologia-CSIC, C/ Darwin 3, Campus de Cantoblanco, Madrid, 
            Madrid 28049, Spain
REFERENCE   4  (bases 1 to 5334)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5334)
  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            "Unpublished"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"; journalName: "Submitted 
            (30-SEP-2014) Systems Biology Program, Centro Nacional de 
            Biotecnologia-CSIC, C/ Darwin 3, Campus de Cantoblanco, Madrid, 
            Madrid 28049, Spain"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: ApE v. v2.0.40
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
            On Sep 30, 2014 this sequence version replaced JX560340.1.
FEATURES             Location/Qualifiers
     source          1..5334
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..32
                     /label=R24
                     /note="R24"
     misc_feature    56..112
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(121..144)
                     /label=F24
                     /note="F24"
     terminator      155..249
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             485..1273
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     oriT            1422..1530
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(1544..2392)
                     /codon_start=1
                     /label=RSF1010 RepC
                     /note="replication protein C of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /translation="VVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TAECHVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(2382..3218)
                     /codon_start=1
                     /label=RSF1010 RepA
                     /note="replication protein A of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             complement(3732..4700)
                     /codon_start=1
                     /label=RSF1010 RepB
                     /note="replication protein B of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     promoter        complement(4724..4754)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     rep_origin      4810..5204
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     terminator      complement(5242..5328)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ttaattaaag cggataacaa tttcacacag gaggccgcct aggccgcggc cgcgcgaatt
       61 cgagctcggt acccggggat cctctagagt cgacctgcag gcatgcaagc ttgcggccgc
      121 gtcgtgactg ggaaaaccct ggcgactagt cttggactcc tgttgataga tccagtaatg
      181 acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt
      241 ggtgagaatc caggggtccc caataattac gatttacgta tttaaatgaa ccttgaccga
      301 acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg
      361 ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt
      421 gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa
      481 catcatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt
      541 catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga
      601 tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga
      661 tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga
      721 gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg
      781 gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc
      841 aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag
      901 agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga
      961 acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg
     1021 ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac
     1081 cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca
     1141 gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt
     1201 ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa
     1261 ggtagtcggc aaataagaca attgtccttt tccgctgcat aaccctgctt cggggtcatt
     1321 atagcgattt tttcggtata tccatccttt ttcgcacgat atacaggatt ttgccaaagg
     1381 gttcgtgtag actttccttg gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta
     1441 ggcccacccg cgagcgggtg ttccttcttc actgtccctt attcgcacct ggcggtgctc
     1501 aacgggaatc ctgctctgcg aggctggccg taggccggcc tcagcctgcc gccttgggcc
     1561 gggtgatgtc gtacttgccc gccgcgaact cggttaccgt ccagcccagc gcgaccagct
     1621 ccggcaacgc ctcgcgcacc cgctggcggc gcttgcgcat ggtcgaacca ctggcctctg
     1681 acggccagac atagccgcac aaggtatcta tggaagcctt gccggttttg ccggggtcga
     1741 tccagccaca cagccgctgg tgcagcaggc gggcggtttc gctgtccagc gcccgcacct
     1801 cgtccatgct gatgcgcaca tgctggccgc cacccatgac ggcctgcgcg atcaaggggt
     1861 tcagggccac gtacaggcgc ccgtccgcct cgtcgctggc gtactccgac agcagccgaa
     1921 acccctgccg cttgcggcca ttctgggcga tgatggatac cttccaaagg cgctcgatgc
     1981 agtcctgtat gtgcttgagc gccccaccac tatcgacctc tgccccgatt tcctttgcca
     2041 gcgcccgata gctacctttg accacatggc attcagcggt gacggcctcc cacttgggtt
     2101 ccaggaacag ccggagctgc cgtccgcctt cggtcttggg ttccgggcca agcactaggc
     2161 cattaggccc agccatggcc accagccctt gcaggatgcg cagatcatca gcgcccagcg
     2221 gctccgggcc gctgaactcg atccgcttgc cgtcgccgta gtcatacgtc acgtccagct
     2281 tgctgcgctt gcgctcgccc cgcttgaggg cacggaacag gccgggggcc agacagtgcg
     2341 ccgggtcgtg ccggacgtgg ctgaggctgt gcttgttctt aggcttcacc acggggcacc
     2401 cccttgctct tgcgctgcct ctccagcacg gcgggcttga gcaccccgcc gtcatgccgc
     2461 ctgaaccacc gatcagcgaa cggtgcgcca tagttggcct tgctcacacc gaagcggacg
     2521 aagaaccggc gctggtcgtc gtccacaccc cattcctcgg cctcggcgct ggtcatgctc
     2581 gacaggtagg actgccagcg gatgttatcg accagtaccg agctgccccg gctggcctgc
     2641 tgctggtcgc ctgcgcccat catggccgcg cccttgctgg catggtgcag gaacacgata
     2701 gagcacccgg tatcggcggc gatggcctcc atgcgaccga tgacctgggc catggggccg
     2761 ctggcgtttt cttcctcgat gtggaaccgg cgcagcgtgt ccagcaccat caggcggcgg
     2821 ccctcggcgg cgcgcttgag gccgtcgaac cactccgggg ccatgatgtt gggcaggctg
     2881 ccgatcagcg gctggatcag caggccgtca gccacggctt gccgttcctc ggcgctgagg
     2941 tgcgccccaa gggcgtgcag gcggtgatga atggcggtgg gcgggtcttc ggcgggcagg
     3001 tagatcaccg ggccggtggg cagttcgccc acctccagca gatccggccc gcctgcaatc
     3061 tgtgcggcca gttgcagggc cagcatggat ttaccggcac caccgggcga caccagcgcc
     3121 ccgaccgtac cggccaccat gttgggcaaa acgtagtcca gcggtggcgg cgctgctgcg
     3181 aacgcctcca gaatattgat aggcttatgg gtagccattg attgcctcct ttgcaggcag
     3241 ttggtggtta ggcgctggcg gggtcactac ccccgccctg cgccgctctg agttcttcca
     3301 ggcactcgcg cagcgcctcg tattcgtcgt cggtcagcca gaacttgcgc tgacgcatcc
     3361 ctttggcctt catgcgctcg gcatatcgcg cttggcgtac agcgtcaggg ctggccagca
     3421 ggtcgccggt ctgcttgtcc ttttggtctt tcatatcagt caccgagaaa cttgccgggg
     3481 ccgaaaggct tgtcttcgcg gaacaaggac aaggtgcagc cgtcaaggtt aaggctggcc
     3541 atatcagcga ctgaaaagcg gccagcctcg gccttgtttg acgtataacc aaagccaccg
     3601 ggcaaccaat agcccttgtc acttttgatc aggtagaccg accctgaagc gcttttttcg
     3661 tattccataa aacccccttc tgtgcgtgag tactcatagt ataacaggcg tgagtaccaa
     3721 cgcaagcact acatgctgaa atctggcccg cccctgtcca tgcctcgctg gcggggtgcc
     3781 ggtgcccgtg ccagctcggc ccgcgcaagc tggacgctgg gcagacccat gaccttgctg
     3841 acggtgcgct cgatgtaatc cgcttcgtgg ccgggcttgc gctctgccag cgctgggctg
     3901 gcctcggcca tggccttgcc gatttcctcg gcactgcggc cccggctggc cagcttctgc
     3961 gcggcgataa agtcgcactt gctgaggtca tcaccgaagc gcttgaccag cccggccatc
     4021 tcgctgcggt actcgtccag cgccgtgcgc cggtggcggc taagctgccg ctcgggcagt
     4081 tcgaggctgg ccagcctgcg ggccttctcc tgctgccgct gggcctgctc gatctgctgg
     4141 ccagcctgct gcaccagcgc cgggccagcg gtggcggtct tgcccttgga ttcacgcagc
     4201 agcacccacg gctgataacc ggcgcgggtg gtgtgcttgt ccttgcggtt ggtgaagccc
     4261 gccaagcggc catagtggcg gctgtcggcg ctggccgggt cggcgtcgta ctcgctggcc
     4321 agcgtccggg caatctgccc ccgaagttca ccgcctgcgg cgtcggccac cttgacccat
     4381 gcctgatagt tcttcgggct ggtttccact accagggcag gctcccggcc ctcggctttc
     4441 atgtcatcca ggtcaaactc gctgaggtcg tccaccagca ccagaccatg ccgctcctgc
     4501 tcggcgggcc tgatatacac gtcattgccc tgggcattca tccgcttgag ccatggcgtg
     4561 ttctggagca cttcggcggc tgaccattcc cggttcatca tctggccggt ggtggcgtcc
     4621 ctgacgccga tatcgaagcg ctcacagccc atggccttga gctgtcggcc tatggcctgc
     4681 aaagtcctgt cgttcttcat cgggccacca agcgattccc acacattata cgagccggaa
     4741 gcataaagtg taaagcctag atccgaagga tgagccgggc tgaatgatcg accgagacag
     4801 gccctgcggg gctgcacacg cgcccccacc cttcgggtag ggggaaaggc cgctaaagcg
     4861 gctaaaagcg ctccagcgta tttctgcggg gtttggtgtg gggtttagcg ggctttgccc
     4921 gcctttcccc ctgccgcgca gcggtggggc ggtgtgtagc ctagcgcagc gaatagacca
     4981 gctatccggc ctctggccgg gcatattggg caagggcagc agcgccccac aagggcgctg
     5041 ataaccgcgc ctagtggatt attcttagat aatcatggat ggatttttcc aacaccccgc
     5101 cagcccccgc ccctgctggg tttgcaggtt tgggggcgtg acagttattg caggggttcg
     5161 tgacagttat tgcagggggg cgtgacagtt attgcagggg ttcgtgacag ttagggcgcg
     5221 cccagctgtc tagggcggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     5281 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcct
//
