LOCUS       40924_39508        3631 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA432, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3631)
  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  JOURNAL   Unpublished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 3631)
  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 3631)
  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 3631)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3631)
  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            "Unpublished"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 JX560363.1.
FEATURES             Location/Qualifiers
     source          1..3631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    125..146
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        161..191
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    199..215
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     223..239
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(252..308)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(309..325)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(329..352)
                     /label=F24
                     /note="F24"
     terminator      604..698
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             934..1722
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     oriT            1871..1979
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(1993..2652)
                     /codon_start=1
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      complement(2653..3424)
                     /direction=LEFT
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      complement(3539..3625)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ttaattaaga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
       61 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
      121 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
      181 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
      241 atgattacgc caagcttgca tgcctgcagg tcgactctag aggatccccg ggtaccgagc
      301 tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
      361 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
      421 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
      481 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
      541 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgaactagtc
      601 ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa
      661 cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aggggtcccc aataattacg
      721 atttacgtat ttaaatgaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt
      781 ttcatggctt gttatgactg tttttttggg gtacagtcta tgcctcgggc atccaagcag
      841 caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag
      901 cagggcagtc gccctaaaac aaagttaaac atcatgaggg aagcggtgat cgccgaagta
      961 tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg
     1021 gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat
     1081 ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac
     1141 cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc
     1201 attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt
     1261 ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt
     1321 gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg
     1381 gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc
     1441 ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg
     1501 ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc
     1561 gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag
     1621 gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt
     1681 gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataagacaa ttgtcctttt
     1741 ccgctgcata accctgcttc ggggtcatta tagcgatttt ttcggtatat ccatcctttt
     1801 tcgcacgata tacaggattt tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac
     1861 ggcgtcagcc gggcaggata ggtgaagtag gcccacccgc gagcgggtgt tccttcttca
     1921 ctgtccctta ttcgcacctg gcggtgctca acgggaatcc tgctctgcga ggctggccgt
     1981 aggccggccc taccggcgcg gcagcgttac ccgtgtcggc ggctccaacg gctcgccatc
     2041 gtccagaaaa cacggctcat cgggcatcgg caggcgctgc tgcccgcgcc gttcccattc
     2101 ctccgtttcg gtcaaggctg gcaggtctgg ttccatgccc ggaatgccgg gctggctggg
     2161 cggctcctcg ccggggccgg tcggtagttg ctgctcgccc ggatacaggg tcgggatgcg
     2221 gcgcaggtcg ccatgcccca acagcgattc gtcctggtcg tcgtgatcaa ccaccacggc
     2281 ggcactgaac accgacaggc gcaactggtc gcggggctgg ccccacgcca cgcggtcatt
     2341 gaccacgtag gccgacacgg tgccggggcc gttgagcttc acgacggaga tccagcgctc
     2401 ggccaccaag tccttgactg cgtattggac cgtccgcaaa gaacgtccga tgagcttgga
     2461 aagtgtcttc tggctgacca ccacggcgtt ctggtggccc atctgcgcca cgaggtgatg
     2521 cagcagcatt gccgccgtgg gtttcctcgc aataagcccg gcccacgcct catgcgcttt
     2581 gcgttccgtt tgcacccagt gaccgggctt gttcttggct tgaatgccga tttctctgga
     2641 ctgcgtggcc atgcttatct ccatgcggta ggggtgccgc acggttgcgg caccatgcgc
     2701 aatcagctgc aacttttcgg cagcgcgaca acaattatgc gttgcgtaaa agtggcagtc
     2761 aattacagat tttctttaac ctacgcaatg agctattgcg gggggtgccg caatgagctg
     2821 ttgcgtaccc ccctttttta agttgttgat ttttaagtct ttcgcatttc gccctatatc
     2881 tagttctttg gtgcccaaag aagggcaccc ctgcggggtt cccccacgcc ttcggcgcgg
     2941 ctccccctcc ggcaaaaagt ggcccctccg gggcttgttg atcgactgcg cggccttcgg
     3001 ccttgcccaa ggtggcgctg cccccttgga acccccgcac tcgccgccgt gaggctcggg
     3061 gggcaggcgg gcgggcttcg cccttcgact gcccccactc gcataggctt gggtcgttcc
     3121 aggcgcgtca aggccaagcc gctgcgcggt cgctgcgcga gccttgaccc gccttccact
     3181 tggtgtccaa ccggcaagcg aagcgcgcag gccgcaggcc ggaggctttt ccccagagaa
     3241 aattaaaaaa attgatgggg caaggccgca ggccgcgcag ttggagccgg tgggtatgtg
     3301 gtcgaaggct gggtagccgg tgggcaatcc ctgtggtcaa gctcgtgggc aggcgcagcc
     3361 tgtccatcag cttgtccagc agggttgtcc acgggccgag cgaagcgagc cagccggtgg
     3421 ccgctcgcgg ccatcgtcca catatccacg ggctggcaag ggagcgcagc gaccgcgcag
     3481 ggcgaagccc ggagagcaag cccgtagggg gggcgcgccc agctgtctag ggcggcggat
     3541 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
     3601 tcgactgagc ctttcgtttt atttgatgcc t
//
