LOCUS       40924_39488        3882 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA421, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3882)
  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  JOURNAL   Unpublished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 3882)
  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 3882)
  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 3882)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3882)
  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            "Unpublished"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 JX560337.1.
FEATURES             Location/Qualifiers
     source          1..3882
                     /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(1594..2739)
                     /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(3132..3663)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(3790..3876)
                     /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 gcgatgcagg tggctgctga
     1561 acccccagcc ggaactgacc ccacaaggcc ctagcgtttg caatgcacca ggtcatcatt
     1621 gacccaggcg tgttccacca ggccgctgcc tcgcaactct tcgcaggctt cgccgacctg
     1681 ctcgcgccac ttcttcacgc gggtggaatc cgatccgcac atgaggcgga aggtttccag
     1741 cttgagcggg tacggctccc ggtgcgagct gaaatagtcg aacatccgtc gggccgtcgg
     1801 cgacagcttg cggtacttct cccatatgaa tttcgtgtag tggtcgccag caaacagcac
     1861 gacgatttcc tcgtcgatca ggacctggca acgggacgtt ttcttgccac ggtccaggac
     1921 gcggaagcgg tgcagcagcg acaccgattc caggtgccca acgcggtcgg acgtgaagcc
     1981 catcgccgtc gcctgtaggc gcgacaggca ttcctcggcc ttcgtgtaat accggccatt
     2041 gatcgaccag cccaggtcct ggcaaagctc gtagaacgtg aaggtgatcg gctcgccgat
     2101 aggggtgcgc ttcgcgtact ccaacacctg ctgccacacc agttcgtcat cgtcggcccg
     2161 cagctcgacg ccggtgtagg tgatcttcac gtccttgttg acgtggaaaa tgaccttgtt
     2221 ttgcagcgcc tcgcgcggga ttttcttgtt gcgcgtggtg aacagggcag agcgggccgt
     2281 gtcgtttggc atcgctcgca tcgtgtccgg ccacggcgca atatcgaaca aggaaagctg
     2341 catttccttg atctgctgct tcgtgtgttt cagcaacgcg gcctgcttgg cttcgctgac
     2401 ctgttttgcc aggtcctcgc cggcggtttt tcgcttcttg gtcgtcatag ttcctcgcgt
     2461 gtcgatggtc atcgacttcg ccaaacctgc cgcctcctgt tcgagacgac gcgaacgctc
     2521 cacggcggcc gatggcgcgg gcagggcagg gggagccagt tgcacgctgt cgcgctcgat
     2581 cttggccgta gcttgctgga ctatcgagcc gacggactgg aaggtttcgc ggggcgcacg
     2641 catgacggtg cggcttgcga tggtttcggc atcctcggcg gaaaaccccg cgtcgatcag
     2701 ttcttgcctg tatgccttcc ggtcaaacgt ccgattcatt caccctcctt gcgggattgc
     2761 cccggaatta attccccgga tcgatccgtc gatcttgatc ccctgcgcca tcagatcctt
     2821 ggcggcaaga aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc
     2881 ccagctggca attccggttc gcttgctgtc cataaaaccg cccagtctag ctatcgccat
     2941 gtaagcccac tgcaagctac ctgctttctc tttgcgcttg cgttttccct tgtccagata
     3001 gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc tttctacgtg
     3061 gctgccattt ttggggtgag gccgttcgcg gccgaggggc gcagcccctg gggggatggg
     3121 aggcccgcgt tagcgggccg ggagggttcg agaagggggg gcacccccct tcggcgtgcg
     3181 cggtcacgcg cacagggcgc agccctggtt aaaaacaagg tttataaata ttggtttaaa
     3241 agcaggttaa aagacaggtt agcggtggcc gaaaaacggg cggaaaccct tgcaaatgct
     3301 ggattttctg cctgtggaca gcccctcaaa tgtcaatagg tgcgcccctc atctgtcagc
     3361 actctgcccc tcaagtgtca aggatcgcgc ccctcatctg tcagtagtcg cgcccctcaa
     3421 gtgtcaatac cgcagggcac ttatccccag gcttgtccac atcatctgtg ggaaactcgc
     3481 gtaaaatcag gcgttttcgc cgatttgcga ggctggccag ctccacgtcg ccggccgaaa
     3541 tcgagcctgc ccctcatctg tcaacgccgc gccgggtgag tcggcccctc aagtgtcaac
     3601 gtccgcccct catctgtcag tgagggccaa gttttccgcg aggtatccac aacgccggcg
     3661 gccctacatg gctctgctgt agtgagtggg ttgcgctccg gcagcggtcc tgatcccccg
     3721 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggcgcgcc cagctgtcta
     3781 gggcggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa
     3841 ggcccagtct ttcgactgag cctttcgttt tatttgatgc ct
//
