LOCUS       40924_39208        3521 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA114, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3521)
  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  JOURNAL   Unpublished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 3521)
  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 3521)
  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 3521)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3521)
  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            "Unpublished"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 JX560381.1.
FEATURES             Location/Qualifiers
     source          1..3521
                     /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"
     promoter        1749..1840
                     /label=AmpR promoter
     CDS             1841..2698
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      2705..2744
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            2891..2999
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      3010..3398
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     terminator      complement(3429..3515)
                     /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 ttaaattagt agcccgccta atgagcgggc ttttttttaa
     1741 ttcccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     1801 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagcattc agcattttcg
     1861 tgtggcgctg attccgtttt ttgcggcgtt ttgcctgccg gtgtttgcgc atccggaaac
     1921 cctggtgaaa gtgaaagatg cggaagatca actgggtgcg cgcgtgggct atattgaact
     1981 ggatctgaac agcggcaaaa ttctggaatc ttttcgtccg gaagaacgtt ttccgatgat
     2041 gagcaccttt aaagtgctgc tgtgcggtgc ggttctgagc cgtgtggatg cgggccagga
     2101 acaactgggc cgtcgtattc attatagcca gaacgatctg gtggaatata gcccggtgac
     2161 cgaaaaacat ctgaccgatg gcatgaccgt gcgtgaactg tgcagcgcgg cgattaccat
     2221 gagcgataac accgcggcga acctgctgct gacgaccatt ggcggtccga aagaactgac
     2281 cgcgtttctg cataacatgg gcgatcatgt gacccgtctg gatcgttggg aaccggaact
     2341 gaacgaagcg attccgaacg atgaacgtga taccaccatg ccggcagcaa tggcgaccac
     2401 cctgcgtaaa ctgctgacgg gtgagctgct gaccctggca agccgccagc aactgattga
     2461 ttggatggaa gcggataaag tggcgggtcc gctgctgcgt agcgcgctgc cggctggctg
     2521 gtttattgcg gataaaagcg gtgcgggcga acgtggcagc cgtggcatta ttgcggcgct
     2581 gggcccggat ggtaaaccga gccgtattgt ggtgatttat accaccggca gccaggcgac
     2641 gatggatgaa cgtaaccgtc agattgcgga aattggcgcg agcctgatta aacattggta
     2701 aaccgataca attaaaggct ccttttggag cctttttttt tggacgaccc ttgtcctttt
     2761 ccgctgcata accctgcttc ggggtcatta tagcgatttt ttcggtatat ccatcctttt
     2821 tcgcacgata tacaggattt tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac
     2881 ggcgtcagcc gggcaggata ggtgaagtag gcccacccgc gagcgggtgt tccttcttca
     2941 ctgtccctta ttcgcacctg gcggtgctca acgggaatcc tgctctgcga ggctggccgt
     3001 aggccggccg atctgaagat cagcagttca acctgttgat agtacgtact aagctctcat
     3061 gtttcacgta ctaagctctc atgtttaacg tactaagctc tcatgtttaa cgaactaaac
     3121 cctcatggct aacgtactaa gctctcatgg ctaacgtact aagctctcat gtttcacgta
     3181 ctaagctctc atgtttgaac aataaaatta atataaatca gcaacttaaa tagcctctaa
     3241 ggttttaagt tttataagaa aaaaaagaat atataaggct tttaaagcct ttaaggttta
     3301 acggttgtgg acaacaagcc agggatgtaa cgcactgaga agcccttaga gcctctcaaa
     3361 gcaattttga gtgacacagg aacacttaac ggctgacatg gggcgcgccc agctgtctag
     3421 ggcggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag
     3481 gcccagtctt tcgactgagc ctttcgtttt atttgatgcc t
//
