LOCUS       40924_39423        5275 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA251, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5275)
  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  JOURNAL   Unpublished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 5275)
  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 5275)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5275)
  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            "Unpublished"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..5275
                     /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             381..1193
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            1363..1471
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(1485..2333)
                     /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(2323..3159)
                     /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(3673..4641)
                     /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(4665..4695)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     rep_origin      4751..5145
                     /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(5183..5269)
                     /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 gatttaaatt tgtgtctcaa aatctctgat
      301 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
      361 acagtaatac aaggggtgtt atgagccata ttcagcgtga aacgagctgt agccgtccgc
      421 gtctgaacag caacatggat gcggatctgt atggctataa atgggcgcgt gataacgtgg
      481 gtcagagcgg cgcgaccatt tatcgtctgt atggcaaacc ggatgcgccg gaactgtttc
      541 tgaaacatgg caaaggcagc gtggcgaacg atgtgaccga tgaaatggtg cgtctgaact
      601 ggctgaccga atttatgccg ctgccgacca ttaaacattt tattcgcacc ccggatgatg
      661 cgtggctgct gaccaccgcg attccgggca aaaccgcgtt tcaggtgctg gaagaatatc
      721 cggatagcgg cgaaaacatt gtggatgcgc tggccgtgtt tctgcgtcgt ctgcatagca
      781 ttccggtgtg caactgcccg tttaacagcg atcgtgtgtt tcgtctggcc caggcgcaga
      841 gccgtatgaa caacggcctg gtggatgcga gcgattttga tgatgaacgt aacggctggc
      901 cggtggaaca ggtgtggaaa gaaatgcata aactgctgcc gtttagcccg gatagcgtgg
      961 tgacccacgg cgattttagc ctggataacc tgattttcga tgaaggcaaa ctgattggct
     1021 gcattgatgt gggccgtgtg ggcattgcgg atcgttatca ggatctggcc attctgtgga
     1081 actgcctggg cgaatttagc ccgagcctgc aaaaacgtct gtttcagaaa tatggcattg
     1141 ataatccgga tatgaacaaa ctgcaatttc atctgatgct ggatgaattt ttctaataat
     1201 taattggacc gcggtccgcg cgttgtcctt ttccgctgca taaccctgct tcggggtcat
     1261 tatagcgatt ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag
     1321 ggttcgtgta gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt
     1381 aggcccaccc gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct
     1441 caacgggaat cctgctctgc gaggctggcc gtaggccggc ctcagcctgc cgccttgggc
     1501 cgggtgatgt cgtacttgcc cgccgcgaac tcggttaccg tccagcccag cgcgaccagc
     1561 tccggcaacg cctcgcgcac ccgctggcgg cgcttgcgca tggtcgaacc actggcctct
     1621 gacggccaga catagccgca caaggtatct atggaagcct tgccggtttt gccggggtcg
     1681 atccagccac acagccgctg gtgcagcagg cgggcggttt cgctgtccag cgcccgcacc
     1741 tcgtccatgc tgatgcgcac atgctggccg ccacccatga cggcctgcgc gatcaagggg
     1801 ttcagggcca cgtacaggcg cccgtccgcc tcgtcgctgg cgtactccga cagcagccga
     1861 aacccctgcc gcttgcggcc attctgggcg atgatggata ccttccaaag gcgctcgatg
     1921 cagtcctgta tgtgcttgag cgccccacca ctatcgacct ctgccccgat ttcctttgcc
     1981 agcgcccgat agctaccttt gaccacatgg cattcagcgg tgacggcctc ccacttgggt
     2041 tccaggaaca gccggagctg ccgtccgcct tcggtcttgg gttccgggcc aagcactagg
     2101 ccattaggcc cagccatggc caccagccct tgcaggatgc gcagatcatc agcgcccagc
     2161 ggctccgggc cgctgaactc gatccgcttg ccgtcgccgt agtcatacgt cacgtccagc
     2221 ttgctgcgct tgcgctcgcc ccgcttgagg gcacggaaca ggccgggggc cagacagtgc
     2281 gccgggtcgt gccggacgtg gctgaggctg tgcttgttct taggcttcac cacggggcac
     2341 ccccttgctc ttgcgctgcc tctccagcac ggcgggcttg agcaccccgc cgtcatgccg
     2401 cctgaaccac cgatcagcga acggtgcgcc atagttggcc ttgctcacac cgaagcggac
     2461 gaagaaccgg cgctggtcgt cgtccacacc ccattcctcg gcctcggcgc tggtcatgct
     2521 cgacaggtag gactgccagc ggatgttatc gaccagtacc gagctgcccc ggctggcctg
     2581 ctgctggtcg cctgcgccca tcatggccgc gcccttgctg gcatggtgca ggaacacgat
     2641 agagcacccg gtatcggcgg cgatggcctc catgcgaccg atgacctggg ccatggggcc
     2701 gctggcgttt tcttcctcga tgtggaaccg gcgcagcgtg tccagcacca tcaggcggcg
     2761 gccctcggcg gcgcgcttga ggccgtcgaa ccactccggg gccatgatgt tgggcaggct
     2821 gccgatcagc ggctggatca gcaggccgtc agccacggct tgccgttcct cggcgctgag
     2881 gtgcgcccca agggcgtgca ggcggtgatg aatggcggtg ggcgggtctt cggcgggcag
     2941 gtagatcacc gggccggtgg gcagttcgcc cacctccagc agatccggcc cgcctgcaat
     3001 ctgtgcggcc agttgcaggg ccagcatgga tttaccggca ccaccgggcg acaccagcgc
     3061 cccgaccgta ccggccacca tgttgggcaa aacgtagtcc agcggtggcg gcgctgctgc
     3121 gaacgcctcc agaatattga taggcttatg ggtagccatt gattgcctcc tttgcaggca
     3181 gttggtggtt aggcgctggc ggggtcacta cccccgccct gcgccgctct gagttcttcc
     3241 aggcactcgc gcagcgcctc gtattcgtcg tcggtcagcc agaacttgcg ctgacgcatc
     3301 cctttggcct tcatgcgctc ggcatatcgc gcttggcgta cagcgtcagg gctggccagc
     3361 aggtcgccgg tctgcttgtc cttttggtct ttcatatcag tcaccgagaa acttgccggg
     3421 gccgaaaggc ttgtcttcgc ggaacaagga caaggtgcag ccgtcaaggt taaggctggc
     3481 catatcagcg actgaaaagc ggccagcctc ggccttgttt gacgtataac caaagccacc
     3541 gggcaaccaa tagcccttgt cacttttgat caggtagacc gaccctgaag cgcttttttc
     3601 gtattccata aaaccccctt ctgtgcgtga gtactcatag tataacaggc gtgagtacca
     3661 acgcaagcac tacatgctga aatctggccc gcccctgtcc atgcctcgct ggcggggtgc
     3721 cggtgcccgt gccagctcgg cccgcgcaag ctggacgctg ggcagaccca tgaccttgct
     3781 gacggtgcgc tcgatgtaat ccgcttcgtg gccgggcttg cgctctgcca gcgctgggct
     3841 ggcctcggcc atggccttgc cgatttcctc ggcactgcgg ccccggctgg ccagcttctg
     3901 cgcggcgata aagtcgcact tgctgaggtc atcaccgaag cgcttgacca gcccggccat
     3961 ctcgctgcgg tactcgtcca gcgccgtgcg ccggtggcgg ctaagctgcc gctcgggcag
     4021 ttcgaggctg gccagcctgc gggccttctc ctgctgccgc tgggcctgct cgatctgctg
     4081 gccagcctgc tgcaccagcg ccgggccagc ggtggcggtc ttgcccttgg attcacgcag
     4141 cagcacccac ggctgataac cggcgcgggt ggtgtgcttg tccttgcggt tggtgaagcc
     4201 cgccaagcgg ccatagtggc ggctgtcggc gctggccggg tcggcgtcgt actcgctggc
     4261 cagcgtccgg gcaatctgcc cccgaagttc accgcctgcg gcgtcggcca ccttgaccca
     4321 tgcctgatag ttcttcgggc tggtttccac taccagggca ggctcccggc cctcggcttt
     4381 catgtcatcc aggtcaaact cgctgaggtc gtccaccagc accagaccat gccgctcctg
     4441 ctcggcgggc ctgatataca cgtcattgcc ctgggcattc atccgcttga gccatggcgt
     4501 gttctggagc acttcggcgg ctgaccattc ccggttcatc atctggccgg tggtggcgtc
     4561 cctgacgccg atatcgaagc gctcacagcc catggccttg agctgtcggc ctatggcctg
     4621 caaagtcctg tcgttcttca tcgggccacc aagcgattcc cacacattat acgagccgga
     4681 agcataaagt gtaaagccta gatccgaagg atgagccggg ctgaatgatc gaccgagaca
     4741 ggccctgcgg ggctgcacac gcgcccccac ccttcgggta gggggaaagg ccgctaaagc
     4801 ggctaaaagc gctccagcgt atttctgcgg ggtttggtgt ggggtttagc gggctttgcc
     4861 cgcctttccc cctgccgcgc agcggtgggg cggtgtgtag cctagcgcag cgaatagacc
     4921 agctatccgg cctctggccg ggcatattgg gcaagggcag cagcgcccca caagggcgct
     4981 gataaccgcg cctagtggat tattcttaga taatcatgga tggatttttc caacaccccg
     5041 ccagcccccg cccctgctgg gtttgcaggt ttgggggcgt gacagttatt gcaggggttc
     5101 gtgacagtta ttgcaggggg gcgtgacagt tattgcaggg gttcgtgaca gttagggcgc
     5161 gcccagctgt ctagggcggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
     5221 agataaaacg aaaggcccag tctttcgact gagcctttcg ttttatttga tgcct
//
