LOCUS       40924_39493        5309 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA424, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5309)
  AUTHORS   Silva-Rocha R, Martinez-Garcia E, Calles B, Chavarria M, 
            Arce-Rodriguez A, de Las Heras A, Paez-Espino AD, Durante-Rodriguez 
            G, Kim J, Nikel PI, Platero R, de Lorenzo V.
  TITLE     The Standard European Vector Architecture (SEVA): a coherent 
            platform for the analysis and deployment of complex prokaryotic 
            phenotypes
  JOURNAL   Nucleic Acids Res. 41 (DATABASE ISSUE), D666-D675 (2013)
  PUBMED    23180763
REFERENCE   2  (bases 1 to 5309)
  AUTHORS   Silva-Rocha R, Martinez-Garcia E, de Lorenzo V.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-APR-2013) Systems Biology Program, CNB-CSIC, Darwin 3,
            Madrid, Madrid 28029, Spain
REFERENCE   3  (bases 1 to 5309)
  AUTHORS   Silva-Rocha R, Martinez-Garcia E, de Lorenzo V.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-SEP-2014) Systems Biology Program, CNB-CSIC, Darwin 3,
            Madrid, Madrid 28029, Spain
REFERENCE   4  (bases 1 to 5309)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5309)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 41 (DATABASE ISSUE), D666-D675 (2013)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (01-APR-2013) Systems Biology Program, CNB-CSIC, Darwin 3, Madrid, 
            Madrid 28029, Spain"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (30-SEP-2014) Systems Biology Program, CNB-CSIC, Darwin 3, Madrid, 
            Madrid 28029, Spain"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
            On Sep 30, 2014 this sequence version replaced KC847293.1.
FEATURES             Location/Qualifiers
     source          1..5309
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..88
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             89..1168
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        1403..1432
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    1440..1456
                     /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)."
     misc_feature    1483..1539
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1548..1571)
                     /label=F24
                     /note="F24"
     terminator      1582..1676
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1912..2700
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     oriT            2849..2957
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(3021..4166)
                     /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(4559..5090)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(5217..5303)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ttaattaatt gacaccatcg aatggtgcaa aacctttcgc ggtatggcat gatagcgccc
       61 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
      121 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
      181 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
      241 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
      301 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
      361 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
      421 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
      481 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
      541 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
      601 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
      661 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
      721 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca
      781 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct
      841 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt
      901 gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca
      961 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca
     1021 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc
     1081 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     1141 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagcgcg
     1201 aattgatctg gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca
     1261 ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt
     1321 cgctcaaggc gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg
     1381 gcaaatattc tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat
     1441 tgtgagcgga taacaatttc acaccctagg ccgcggccgc gcgaattcga gctcggtacc
     1501 cggggatcct ctagagtcga cctgcaggca tgcaagcttg cggccgcgtc gtgactggga
     1561 aaaccctggc gactagtctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc
     1621 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatccag
     1681 gggtccccaa taattacgat ttacgtattt aaatgaacct tgaccgaacg cagcggtggt
     1741 aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg
     1801 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc
     1861 agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat catgagggaa
     1921 gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat
     1981 ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag
     2041 ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg
     2101 cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc
     2161 cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct
     2221 aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag
     2281 ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt
     2341 gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt
     2401 gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag
     2461 cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg
     2521 ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc
     2581 atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca
     2641 gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa
     2701 taagacaatt gtccttttcc gctgcataac cctgcttcgg ggtcattata gcgatttttt
     2761 cggtatatcc atcctttttc gcacgatata caggattttg ccaaagggtt cgtgtagact
     2821 ttccttggtg tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga
     2881 gcgggtgttc cttcttcact gtcccttatt cgcacctggc ggtgctcaac gggaatcctg
     2941 ctctgcgagg ctggccgtag gccggccgcg atgcaggtgg ctgctgaacc cccagccgga
     3001 actgacccca caaggcccta gcgtttgcaa tgcaccaggt catcattgac ccaggcgtgt
     3061 tccaccaggc cgctgcctcg caactcttcg caggcttcgc cgacctgctc gcgccacttc
     3121 ttcacgcggg tggaatccga tccgcacatg aggcggaagg tttccagctt gagcgggtac
     3181 ggctcccggt gcgagctgaa atagtcgaac atccgtcggg ccgtcggcga cagcttgcgg
     3241 tacttctccc atatgaattt cgtgtagtgg tcgccagcaa acagcacgac gatttcctcg
     3301 tcgatcagga cctggcaacg ggacgttttc ttgccacggt ccaggacgcg gaagcggtgc
     3361 agcagcgaca ccgattccag gtgcccaacg cggtcggacg tgaagcccat cgccgtcgcc
     3421 tgtaggcgcg acaggcattc ctcggccttc gtgtaatacc ggccattgat cgaccagccc
     3481 aggtcctggc aaagctcgta gaacgtgaag gtgatcggct cgccgatagg ggtgcgcttc
     3541 gcgtactcca acacctgctg ccacaccagt tcgtcatcgt cggcccgcag ctcgacgccg
     3601 gtgtaggtga tcttcacgtc cttgttgacg tggaaaatga ccttgttttg cagcgcctcg
     3661 cgcgggattt tcttgttgcg cgtggtgaac agggcagagc gggccgtgtc gtttggcatc
     3721 gctcgcatcg tgtccggcca cggcgcaata tcgaacaagg aaagctgcat ttccttgatc
     3781 tgctgcttcg tgtgtttcag caacgcggcc tgcttggctt cgctgacctg ttttgccagg
     3841 tcctcgccgg cggtttttcg cttcttggtc gtcatagttc ctcgcgtgtc gatggtcatc
     3901 gacttcgcca aacctgccgc ctcctgttcg agacgacgcg aacgctccac ggcggccgat
     3961 ggcgcgggca gggcaggggg agccagttgc acgctgtcgc gctcgatctt ggccgtagct
     4021 tgctggacta tcgagccgac ggactggaag gtttcgcggg gcgcacgcat gacggtgcgg
     4081 cttgcgatgg tttcggcatc ctcggcggaa aaccccgcgt cgatcagttc ttgcctgtat
     4141 gccttccggt caaacgtccg attcattcac cctccttgcg ggattgcccc ggaattaatt
     4201 ccccggatcg atccgtcgat cttgatcccc tgcgccatca gatccttggc ggcaagaaag
     4261 ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca gctggcaatt
     4321 ccggttcgct tgctgtccat aaaaccgccc agtctagcta tcgccatgta agcccactgc
     4381 aagctacctg ctttctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga
     4441 cattcatccg gggtcagcac cgtttctgcg gactggcttt ctacgtggct gccatttttg
     4501 gggtgaggcc gttcgcggcc gaggggcgca gcccctgggg ggatgggagg cccgcgttag
     4561 cgggccggga gggttcgaga agggggggca ccccccttcg gcgtgcgcgg tcacgcgcac
     4621 agggcgcagc cctggttaaa aacaaggttt ataaatattg gtttaaaagc aggttaaaag
     4681 acaggttagc ggtggccgaa aaacgggcgg aaacccttgc aaatgctgga ttttctgcct
     4741 gtggacagcc cctcaaatgt caataggtgc gcccctcatc tgtcagcact ctgcccctca
     4801 agtgtcaagg atcgcgcccc tcatctgtca gtagtcgcgc ccctcaagtg tcaataccgc
     4861 agggcactta tccccaggct tgtccacatc atctgtggga aactcgcgta aaatcaggcg
     4921 ttttcgccga tttgcgaggc tggccagctc cacgtcgccg gccgaaatcg agcctgcccc
     4981 tcatctgtca acgccgcgcc gggtgagtcg gcccctcaag tgtcaacgtc cgcccctcat
     5041 ctgtcagtga gggccaagtt ttccgcgagg tatccacaac gccggcggcc ctacatggct
     5101 ctgctgtagt gagtgggttg cgctccggca gcggtcctga tcccccgcag aaaaaaagga
     5161 tctcaagaag atcctttgat cttttctacg gcgcgcccag ctgtctaggg cggcggattt
     5221 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc
     5281 gactgagcct ttcgttttat ttgatgcct
//
