LOCUS       40924_39288        4525 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA227C, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4525)
  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  JOURNAL   Unpublished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 4525)
  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 4525)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4525)
  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            "Unpublished"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..4525
                     /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"
     CDS             127..843
                     /codon_start=1
                     /label=ECFP
                     /note="enhanced CFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      857..951
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1083..1895
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            2065..2173
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(2237..3382)
                     /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(3775..4306)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(4433..4519)
                     /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 ttaggaggaa
      121 aaacatatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      181 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      241 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
      301 cccaccctcg tgaccaccct gacctggggc gtgcagtgct tcagccgcta ccccgaccac
      361 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
      421 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
      481 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
      541 gggcacaagc tggagtacaa ctacatcagc cacaacgtct atatcaccgc cgacaagcag
      601 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
      661 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
      721 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
      781 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
      841 aagtaaacta gtcttggact cctgttgata gatccagtaa tgacctcaga actccatctg
      901 gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaggggtc
      961 cccaataatt acgatttaaa tttgtgtctc aaaatctctg atgttacatt gcacaagata
     1021 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     1081 ttatgagcca tattcagcgt gaaacgagct gtagccgtcc gcgtctgaac agcaacatgg
     1141 atgcggatct gtatggctat aaatgggcgc gtgataacgt gggtcagagc ggcgcgacca
     1201 tttatcgtct gtatggcaaa ccggatgcgc cggaactgtt tctgaaacat ggcaaaggca
     1261 gcgtggcgaa cgatgtgacc gatgaaatgg tgcgtctgaa ctggctgacc gaatttatgc
     1321 cgctgccgac cattaaacat tttattcgca ccccggatga tgcgtggctg ctgaccaccg
     1381 cgattccggg caaaaccgcg tttcaggtgc tggaagaata tccggatagc ggcgaaaaca
     1441 ttgtggatgc gctggccgtg tttctgcgtc gtctgcatag cattccggtg tgcaactgcc
     1501 cgtttaacag cgatcgtgtg tttcgtctgg cccaggcgca gagccgtatg aacaacggcc
     1561 tggtggatgc gagcgatttt gatgatgaac gtaacggctg gccggtggaa caggtgtgga
     1621 aagaaatgca taaactgctg ccgtttagcc cggatagcgt ggtgacccac ggcgatttta
     1681 gcctggataa cctgattttc gatgaaggca aactgattgg ctgcattgat gtgggccgtg
     1741 tgggcattgc ggatcgttat caggatctgg ccattctgtg gaactgcctg ggcgaattta
     1801 gcccgagcct gcaaaaacgt ctgtttcaga aatatggcat tgataatccg gatatgaaca
     1861 aactgcaatt tcatctgatg ctggatgaat ttttctaata attaattgga ccgcggtccg
     1921 cgcgttgtcc ttttccgctg cataaccctg cttcggggtc attatagcga ttttttcggt
     1981 atatccatcc tttttcgcac gatatacagg attttgccaa agggttcgtg tagactttcc
     2041 ttggtgtatc caacggcgtc agccgggcag gataggtgaa gtaggcccac ccgcgagcgg
     2101 gtgttccttc ttcactgtcc cttattcgca cctggcggtg ctcaacggga atcctgctct
     2161 gcgaggctgg ccgtaggccg gccgcgatgc aggtggctgc tgaaccccca gccggaactg
     2221 accccacaag gccctagcgt ttgcaatgca ccaggtcatc attgacccag gcgtgttcca
     2281 ccaggccgct gcctcgcaac tcttcgcagg cttcgccgac ctgctcgcgc cacttcttca
     2341 cgcgggtgga atccgatccg cacatgaggc ggaaggtttc cagcttgagc gggtacggct
     2401 cccggtgcga gctgaaatag tcgaacatcc gtcgggccgt cggcgacagc ttgcggtact
     2461 tctcccatat gaatttcgtg tagtggtcgc cagcaaacag cacgacgatt tcctcgtcga
     2521 tcaggacctg gcaacgggac gttttcttgc cacggtccag gacgcggaag cggtgcagca
     2581 gcgacaccga ttccaggtgc ccaacgcggt cggacgtgaa gcccatcgcc gtcgcctgta
     2641 ggcgcgacag gcattcctcg gccttcgtgt aataccggcc attgatcgac cagcccaggt
     2701 cctggcaaag ctcgtagaac gtgaaggtga tcggctcgcc gataggggtg cgcttcgcgt
     2761 actccaacac ctgctgccac accagttcgt catcgtcggc ccgcagctcg acgccggtgt
     2821 aggtgatctt cacgtccttg ttgacgtgga aaatgacctt gttttgcagc gcctcgcgcg
     2881 ggattttctt gttgcgcgtg gtgaacaggg cagagcgggc cgtgtcgttt ggcatcgctc
     2941 gcatcgtgtc cggccacggc gcaatatcga acaaggaaag ctgcatttcc ttgatctgct
     3001 gcttcgtgtg tttcagcaac gcggcctgct tggcttcgct gacctgtttt gccaggtcct
     3061 cgccggcggt ttttcgcttc ttggtcgtca tagttcctcg cgtgtcgatg gtcatcgact
     3121 tcgccaaacc tgccgcctcc tgttcgagac gacgcgaacg ctccacggcg gccgatggcg
     3181 cgggcagggc agggggagcc agttgcacgc tgtcgcgctc gatcttggcc gtagcttgct
     3241 ggactatcga gccgacggac tggaaggttt cgcggggcgc acgcatgacg gtgcggcttg
     3301 cgatggtttc ggcatcctcg gcggaaaacc ccgcgtcgat cagttcttgc ctgtatgcct
     3361 tccggtcaaa cgtccgattc attcaccctc cttgcgggat tgccccggaa ttaattcccc
     3421 ggatcgatcc gtcgatcttg atcccctgcg ccatcagatc cttggcggca agaaagccat
     3481 ccagtttact ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg
     3541 ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc
     3601 tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt
     3661 catccggggt cagcaccgtt tctgcggact ggctttctac gtggctgcca tttttggggt
     3721 gaggccgttc gcggccgagg ggcgcagccc ctggggggat gggaggcccg cgttagcggg
     3781 ccgggagggt tcgagaaggg ggggcacccc ccttcggcgt gcgcggtcac gcgcacaggg
     3841 cgcagccctg gttaaaaaca aggtttataa atattggttt aaaagcaggt taaaagacag
     3901 gttagcggtg gccgaaaaac gggcggaaac ccttgcaaat gctggatttt ctgcctgtgg
     3961 acagcccctc aaatgtcaat aggtgcgccc ctcatctgtc agcactctgc ccctcaagtg
     4021 tcaaggatcg cgcccctcat ctgtcagtag tcgcgcccct caagtgtcaa taccgcaggg
     4081 cacttatccc caggcttgtc cacatcatct gtgggaaact cgcgtaaaat caggcgtttt
     4141 cgccgatttg cgaggctggc cagctccacg tcgccggccg aaatcgagcc tgcccctcat
     4201 ctgtcaacgc cgcgccgggt gagtcggccc ctcaagtgtc aacgtccgcc cctcatctgt
     4261 cagtgagggc caagttttcc gcgaggtatc cacaacgccg gcggccctac atggctctgc
     4321 tgtagtgagt gggttgcgct ccggcagcgg tcctgatccc ccgcagaaaa aaaggatctc
     4381 aagaagatcc tttgatcttt tctacggcgc gcccagctgt ctagggcggc ggatttgtcc
     4441 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact
     4501 gagcctttcg ttttatttga tgcct
//
