LOCUS       V006454                 6744 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V006454
VERSION     V006454
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6744)
  AUTHORS   Paetzold B, Carolis C, Ferrar T, Serrano L, Lluch-Senar M.
  TITLE     In situ overlap and sequence synthesis during DNA assembly
  JOURNAL   ACS Synth Biol 2 (12), 750-755 (2013)
   PUBMED   24161008
REFERENCE   2  (bases 1 to 6744)
  AUTHORS   Paetzold B.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-MAR-2013) EMBL-CRG Systems Biology Research Unit,
            Centre for Genomic Regulation, Dr. Aiguader 88, Barcelona 08003,
            Spain
REFERENCE   3  (bases 1 to 6744)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6744)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
            Biol"; date: "2013"; volume: "2"; issue: "12"; pages: "750-755"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (21-MAR-2013) EMBL-CRG Systems Biology Research Unit, Centre for
            Genomic Regulation, Dr. Aiguader 88, Barcelona 08003, Spain"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        216..234
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    235..259
                     /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)."
     RBS             274..296
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             309..326
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             336..359
                     /label="HRV 3C site"
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases"
     CDS             360..1760
                     /gene="dnaA"
                     /label="Chromosomal replication initiator protein DnaA"
                     /note="Chromosomal replication initiator protein DnaA from
                     Escherichia coli (strain K12). Accession#: P03004"
     CDS             1820..1837
                     /label="6xHis"
                     /note="6xHis affinity tag"
     terminator      1904..1951
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     rep_origin      1988..2443
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(2539..3351)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      3473..4061
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(4247..4389)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(4494..4682)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(5457..5478)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(5494..6573)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(6574..6651)
                     /label="lacI promoter"
ORIGIN
        1 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
       61 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
      121 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
      181 cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt
      241 gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata
      301 ccatgaaaca tcaccatcac catcactccg cgggtctgga agttctgttc caggggcccg
      361 tgtcactttc gctttggcag cagtgtcttg cccgattgca ggatgagtta ccagccacag
      421 aattcagtat gtggatacgc ccattgcagg cggaactgag cgataacacg ctggccctgt
      481 acgcgccaaa ccgttttgtc ctcgattggg tacgggacaa gtaccttaat aatatcaatg
      541 gactgctaac cagtttctgc ggagcggatg ccccacagct gcgttttgaa gtcggcacca
      601 aaccggtgac gcaaacgcca caagcggcag tgacgagcaa cgtcgcggcc cctgcacagg
      661 tggcgcaaac gcagccgcaa cgtgctgcgc cttctacgcg ctcaggttgg gataacgtcc
      721 cggccccggc agaaccgacc tatcgttcta acgtaaacgt caaacacacg tttgataact
      781 tcgttgaagg taaatctaac caactggcgc gcgcggcggc tcgccaggtg gcggataacc
      841 ctggcggtgc ctataacccg ttgttccttt atggcggcac gggtctgggt aaaactcacc
      901 tgctgcatgc ggtgggtaac ggcattatgg cgcgcaagcc gaatgccaaa gtggtttata
      961 tgcactccga gcgctttgtt caggacatgg ttaaagccct gcaaaacaac gcgatcgaag
     1021 agtttaaacg ctactaccgt tccgtagatg cactgctgat cgacgatatt cagttttttg
     1081 ctaataaaga acgatctcag gaagagtttt tccacacctt caacgccctg ctggaaggta
     1141 atcaacagat cattctcacc tcggatcgct atccgaaaga gatcaacggc gttgaggatc
     1201 gtttgaaatc ccgcttcggt tggggactga ctgtggcgat cgaaccgcca gagctggaaa
     1261 cccgtgtggc gatcctgatg aaaaaggccg acgaaaacga cattcgtttg ccgggcgaag
     1321 tggcgttctt tatcgccaag cgtctacgat ctaacgtacg tgagctggaa ggggcgctga
     1381 accgcgtcat tgccaatgcc aactttaccg gacgggcgat caccatcgac ttcgtgcgtg
     1441 aggcgctgcg cgacttgctg gcattgcagg aaaaactggt caccatcgac aatattcaga
     1501 agacggtggc ggagtactac aagatcaaag tcgcggatct cctttccaag cgtcgatccc
     1561 gctcggtggc gcgtccgcgc cagatggcga tggcgctggc gaaagagctg actaaccaca
     1621 gtctgccgga gattggcgat gcgtttggtg gccgtgacca cacgacggtg cttcatgcct
     1681 gccgtaagat cgagcagttg cgtgaagaga gccacgatat taaagaagat ttttcaaatt
     1741 taatcagaac attgtcatcg taacatatgc gccattaacc tgatgttctg gggaatataa
     1801 gcttgcggcc gcactcgagc accaccacca ccaccactga gatccggctg ctaacaaagc
     1861 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     1921 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc
     1981 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2041 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2101 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     2161 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     2221 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     2281 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     2341 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     2401 aaatttaacg cgaattttaa caaaatatta acgtttacaa tttcaggtgg cacttttcgg
     2461 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2521 ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     2581 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     2641 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     2701 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     2761 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
     2821 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     2881 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     2941 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     3001 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     3061 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     3121 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     3181 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     3241 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     3301 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct
     3361 tgtattactg tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg
     3421 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3481 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3541 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3601 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3661 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3721 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3781 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3841 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3901 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3961 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4021 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4081 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4141 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4201 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     4261 ttctccttac gcatctgtgc ggtatttcac accgcatata tggtgcactc tcagtacaat
     4321 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     4381 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4441 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4501 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     4561 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     4621 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     4681 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     4741 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     4801 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     4861 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     4921 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     4981 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     5041 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     5101 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg
     5161 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     5221 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     5281 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     5341 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     5401 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     5461 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5521 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5581 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     5641 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     5701 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     5761 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     5821 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     5881 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     5941 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     6001 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     6061 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     6121 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     6181 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     6241 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     6301 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     6361 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     6421 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     6481 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     6541 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     6601 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     6661 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     6721 tgagcaccgc cgccgcaagg aatg
//
