LOCUS       V009183                 6831 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009183
VERSION     V009183
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6831)
  AUTHORS   Kulman JD.
  TITLE     Mammalian expression vector for doxycycline-regulated production of
            enzymatically biotinylated proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6831)
  AUTHORS   Kulman JD.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-JUN-2014) Puget Sound Blood Center Research Institute,
            Puget Sound Blood Center, 1551 Eastlake Ave E #100, Seattle, WA
            98102, USA
REFERENCE   3  (bases 1 to 6831)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6831)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (21-JUN-2014) Puget Sound Blood Center Research Institute, Puget
            Sound Blood Center, 1551 Eastlake Ave E #100, Seattle, WA 98102,
            USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(29..991)
                     /label="BirA"
                     /note="E. coli biotin protein ligase"
     CDS             complement(1004..1027)
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     regulatory      1027..1036
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1175..1445
                     /label="tetracycline response element"
                     /note="contains seven copies of the tetracycline operator
                     tetO"
     promoter        1478..1515
                     /label="minimal CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     misc_feature    1654..1686
                     /label="multiple cloning site"
                     /note="multiple cloning site"
     CDS             1693..1734
                     /codon_start=1
                     /product="biotin acceptor peptide"
                     /label="biotin acceptor peptide"
                     /note="contains lysine residue to which d-biotin is
                     enzymatically conjugated by an isopeptide linkage"
                     /protein_id="AIR07421.1"
                     /translation="MNDIFEAQKIEWH"
     misc_feature    1764..2350
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             2405..3121
                     /label="EGFP"
                     /note="enhanced GFP"
     intron          3155..3727
                     /label="beta-globin intron"
                     /note="intron from rabbit beta-globin gene"
     CDS             3731..3853
                     /gene="HBB"
                     /label="Hemoglobin subunit beta"
                     /note="Hemoglobin subunit beta from Colobus guereza.
                     Accession#: Q9TT33"
     polyA_signal    3924..3979
                     /label="beta-globin poly(A) signal"
                     /note="rabbit beta-globin polyadenylation signal (Gil and
                     Proudfoot, 1987)"
     rep_origin      complement(4541..5129)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5303..6160)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6161..6265)
                     /label="AmpR promoter"
     polyA_signal    6696..6830
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tctagaacta ggtcgacgcg gccgcttatt tttctgcact acgcagggat atttcaccgc
       61 ccatccaggg ttttattatt ccatcctgct caagtaataa agccccctgt ttgtctattc
      121 cgcgtgaaat gccaaatatt tctttatcac caatgataag tttcactggg cgattaataa
      181 aattatccag cttttcccag cgcgacagat aaggtgccaa tccttcttgt tcgaagagtt
      241 ccaacgcagc acgtaattca cgtattagca tggccgccaa cgtattacga tcgagattga
      301 tccccgcttc ctgcagcgtg atccacccct gattaacgac actctcttca acacggcgca
      361 ttgccatgtt gatcccggct ccaatgacta tttgcgccgc atcgccagtt ttgccagtca
      421 gctccaccag aatgcctgcc agcttgcgat cctgcagata gaggtcatta ggccatttaa
      481 cacgaacttt atctgcaccc agcttgcgta atacttccgc catcacgata ccgataacca
      541 gacttaaacc aatcgccgcc gccgggcctt gttccagacg ccagaacatc gacaaatata
      601 agtttgcgcc aaaaggcgaa aaccatttcc gaccccggcg accacggcca gcctgctggt
      661 attctgcaat gcaagcatcg cccgatttaa gctctccgat acgatcaaga aggtactgat
      721 tcgtggagtc aatcactggc agcacggcta cactaccgcc atccagctga cccaatatct
      781 gtttagcatt aagtaactgg ataggctcag gcaggctgta tcctttaccc ggaacggtaa
      841 agacatcaac gccccagtca cgcagtgtct gaatgtgttt attaatagcc gcccggctca
      901 ttcccagcgt ttcacccaac tgctcgccag agtgaaattc accgttcgct aacagggcaa
      961 tcaatttcag tggcacggtg ttatccttca tggatctcgg gctcttatcg tcgtcatcct
     1021 tgtaatccat ggtggcggca tgcaggaatt cggggccgcg gaggctggat cggtcccggt
     1081 gtcttctatg gaggtcaaaa cagcgtggat ggcgtctcca ggcgatctga cggttcacta
     1141 aacgagctct gcttatatag gtcgagttta ccactcccta tcagtgatag agaaaagtga
     1201 aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc
     1261 cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa
     1321 aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta
     1381 ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat
     1441 agagaaaagt gaaagtcgag ctcggtaccc gggtcgagta ggcgtgtacg gtgggaggcc
     1501 tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt
     1561 tttgacctcc atagaagaca ccgggaccga tccagcctcc gcggccccga attcgagctc
     1621 ggtacccggg gatcctctag tcagctgacg cgtgctagca ctagtgatat cgggccggcc
     1681 ctcgagggcc gcttgaacga catttttgaa gcgcaaaaaa ttgaatggca ttgacgagca
     1741 tgcatctagg gcggccaatt ccgcccctct ccctcccccc cccctaacgt tactggccga
     1801 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt gattttccac catattgccg
     1861 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     1921 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     1981 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     2041 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     2101 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     2161 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     2221 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
     2281 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataagct
     2341 tgccacaacc cgggatcctc tagagtcgac ggtaccgcgg gcccgggatc caccggtcgc
     2401 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2461 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     2521 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2581 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     2641 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2701 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2761 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2821 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     2881 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     2941 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     3001 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     3061 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     3121 gtaaagcggc catctctaga ctgagaactt cagggtgagt ttggggaccc ttgattgttc
     3181 tttctttttc gctattgaaa aattcatgtt atatggaggg ggcaaagttt tcagggtgtt
     3241 gtttagaatg ggaagatgtc ccttgtatca ccatggaccc tcatgataat tttgtttctt
     3301 tcactttcta ctctgttgac aaccattgtc tcctcttatt ttcttttcat tttctgtaac
     3361 ttttttcgtt aaactttagc ttgcatttgt aacgaatttt taaattcact ttcgtttatt
     3421 tgtcagattg taagtacttt ctctaatcac ttttttttca aggcaatcag ggtaattata
     3481 ttgtacttca gcacagtttt agagaacaat tgttataatt aaatgataag gtagaatatt
     3541 tctgcatata aattctggct ggcgtggaaa tattcttatt ggtagaaaca actacatcct
     3601 ggtaatcatc ctgcctttct ctttatggtt acaatgatat acactgtttg agatgaggat
     3661 aaaatactct gagtccaaac cgggcccctc tgctaaccat gttcatgcct tcttcttttt
     3721 cctacagctc ctgggcaacg tgctggttgt tgtgctgtct catcattttg gcaaagaatt
     3781 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     3841 tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc
     3901 ccttgagcat ctgacttctg ggtaataaag gaaatttatt ttcattgcaa tagtgtgtgg
     3961 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca
     4021 gaatgagtat ttggtttaga gtttggcaac atatgccata tgctggctgc catgaacaaa
     4081 ggtggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     4141 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     4201 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     4261 gactactccc agtcatagct gtccctcttc tcttatgaac tcgactgcat taatgaatcg
     4321 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4381 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4441 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4501 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4561 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4621 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4681 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     4741 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4801 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4861 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4921 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4981 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5041 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5101 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5161 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5221 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5281 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5341 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5401 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5461 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5521 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5581 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5641 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5701 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5761 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5821 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5881 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5941 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6001 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6061 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6121 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6181 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6241 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     6301 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     6361 ttcactcgac gctggtcgag ctgatacttc ccgtccgcca ggggacatgc cggcgatgct
     6421 gaaggtcgcg cgcattcccg atgaagaggc cggttaccgc ctgttgacct ggtgggacgg
     6481 gcagggcgcc gcccgagtct tgcctcggcg gcgggcgctc tgctcatgga gcgcgcgtcc
     6541 ggggaccttg cacagatagc gtggtccggc cagacgacga ggcttgcagg atcataatca
     6601 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     6661 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     6721 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6781 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc c
//
