LOCUS       40924_39588        4356 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA543, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4356)
  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  JOURNAL   Unpublished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 4356)
  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 4356)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4356)
  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            "Unpublished"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..4356
                     /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             813..2000
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     oriT            2149..2257
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      2351..2939
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2996..3826)
                     /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      3840..4191
                     /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(4264..4350)
                     /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 ttaaatttga cagcttatca tcgataaact gtaatgcggt agtttatcac
      781 agttaaattg ctaacgcagt caggcaccgt gtatgaaatc taacaatgcg ctcatcgtca
      841 tcctcggcac cgtcaccctg gatgctgtag gcataggctt ggttatgccg gtactgccgg
      901 gcctcttgcg ggatatcgtc cattccgaca gcatcgccag tcactatggc gtgctgctag
      961 cgctatatgc gttgatgcaa tttctatgcg cacccgttct cggagcactg tccgaccgct
     1021 ttggccgccg cccagtcctg ctcgcttcgc tacttggagc cactatcgac tacgcgatca
     1081 tggcgaccac acccgtcctg tggattctct acgccggacg catcgtggcc ggcatcaccg
     1141 gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg
     1201 ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg
     1261 ccgggggact gttgggcgcc atctccctgc acgcaccatt ccttgcggcg gcggtgctca
     1321 acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgcc
     1381 gtccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     1441 ccattgtggc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     1501 cagcgctctg ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc
     1561 tgtcgcttgc ggtattcgga atcttgcacg ccctcgctca agccttcgtc actggtcccg
     1621 ccaccaaacg tttcggcgag aagcaggcca ttatcgccgg catggcggcc gacgcgctgg
     1681 gctacgtctt gctggcgttc gcgacgcgag gctggatggc cttccccatt atgattcttc
     1741 tcgcttccgg cggcatcggg atgcccgcgt tgcaggccat gctgtccagg caggtagatg
     1801 acgaccatca gggacagctt caaggatcgc tcgcggctct taccagccta acttcgatca
     1861 ctggaccgct gatcgtcacg gcgatttatg ccgcctcggc gagcacatgg aacgggttgg
     1921 catggattgt aggcgccgcc ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat
     1981 ggagccgggc cacctcgacc tgagacaatt gtccttttcc gctgcataac cctgcttcgg
     2041 ggtcattata gcgatttttt cggtatatcc atcctttttc gcacgatata caggattttg
     2101 ccaaagggtt cgtgtagact ttccttggtg tatccaacgg cgtcagccgg gcaggatagg
     2161 tgaagtaggc ccacccgcga gcgggtgttc cttcttcact gtcccttatt cgcacctggc
     2221 ggtgctcaac gggaatcctg ctctgcgagg ctggccgtag gccggccgat aatctcatga
     2281 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2341 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2401 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2461 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     2521 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2581 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     2641 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     2701 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     2761 ttcccgaagg gagaaaggcg gacaggcatc cggtaagcgg cagggtcgga acaggagagc
     2821 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2881 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     2941 acgccagcaa cgcggccgtg aaaggcaggc cggtccgtgg tggccacggc ctctaggcca
     3001 gatccagcgg catctgggtt agtcgagcgc gggccgcttc ccatgtctca ccagggcgag
     3061 cctgtttcgc gatctcagca tctgaaatct tcccggcctt gcgcttcgct ggggccttac
     3121 ccaccgcctt ggcgggcttc ttcggtccaa aactgaacaa cagatgtgtg accttgcgcc
     3181 cggtctttcg ctgcgcccac tccacctgta gcgggctgtg ctcgttgatc tgcgtcacgg
     3241 ctggatcaag cactcgcaac ttgaagtcct tgatcgaggg ataccggcct tccagttgaa
     3301 accactttcg cagctggtca atttctattt cgcgctggcc gatgctgtcc cattgcatga
     3361 gcagctcgta aagcctgatc gcgtgggtgc tgtccatctt ggccacgtca gccaaggcgt
     3421 atttggtgaa ctgtttggtg agttccgtca ggtacggcag catgtctttg gtgaacctga
     3481 gttctacacg gccctcaccc tcccggtaga tgattgtttg cacccagccg gtaatcatca
     3541 cactcggtct tttccccttg ccattgggct cttgggttaa ccggacttcc cgccgtttca
     3601 ggcgcagggc cgcttctttg agctggttgt aggaagattc gatagggaca cccgccatcg
     3661 tcgctatgtc ctccgccgtc actgaataca tcacttcatc ggtgacaggc tcgctcctct
     3721 tcacctggct aatacaggcc agaacgatcc gctgttcctg aacactgagg cgatacgcgg
     3781 cctcgaccag ggcattgctt ttgtaaacca ttgggggtga ggccacgttc gacattcctt
     3841 gtgtataagg ggacactgta tctgcgtccc acaatacaac aaatccgtcc ctttacaaca
     3901 acaaatccgt cccttcttaa caacaaatcc gtcccttaat ggcaacaaat ccgtcccttt
     3961 ttaaactcta caggccacgg attacgtggc ctgtagacgt cctaaaaggt ttaaaaggga
     4021 aaaggaagaa aagggtggaa acgcaaaaaa cgcaccacta cgtggccccg ttggggccgc
     4081 atttgtgccc ctgaaggggc gggggaggcg tctgggcaat ccccgtttta ccagtcccct
     4141 atcgccgcct gagagggcgc aggaagcgag taatcagggt atcgaggcgg attcaccctt
     4201 ggcgtccaac cagcggcacc agcggcgcct gagaggggcg cgcccagctg tctagggcgg
     4261 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     4321 gtctttcgac tgagcctttc gttttatttg atgcct
//
