LOCUS       40924_39433        3273 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA314, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3273)
  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  JOURNAL   Unpublished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 3273)
  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 3273)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3273)
  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            "Unpublished"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..3273
                     /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=R24
                     /note="R24"
     terminator      1582..1676
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        1724..1826
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             1827..2483
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     oriT            2635..2743
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      2762..3150
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     terminator      complement(3181..3267)
                     /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 aattaatgtg 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 ttaaattggc gaaaatgaga cgttgatcgg cacgtaagag
     1741 gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga
     1801 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
     1861 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
     1921 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
     1981 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
     2041 aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt
     2101 acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg
     2161 atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg
     2221 cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga
     2281 gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca
     2341 ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc
     2401 atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact
     2461 gcgatgagtg gcagggcggg gcgtaatttg acttttgtcc ttttccgctg cataaccctg
     2521 cttcggggtc attatagcga ttttttcggt atatccatcc tttttcgcac gatatacagg
     2581 attttgccaa agggttcgtg tagactttcc ttggtgtatc caacggcgtc agccgggcag
     2641 gataggtgaa gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca
     2701 cctggcggtg ctcaacggga atcctgctct gcgaggctgg ccgtaggccg gccggcgcgc
     2761 cgatctgaag atcagcagtt caacctgttg atagtacgta ctaagctctc atgtttcacg
     2821 tactaagctc tcatgtttaa cgtactaagc tctcatgttt aacgaactaa accctcatgg
     2881 ctaacgtact aagctctcat ggctaacgta ctaagctctc atgtttcacg tactaagctc
     2941 tcatgtttga acaataaaat taatataaat cagcaactta aatagcctct aaggttttaa
     3001 gttttataag aaaaaaaaga atatataagg cttttaaagc ctttaaggtt taacggttgt
     3061 ggacaacaag ccagggatgt aacgcactga gaagccctta gagcctctca aagcaatttt
     3121 gagtgacaca ggaacactta acggctgaca tggggcgcgc ccagctgtct agggcggcgg
     3181 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
     3241 tttcgactga gcctttcgtt ttatttgatg cct
//
