LOCUS       40924_39258        3420 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA214, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3420)
  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  JOURNAL   Unpublished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 3420)
  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 3420)
  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 (25-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 3420)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3420)
  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            "Unpublished"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 
            (25-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 25, 2014 this sequence version replaced JX560382.1.
FEATURES             Location/Qualifiers
     source          1..3420
                     /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             1808..2620
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            2790..2898
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      2909..3297
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     terminator      complement(3328..3414)
                     /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 ttaaatttgt gtctcaaaat ctctgatgtt acattgcaca
     1741 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     1801 gggtgttatg agccatattc agcgtgaaac gagctgtagc cgtccgcgtc tgaacagcaa
     1861 catggatgcg gatctgtatg gctataaatg ggcgcgtgat aacgtgggtc agagcggcgc
     1921 gaccatttat cgtctgtatg gcaaaccgga tgcgccggaa ctgtttctga aacatggcaa
     1981 aggcagcgtg gcgaacgatg tgaccgatga aatggtgcgt ctgaactggc tgaccgaatt
     2041 tatgccgctg ccgaccatta aacattttat tcgcaccccg gatgatgcgt ggctgctgac
     2101 caccgcgatt ccgggcaaaa ccgcgtttca ggtgctggaa gaatatccgg atagcggcga
     2161 aaacattgtg gatgcgctgg ccgtgtttct gcgtcgtctg catagcattc cggtgtgcaa
     2221 ctgcccgttt aacagcgatc gtgtgtttcg tctggcccag gcgcagagcc gtatgaacaa
     2281 cggcctggtg gatgcgagcg attttgatga tgaacgtaac ggctggccgg tggaacaggt
     2341 gtggaaagaa atgcataaac tgctgccgtt tagcccggat agcgtggtga cccacggcga
     2401 ttttagcctg gataacctga ttttcgatga aggcaaactg attggctgca ttgatgtggg
     2461 ccgtgtgggc attgcggatc gttatcagga tctggccatt ctgtggaact gcctgggcga
     2521 atttagcccg agcctgcaaa aacgtctgtt tcagaaatat ggcattgata atccggatat
     2581 gaacaaactg caatttcatc tgatgctgga tgaatttttc taataattaa ttggaccgcg
     2641 gtccgcgcgt tgtccttttc cgctgcataa ccctgcttcg gggtcattat agcgattttt
     2701 tcggtatatc catccttttt cgcacgatat acaggatttt gccaaagggt tcgtgtagac
     2761 tttccttggt gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg
     2821 agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct
     2881 gctctgcgag gctggccgta ggccggccga tctgaagatc agcagttcaa cctgttgata
     2941 gtacgtacta agctctcatg tttcacgtac taagctctca tgtttaacgt actaagctct
     3001 catgtttaac gaactaaacc ctcatggcta acgtactaag ctctcatggc taacgtacta
     3061 agctctcatg tttcacgtac taagctctca tgtttgaaca ataaaattaa tataaatcag
     3121 caacttaaat agcctctaag gttttaagtt ttataagaaa aaaaagaata tataaggctt
     3181 ttaaagcctt taaggtttaa cggttgtgga caacaagcca gggatgtaac gcactgagaa
     3241 gcccttagag cctctcaaag caattttgag tgacacagga acacttaacg gctgacatgg
     3301 ggcgcgccca gctgtctagg gcggcggatt tgtcctactc aggagagcgt tcaccgacaa
     3361 acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct
//
