LOCUS       40924_39538        4078 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA443, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4078)
  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  JOURNAL   Unpublished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 4078)
  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 4078)
  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 4078)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 4078)
  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            "Unpublished"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 JX560366.1.
FEATURES             Location/Qualifiers
     source          1..4078
                     /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    249..305
                     /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"
     rep_origin      2073..2661
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2718..3548)
                     /codon_start=1
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
                     /translation="VASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     rep_origin      3562..3913
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     terminator      complement(3986..4072)
                     /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 atgattacga attcgagctc ggtacccggg gatcctctag agtcgacctg caggcatgca
      301 agcttggcac 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 aggccggccg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     2041 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2101 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2161 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2221 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2281 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2341 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2401 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2461 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggca tccggtaagc
     2521 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2581 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2641 ggggggcgga gcctatggaa aaacgccagc aacgcggccg tgaaaggcag gccggtccgt
     2701 ggtggccacg gcctctaggc cagatccagc ggcatctggg ttagtcgagc gcgggccgct
     2761 tcccatgtct caccagggcg agcctgtttc gcgatctcag catctgaaat cttcccggcc
     2821 ttgcgcttcg ctggggcctt acccaccgcc ttggcgggct tcttcggtcc aaaactgaac
     2881 aacagatgtg tgaccttgcg cccggtcttt cgctgcgccc actccacctg tagcgggctg
     2941 tgctcgttga tctgcgtcac ggctggatca agcactcgca acttgaagtc cttgatcgag
     3001 ggataccggc cttccagttg aaaccacttt cgcagctggt caatttctat ttcgcgctgg
     3061 ccgatgctgt cccattgcat gagcagctcg taaagcctga tcgcgtgggt gctgtccatc
     3121 ttggccacgt cagccaaggc gtatttggtg aactgtttgg tgagttccgt caggtacggc
     3181 agcatgtctt tggtgaacct gagttctaca cggccctcac cctcccggta gatgattgtt
     3241 tgcacccagc cggtaatcat cacactcggt cttttcccct tgccattggg ctcttgggtt
     3301 aaccggactt cccgccgttt caggcgcagg gccgcttctt tgagctggtt gtaggaagat
     3361 tcgataggga cacccgccat cgtcgctatg tcctccgccg tcactgaata catcacttca
     3421 tcggtgacag gctcgctcct cttcacctgg ctaatacagg ccagaacgat ccgctgttcc
     3481 tgaacactga ggcgatacgc ggcctcgacc agggcattgc ttttgtaaac cattgggggt
     3541 gaggccacgt tcgacattcc ttgtgtataa ggggacactg tatctgcgtc ccacaataca
     3601 acaaatccgt ccctttacaa caacaaatcc gtcccttctt aacaacaaat ccgtccctta
     3661 atggcaacaa atccgtccct ttttaaactc tacaggccac ggattacgtg gcctgtagac
     3721 gtcctaaaag gtttaaaagg gaaaaggaag aaaagggtgg aaacgcaaaa aacgcaccac
     3781 tacgtggccc cgttggggcc gcatttgtgc ccctgaaggg gcgggggagg cgtctgggca
     3841 atccccgttt taccagtccc ctatcgccgc ctgagagggc gcaggaagcg agtaatcagg
     3901 gtatcgaggc ggattcaccc ttggcgtcca accagcggca ccagcggcgc ctgagagggg
     3961 cgcgcccagc tgtctagggc ggcggatttg tcctactcag gagagcgttc accgacaaac
     4021 aacagataaa acgaaaggcc cagtctttcg actgagcctt tcgttttatt tgatgcct
//
