LOCUS       V004350                 6895 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004350
VERSION     V004350
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6895)
  AUTHORS   Kulman JD, Satake M, Harris JE.
  TITLE     A versatile system for site-specific enzymatic biotinylation and
            regulated expression of proteins in cultured mammalian cells
  JOURNAL   Protein Expr. Purif. 52 (2), 320-328 (2007)
   PUBMED   17084093
REFERENCE   2  (bases 1 to 6895)
  AUTHORS   Kulman JD, Satake M, Harris JE.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-APR-2006) Biochemistry, University of Washington, N.E.
            Pacific St., Seattle, WA 98195, USA
REFERENCE   3  (bases 1 to 6895)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6895)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein
            Expr. Purif."; date: "2007"; volume: "52"; issue: "2"; pages:
            "320-328"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (28-APR-2006) Biochemistry, University of Washington, N.E. Pacific
            St., Seattle, WA 98195, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    15..285
                     /label="tetracycline response element"
                     /note="contains seven copies of the tetracycline operator
                     tetO"
     promoter        318..356
                     /label="minimal CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     regulatory      477..485
                     /label="Kozak site"
                     /note="Kozak site"
                     /regulatory_class="ribosome_binding_site"
     regulatory      480..489
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    492..530
                     /label="biotin acceptor peptide (BioTag)-encoding"
                     /note="biotin acceptor peptide (BioTag)-encoding"
     misc_feature    532..551
                     /note="multiple cloning site; MCS"
     misc_feature    652..1238
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     regulatory      1270..1278
                     /gene="birA"
                     /label="Kozak site"
                     /note="Kozak site"
                     /regulatory_class="ribosome_binding_site"
     regulatory      1273..1282
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1282..1305
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             1318..2280
                     /label="BirA"
                     /note="E. coli biotin protein ligase"
     intron          2348..2920
                     /label="beta-globin intron"
                     /note="intron from rabbit beta-globin gene"
     CDS             2924..3046
                     /gene="HBB"
                     /label="Hemoglobin subunit beta"
                     /note="Hemoglobin subunit beta from Colobus guereza.
                     Accession#: Q9TT33"
     polyA_signal    3117..3172
                     /label="beta-globin poly(A) signal"
                     /note="rabbit beta-globin polyadenylation signal (Gil and
                     Proudfoot, 1987)"
     rep_origin      complement(3734..4322)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4496..5353)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(5354..5458)
                     /label="AmpR promoter"
     promoter        5563..5892
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             5985..6581
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     polyA_signal    6814..6895
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 ctcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc
       61 tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa
      121 gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc
      181 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
      241 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
      301 ctcggtaccc gggtcgaggt aggcgtgtac ggtgggaggc ctatataagc agagctcgtt
      361 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac
      421 accgggaccg atccagcctc cgcggccccg aattcgagct cggtacccgg ggatctgccg
      481 ccaccatggg cctgaacgac atcttcgaag ctcagaaaat cgaatggcac gggatccgat
      541 atcgcggccg cttgaacgac atttttgaag cgcaaaaaat tgaatggcat taaggccgcg
      601 ctagcctcga gaattcacgc gtcgagcatg catctagggc ggccaattcc gcccctctcc
      661 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
      721 ctatatgtga ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
      781 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
      841 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
      901 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
      961 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     1021 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
     1081 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt
     1141 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
     1201 tttcctttga aaaacacgat gataagcttg ccacaaccca aacttaagct tggtaccgag
     1261 ctcggatctg ccgccaccat ggattacaag gatgacgacg ataagagccc gagatccatg
     1321 aaggataaca ccgtgccact gaaattgatt gccctgttag cgaacggtga atttcactct
     1381 ggcgagcagt tgggtgaaac gctgggaatg agccgggcgg ctattaataa acacattcag
     1441 acactgcgtg actggggcgt tgatgtcttt accgttccgg gtaaaggata cagcctgcct
     1501 gagcctatcc agttacttaa tgctaaacag atattgggtc agctggatgg cggtagtgta
     1561 gccgtgctgc cagtgattga ctccacgaat cagtaccttc ttgatcgtat cggagagctt
     1621 aaatcgggcg atgcttgcat tgcagaatac cagcaggctg gccgtggtcg ccggggtcgg
     1681 aaatggtttt cgccttttgg cgcaaactta tatttgtcga tgttctggcg tctggaacaa
     1741 ggcccggcgg cggcgattgg tttaagtctg gttatcggta tcgtgatggc ggaagtatta
     1801 cgcaagctgg gtgcagataa agttcgtgtt aaatggccta atgacctcta tctgcaggat
     1861 cgcaagctgg caggcattct ggtggagctg actggcaaaa ctggcgatgc ggcgcaaata
     1921 gtcattggag ccgggatcaa catggcaatg cgccgtgttg aagagagtgt cgttaatcag
     1981 gggtggatca cgctgcagga agcggggatc aatctcgatc gtaatacgtt ggcggccatg
     2041 ctaatacgtg aattacgtgc tgcgttggaa ctcttcgaac aagaaggatt ggcaccttat
     2101 ctgtcgcgct gggaaaagct ggataatttt attaatcgcc cagtgaaact tatcattggt
     2161 gataaagaaa tatttggcat ttcacgcgga atagacaaac agggggcttt attacttgag
     2221 caggatggaa taataaaacc ctggatgggc ggtgaaatat ccctgcgtag tgcagaaaaa
     2281 taagaattca gatctctgca gctcgagtct agagggcccg tttatctcta gagctgagaa
     2341 cttcagggtg agtttgggga cccttgattg ttctttcttt ttcgctattg taaaattcat
     2401 gttatatgga gggggcaaag ttttcagggt gttgtttaga atgggaagat gtcccttgta
     2461 tcaccatgga ccctcatgat aattttgttt ctttcacttt ctactctgtt gacaaccatt
     2521 gtctcctctt attttctttt cattttctgt aacttttttc gttaaacttt agcttgcatt
     2581 tgtaacgaat ttttaaattc actttcgttt atttgtcaga ttgtaagtac tttctctaat
     2641 cacttttttt tcaaggcaat cagggtaatt atattgtact tcagcacagt tttagagaac
     2701 aattgttata attaaatgat aaggtagaat atttctgcat ataaattctg gctggcgtgg
     2761 aaatattctt attggtagaa acaactacat cctggtaatc atcctgcctt tctctttatg
     2821 gttacaatga tatacactgt ttgagatgag gataaaatac tctgagtcca aaccgggccc
     2881 ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt
     2941 tgttgtgctg tctcatcatt ttggcaaaga attcactcct caggtgcagg ctgcctatca
     3001 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc
     3061 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctgggtaata
     3121 aaggaaattt attttcattg caatagtgtg tgggaatttt ttgtgtctct cactcggaag
     3181 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc
     3241 aacatatgcc atatgctggc tgccatgaac aaaggtggct ataaagaggt catcagtata
     3301 tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga
     3361 ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac
     3421 atgttttact agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc
     3481 ttctcttatg aactcgactg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3541 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3601 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3661 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3721 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3781 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3841 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3901 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     3961 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4021 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4081 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4141 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     4201 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4261 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4321 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4381 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4441 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4501 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4561 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4621 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4681 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4741 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4801 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4861 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4921 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4981 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5041 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5101 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5161 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5221 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5281 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5341 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5401 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5461 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     5521 ataaaaatag gcgtatcacg aggccctttc gtcttcactc gagtgtgtca gttagggtgt
     5581 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5641 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5701 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5761 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5821 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5881 ggcttttgca aaaagcttgc atgcctgcag gtcggccgcc acgaccggtg ccgccaccat
     5941 cccctgaccc acgcccctga cccctcacaa ggagacgacc ttccatgacc gagtacaagc
     6001 ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg
     6061 cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg
     6121 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     6181 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     6241 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     6301 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     6361 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     6421 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     6481 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     6541 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc
     6601 cgcagcgccc gaccgaaagg agcgcacgac cccatggctc cgaccgaagc cgacccgggc
     6661 ggccccgccg accccgcacc cgcccccgag gcccaccgac tctagaggat cataatcagc
     6721 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     6781 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     6841 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgc
//
