LOCUS       V008681                 6834 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V008681
VERSION     V008681
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6834)
  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 6834)
  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 6834)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6834)
  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..6834
                     /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"
     misc_feature    471..490
                     /note="multiple cloning site; MCS"
     misc_feature    491..529
                     /label="biotin acceptor peptide (BioTag)-encoding"
                     /note="biotin acceptor peptide (BioTag)-encoding"
     misc_feature    591..1177
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     regulatory      1209..1217
                     /gene="birA"
                     /label="Kozak site"
                     /note="Kozak site"
                     /regulatory_class="ribosome_binding_site"
     regulatory      1212..1221
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1221..1244
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             1257..2219
                     /label="BirA"
                     /note="E. coli biotin protein ligase"
     intron          2287..2859
                     /label="beta-globin intron"
                     /note="intron from rabbit beta-globin gene"
     CDS             2863..2985
                     /gene="HBB"
                     /label="Hemoglobin subunit beta"
                     /note="Hemoglobin subunit beta from Colobus guereza.
                     Accession#: Q9TT33"
     polyA_signal    3056..3111
                     /label="beta-globin poly(A) signal"
                     /note="rabbit beta-globin polyadenylation signal (Gil and
                     Proudfoot, 1987)"
     rep_origin      complement(3673..4261)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4435..5292)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(5293..5397)
                     /label="AmpR promoter"
     promoter        5502..5831
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             5924..6520
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     polyA_signal    6753..6834
                     /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 ggatccgata
      481 tcgcggccgc ttgaacgaca tttttgaagc gcaaaaaatt gaatggcatt aaggccgcgc
      541 tagcctcgag aattcacgcg tcgagcatgc atctagggcg gccaattccg cccctctccc
      601 tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
      661 tatatgtgat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
      721 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
      781 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
      841 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
      901 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
      961 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
     1021 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
     1081 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
     1141 ttcctttgaa aaacacgatg ataagcttgc cacaacccaa acttaagctt ggtaccgagc
     1201 tcggatctgc cgccaccatg gattacaagg atgacgacga taagagcccg agatccatga
     1261 aggataacac cgtgccactg aaattgattg ccctgttagc gaacggtgaa tttcactctg
     1321 gcgagcagtt gggtgaaacg ctgggaatga gccgggcggc tattaataaa cacattcaga
     1381 cactgcgtga ctggggcgtt gatgtcttta ccgttccggg taaaggatac agcctgcctg
     1441 agcctatcca gttacttaat gctaaacaga tattgggtca gctggatggc ggtagtgtag
     1501 ccgtgctgcc agtgattgac tccacgaatc agtaccttct tgatcgtatc ggagagctta
     1561 aatcgggcga tgcttgcatt gcagaatacc agcaggctgg ccgtggtcgc cggggtcgga
     1621 aatggttttc gccttttggc gcaaacttat atttgtcgat gttctggcgt ctggaacaag
     1681 gcccggcggc ggcgattggt ttaagtctgg ttatcggtat cgtgatggcg gaagtattac
     1741 gcaagctggg tgcagataaa gttcgtgtta aatggcctaa tgacctctat ctgcaggatc
     1801 gcaagctggc aggcattctg gtggagctga ctggcaaaac tggcgatgcg gcgcaaatag
     1861 tcattggagc cgggatcaac atggcaatgc gccgtgttga agagagtgtc gttaatcagg
     1921 ggtggatcac gctgcaggaa gcggggatca atctcgatcg taatacgttg gcggccatgc
     1981 taatacgtga attacgtgct gcgttggaac tcttcgaaca agaaggattg gcaccttatc
     2041 tgtcgcgctg ggaaaagctg gataatttta ttaatcgccc agtgaaactt atcattggtg
     2101 ataaagaaat atttggcatt tcacgcggaa tagacaaaca gggggcttta ttacttgagc
     2161 aggatggaat aataaaaccc tggatgggcg gtgaaatatc cctgcgtagt gcagaaaaat
     2221 aagaattcag atctctgcag ctcgagtcta gagggcccgt ttatctctag agctgagaac
     2281 ttcagggtga gtttggggac ccttgattgt tctttctttt tcgctattgt aaaattcatg
     2341 ttatatggag ggggcaaagt tttcagggtg ttgtttagaa tgggaagatg tcccttgtat
     2401 caccatggac cctcatgata attttgtttc tttcactttc tactctgttg acaaccattg
     2461 tctcctctta ttttcttttc attttctgta acttttttcg ttaaacttta gcttgcattt
     2521 gtaacgaatt tttaaattca ctttcgttta tttgtcagat tgtaagtact ttctctaatc
     2581 actttttttt caaggcaatc agggtaatta tattgtactt cagcacagtt ttagagaaca
     2641 attgttataa ttaaatgata aggtagaata tttctgcata taaattctgg ctggcgtgga
     2701 aatattctta ttggtagaaa caactacatc ctggtaatca tcctgccttt ctctttatgg
     2761 ttacaatgat atacactgtt tgagatgagg ataaaatact ctgagtccaa accgggcccc
     2821 tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt
     2881 gttgtgctgt ctcatcattt tggcaaagaa ttcactcctc aggtgcaggc tgcctatcag
     2941 aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc
     3001 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tgggtaataa
     3061 aggaaattta ttttcattgc aatagtgtgt gggaattttt tgtgtctctc actcggaagg
     3121 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     3181 acatatgcca tatgctggct gccatgaaca aaggtggcta taaagaggtc atcagtatat
     3241 gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat
     3301 tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca
     3361 tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct
     3421 tctcttatga actcgactgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3481 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3541 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3601 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3661 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3721 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3781 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3841 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     3901 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3961 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4021 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4081 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     4141 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4201 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4261 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4321 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4381 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4441 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4501 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4561 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4621 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4681 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4741 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4801 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4861 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4921 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4981 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5041 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5101 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5161 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5221 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5281 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5341 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5401 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5461 taaaaatagg cgtatcacga ggccctttcg tcttcactcg agtgtgtcag ttagggtgtg
     5521 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5581 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5641 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     5701 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5761 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5821 gcttttgcaa aaagcttgca tgcctgcagg tcggccgcca cgaccggtgc cgccaccatc
     5881 ccctgaccca cgcccctgac ccctcacaag gagacgacct tccatgaccg agtacaagcc
     5941 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc
     6001 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt
     6061 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     6121 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     6181 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     6241 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     6301 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     6361 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     6421 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     6481 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc
     6541 gcagcgcccg accgaaagga gcgcacgacc ccatggctcc gaccgaagcc gacccgggcg
     6601 gccccgccga ccccgcaccc gcccccgagg cccaccgact ctagaggatc ataatcagcc
     6661 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     6721 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     6781 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgc
//
