LOCUS       40924_39553        4160 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA521, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4160)
  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  JOURNAL   Unpublished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 4160)
  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 4160)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4160)
  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            "Unpublished"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..4160
                     /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             364..1551
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     oriT            1700..1808
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(1872..3017)
                     /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(3410..3941)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(4068..4154)
                     /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 tttaaatttg acagcttatc
      301 atcgataaac tgtaatgcgg tagtttatca cagttaaatt gctaacgcag tcaggcaccg
      361 tgtatgaaat ctaacaatgc gctcatcgtc atcctcggca ccgtcaccct ggatgctgta
      421 ggcataggct tggttatgcc ggtactgccg ggcctcttgc gggatatcgt ccattccgac
      481 agcatcgcca gtcactatgg cgtgctgcta gcgctatatg cgttgatgca atttctatgc
      541 gcacccgttc tcggagcact gtccgaccgc tttggccgcc gcccagtcct gctcgcttcg
      601 ctacttggag ccactatcga ctacgcgatc atggcgacca cacccgtcct gtggattctc
      661 tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat
      721 atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt
      781 ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccctg
      841 cacgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc
      901 ctaatgcagg agtcgcataa gggagagcgc cgtccgatgc ccttgagagc cttcaaccca
      961 gtcagctcct tccggtgggc gcggggcatg accattgtgg ccgcacttat gactgtcttc
     1021 tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac
     1081 cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac
     1141 gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc
     1201 attatcgccg gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga
     1261 ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg
     1321 ttgcaggcca tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggatcg
     1381 ctcgcggctc ttaccagcct aacttcgatc actggaccgc tgatcgtcac ggcgatttat
     1441 gccgcctcgg cgagcacatg gaacgggttg gcatggattg taggcgccgc cctatacctt
     1501 gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac ctgagacaat
     1561 tgtccttttc cgctgcataa ccctgcttcg gggtcattat agcgattttt tcggtatatc
     1621 catccttttt cgcacgatat acaggatttt gccaaagggt tcgtgtagac tttccttggt
     1681 gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt
     1741 ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag
     1801 gctggccgta ggccggccgc gatgcaggtg gctgctgaac ccccagccgg aactgacccc
     1861 acaaggccct agcgtttgca atgcaccagg tcatcattga cccaggcgtg ttccaccagg
     1921 ccgctgcctc gcaactcttc gcaggcttcg ccgacctgct cgcgccactt cttcacgcgg
     1981 gtggaatccg atccgcacat gaggcggaag gtttccagct tgagcgggta cggctcccgg
     2041 tgcgagctga aatagtcgaa catccgtcgg gccgtcggcg acagcttgcg gtacttctcc
     2101 catatgaatt tcgtgtagtg gtcgccagca aacagcacga cgatttcctc gtcgatcagg
     2161 acctggcaac gggacgtttt cttgccacgg tccaggacgc ggaagcggtg cagcagcgac
     2221 accgattcca ggtgcccaac gcggtcggac gtgaagccca tcgccgtcgc ctgtaggcgc
     2281 gacaggcatt cctcggcctt cgtgtaatac cggccattga tcgaccagcc caggtcctgg
     2341 caaagctcgt agaacgtgaa ggtgatcggc tcgccgatag gggtgcgctt cgcgtactcc
     2401 aacacctgct gccacaccag ttcgtcatcg tcggcccgca gctcgacgcc ggtgtaggtg
     2461 atcttcacgt ccttgttgac gtggaaaatg accttgtttt gcagcgcctc gcgcgggatt
     2521 ttcttgttgc gcgtggtgaa cagggcagag cgggccgtgt cgtttggcat cgctcgcatc
     2581 gtgtccggcc acggcgcaat atcgaacaag gaaagctgca tttccttgat ctgctgcttc
     2641 gtgtgtttca gcaacgcggc ctgcttggct tcgctgacct gttttgccag gtcctcgccg
     2701 gcggtttttc gcttcttggt cgtcatagtt cctcgcgtgt cgatggtcat cgacttcgcc
     2761 aaacctgccg cctcctgttc gagacgacgc gaacgctcca cggcggccga tggcgcgggc
     2821 agggcagggg gagccagttg cacgctgtcg cgctcgatct tggccgtagc ttgctggact
     2881 atcgagccga cggactggaa ggtttcgcgg ggcgcacgca tgacggtgcg gcttgcgatg
     2941 gtttcggcat cctcggcgga aaaccccgcg tcgatcagtt cttgcctgta tgccttccgg
     3001 tcaaacgtcc gattcattca ccctccttgc gggattgccc cggaattaat tccccggatc
     3061 gatccgtcga tcttgatccc ctgcgccatc agatccttgg cggcaagaaa gccatccagt
     3121 ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat tccggttcgc
     3181 ttgctgtcca taaaaccgcc cagtctagct atcgccatgt aagcccactg caagctacct
     3241 gctttctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg acattcatcc
     3301 ggggtcagca ccgtttctgc ggactggctt tctacgtggc tgccattttt ggggtgaggc
     3361 cgttcgcggc cgaggggcgc agcccctggg gggatgggag gcccgcgtta gcgggccggg
     3421 agggttcgag aagggggggc accccccttc ggcgtgcgcg gtcacgcgca cagggcgcag
     3481 ccctggttaa aaacaaggtt tataaatatt ggtttaaaag caggttaaaa gacaggttag
     3541 cggtggccga aaaacgggcg gaaacccttg caaatgctgg attttctgcc tgtggacagc
     3601 ccctcaaatg tcaataggtg cgcccctcat ctgtcagcac tctgcccctc aagtgtcaag
     3661 gatcgcgccc ctcatctgtc agtagtcgcg cccctcaagt gtcaataccg cagggcactt
     3721 atccccaggc ttgtccacat catctgtggg aaactcgcgt aaaatcaggc gttttcgccg
     3781 atttgcgagg ctggccagct ccacgtcgcc ggccgaaatc gagcctgccc ctcatctgtc
     3841 aacgccgcgc cgggtgagtc ggcccctcaa gtgtcaacgt ccgcccctca tctgtcagtg
     3901 agggccaagt tttccgcgag gtatccacaa cgccggcggc cctacatggc tctgctgtag
     3961 tgagtgggtt gcgctccggc agcggtcctg atcccccgca gaaaaaaagg atctcaagaa
     4021 gatcctttga tcttttctac ggcgcgccca gctgtctagg gcggcggatt tgtcctactc
     4081 aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttt cgactgagcc
     4141 tttcgtttta tttgatgcct
//
