LOCUS       V004349                 7032 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004349
VERSION     V004349
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7032)
  AUTHORS   Mize GJ, Harris JE, Takayama TK, Kulman JD.
  TITLE     Regulated expression of active biotinylated G-protein coupled
            receptors in mammalian cells
  JOURNAL   Protein Expr. Purif. 57 (2), 280-289 (2008)
   PUBMED   18042400
REFERENCE   2  (bases 1 to 7032)
  AUTHORS   Mize GJ, Harris JE, Takayama TK, Kulman JD.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-AUG-2007) Department of Biochemistry, University of
            Washington, N.E. Pacific Street, Seattle, WA 98195, USA
REFERENCE   3  (bases 1 to 7032)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7032)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein
            Expr. Purif."; date: "2008"; volume: "57"; issue: "2"; pages:
            "280-289"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (04-AUG-2007) Department of Biochemistry, University of Washington,
            N.E. Pacific Street, Seattle, WA 98195, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7032
                     /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"
     misc_feature    570..608
                     /label="biotin acceptor peptide (BioTag)-encoding"
                     /note="biotin acceptor peptide (BioTag)-encoding"
     misc_feature    609..628
                     /note="multiple cloning site; MCS"
     misc_feature    729..1315
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     regulatory      1326..1334
                     /label="Kozak site"
                     /note="Kozak site"
                     /regulatory_class="ribosome_binding_site"
     CDS             1335..1424
                     /label="prolactin signal sequence"
                     /note="signal sequence from bovine prolactin"
     CDS             1425..1448
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             1455..2417
                     /label="BirA"
                     /note="E. coli biotin protein ligase"
     intron          2485..3057
                     /label="beta-globin intron"
                     /note="intron from rabbit beta-globin gene"
     CDS             3061..3183
                     /gene="HBB"
                     /label="Hemoglobin subunit beta"
                     /note="Hemoglobin subunit beta from Colobus guereza.
                     Accession#: Q9TT33"
     polyA_signal    3254..3309
                     /label="beta-globin poly(A) signal"
                     /note="rabbit beta-globin polyadenylation signal (Gil and
                     Proudfoot, 1987)"
     rep_origin      complement(3871..4459)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4633..5490)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(5491..5595)
                     /label="AmpR promoter"
     promoter        5700..6029
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             6122..6718
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     polyA_signal    6951..7032
                     /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 ccaccatgcc gtcttctgtc tcgtggggca tcctcctgct ggcaggcctg tgctgcctgg
      541 tccctgtctc cctggctgag cgtggatctt tgaacgacat ttttgaagcg caaaaaattg
      601 aatggcatgg atccgatatc gcggccgctt gaacgacatt tttgaagcgc aaaaaattga
      661 atggcattaa ggccgcgcta gcctcgagaa ttcacgcgtc gagcatgcat ctagggcggc
      721 caattccgcc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
      781 ggccggtgtg cgtttgtcta tatgtgattt tccaccatat tgccgtcttt tggcaatgtg
      841 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
      901 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
      961 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     1021 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     1081 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     1141 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     1201 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     1261 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aagcttgcca caacccaaac
     1321 ttaaggccgc caccatggac agcaaaggtt cgtcgcagaa agggtcccgc ctgctcctgc
     1381 tgctggtggt gtcaaatcta ctcttgtgcc agggtgtggt ctccgattat aaggatgatg
     1441 atgataagct taagatgaag gataacaccg tgccactgaa attgattgcc ctgttagcga
     1501 acggtgaatt tcactctggc gagcagttgg gtgaaacgct gggaatgagc cgggcggcta
     1561 ttaataaaca cattcagaca ctgcgtgact ggggcgttga tgtctttacc gttccgggta
     1621 aaggatacag cctgcctgag cctatccagt tacttaatgc taaacagata ttgggtcagc
     1681 tggatggcgg tagtgtagcc gtgctgccag tgattgactc cacgaatcag taccttcttg
     1741 atcgtatcgg agagcttaaa tcgggcgatg cttgcattgc agaataccag caggctggcc
     1801 gtggtcgccg gggtcggaaa tggttttcgc cttttggcgc aaacttatat ttgtcgatgt
     1861 tctggcgtct ggaacaaggc ccggcggcgg cgattggttt aagtctggtt atcggtatcg
     1921 tgatggcgga agtattacgc aagctgggtg cagataaagt tcgtgttaaa tggcctaatg
     1981 acctctatct gcaggatcgc aagctggcag gcattctggt ggagctgact ggcaaaactg
     2041 gcgatgcggc gcaaatagtc attggagccg ggatcaacat ggcaatgcgc cgtgttgaag
     2101 agagtgtcgt taatcagggg tggatcacgc tgcaggaagc ggggatcaat ctcgatcgta
     2161 atacgttggc ggccatgcta atacgtgaat tacgtgctgc gttggaactc ttcgaacaag
     2221 aaggattggc accttatctg tcgcgctggg aaaagctgga taattttatt aatcgcccag
     2281 tgaaacttat cattggtgat aaagaaatat ttggcatttc acgcggaata gacaaacagg
     2341 gggctttatt acttgagcag gatggaataa taaaaccctg gatgggcggt gaaatatccc
     2401 tgcgtagtgc agaaaaataa gaattcagat ctctgcagct cgagtctaga gggcccgttt
     2461 atctctagag ctgagaactt cagggtgagt ttggggaccc ttgattgttc tttctttttc
     2521 gctattgtaa aattcatgtt atatggaggg ggcaaagttt tcagggtgtt gtttagaatg
     2581 ggaagatgtc ccttgtatca ccatggaccc tcatgataat tttgtttctt tcactttcta
     2641 ctctgttgac aaccattgtc tcctcttatt ttcttttcat tttctgtaac ttttttcgtt
     2701 aaactttagc ttgcatttgt aacgaatttt taaattcact ttcgtttatt tgtcagattg
     2761 taagtacttt ctctaatcac ttttttttca aggcaatcag ggtaattata ttgtacttca
     2821 gcacagtttt agagaacaat tgttataatt aaatgataag gtagaatatt tctgcatata
     2881 aattctggct ggcgtggaaa tattcttatt ggtagaaaca actacatcct ggtaatcatc
     2941 ctgcctttct ctttatggtt acaatgatat acactgtttg agatgaggat aaaatactct
     3001 gagtccaaac cgggcccctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     3061 ctgggcaacg tgctggttgt tgtgctgtct catcattttg gcaaagaatt cactcctcag
     3121 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac
     3181 cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     3241 ctgacttctg ggtaataaag gaaatttatt ttcattgcaa tagtgtgtgg gaattttttg
     3301 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
     3361 ttggtttaga gtttggcaac atatgccata tgctggctgc catgaacaaa ggtggctata
     3421 aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc
     3481 cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc
     3541 cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc
     3601 agtcatagct gtccctcttc tcttatgaac tcgactgcat taatgaatcg gccaacgcgc
     3661 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3721 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3781 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3841 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3901 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3961 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4021 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     4081 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4141 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4201 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4261 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     4321 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4381 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4441 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4501 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4561 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4621 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4681 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4741 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4801 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4861 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4921 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4981 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5041 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5101 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5161 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5221 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5281 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5341 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5401 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5461 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5521 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5581 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     5641 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcactcgag
     5701 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5761 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5821 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     5881 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     5941 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     6001 gcttttttgg aggcctaggc ttttgcaaaa agcttgcatg cctgcaggtc ggccgccacg
     6061 accggtgccg ccaccatccc ctgacccacg cccctgaccc ctcacaagga gacgaccttc
     6121 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt
     6181 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga
     6241 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     6301 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     6361 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     6421 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     6481 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     6541 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     6601 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     6661 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     6721 acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atggctccga
     6781 ccgaagccga cccgggcggc cccgccgacc ccgcacccgc ccccgaggcc caccgactct
     6841 agaggatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     6901 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     6961 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     7021 ttttttcact gc
//
