LOCUS       V018467                 9538 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018467
VERSION     V018467
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             932..5017
                     /label="BRD4(NM_058243)"
                     /note="BRD4(NM_058243)"
                     /gene="BRD4"
     CDS             5039..5104
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    5133..5357
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5403..5831
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5845..6174
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6241..7035
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    7211..7332
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7381..7397)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7405..7421
                     /bound_moiety="lac repressor encoded by lacI"
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(7429..7459)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7474..7495
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7783..8371)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8542..9402)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(9403..9507)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatccgccac catgtctgcg gagagcggcc ctgggacgag
      961 attgagaaat ctgccagtaa tgggggatgg actagaaact tcccaaatgt ctacaacaca
     1021 ggcccaggcc caaccccagc cagccaacgc agccagcacc aaccccccgc ccccagagac
     1081 ctccaaccct aacaagccca agaggcagac caaccaactg caatacctgc tcagagtggt
     1141 gctcaagaca ctatggaaac accagtttgc atggcctttc cagcagcctg tggatgccgt
     1201 caagctgaac ctccctgatt actataagat cattaaaacg cctatggata tgggaacaat
     1261 aaagaagcgc ttggaaaaca actattactg gaatgctcag gaatgtatcc aggacttcaa
     1321 cactatgttt acaaattgtt acatctacaa caagcctgga gatgacatag tcttaatggc
     1381 agaagctctg gaaaagctct tcttgcaaaa aataaatgag ctacccacag aagaaaccga
     1441 gatcatgata gtccaggcaa aaggaagagg acgtgggagg aaagaaacag ggacagcaaa
     1501 acctggcgtt tccacggtac caaacacaac tcaagcatcg actcctccgc agacccagac
     1561 ccctcagccg aatcctcctc ctgtgcaggc cacgcctcac cccttccctg ccgtcacccc
     1621 ggacctcatc gtccagaccc ctgtcatgac agtggtgcct ccccagccac tgcagacgcc
     1681 cccgccagtg cccccccagc cacaaccccc acccgctcca gctccccagc ccgtacagag
     1741 ccacccaccc atcatcgcgg ccaccccaca gcctgtgaag acaaagaagg gagtgaagag
     1801 gaaagcagac accaccaccc ccaccaccat tgaccccatt cacgagccac cctcgctgcc
     1861 cccggagccc aagaccacca agctgggcca gcggcgggag agcagccggc ccgtgaaacc
     1921 tccaaagaag gacgtgcccg actctcagca gcacccagca ccagagaaga gcagcaaggt
     1981 ctcggagcag ctcaagtgct gcagcggcat cctcaaggag atgtttgcca agaagcacgc
     2041 cgcctacgcc tggcccttct acaagcctgt ggacgtggag gcactgggcc tacacgacta
     2101 ctgtgacatc atcaagcacc ccatggacat gagcacaatc aagtctaaac tggaggcccg
     2161 tgagtaccgt gatgctcagg agtttggtgc tgacgtccga ttgatgttct ccaactgcta
     2221 taagtacaac cctcctgacc atgaggtggt ggccatggcc cgcaagctcc aggatgtgtt
     2281 cgaaatgcgc tttgccaaga tgccggacga gcctgaggag ccagtggtgg ccgtgtcctc
     2341 cccggcagtg ccccctccca ccaaggttgt ggccccgccc tcatccagcg acagcagcag
     2401 cgatagctcc tcggacagtg acagttcgac tgatgactct gaggaggagc gagcccagcg
     2461 gctggctgag ctccaggagc agctcaaagc cgtgcacgag cagcttgcag ccctctctca
     2521 gccccagcag aacaaaccaa agaaaaagga gaaagacaag aaggaaaaga aaaaagaaaa
     2581 gcacaaaagg aaagaggaag tggaagagaa taaaaaaagc aaagccaagg aacctcctcc
     2641 taaaaagacg aagaaaaata atagcagcaa cagcaatgtg agcaagaagg agccagcgcc
     2701 catgaagagc aagccccctc ccacgtatga gtcggaggaa gaggacaagt gcaagcctat
     2761 gtcctatgag gagaagcggc agctcagctt ggacatcaac aagctccccg gcgagaagct
     2821 gggccgcgtg gtgcacatca tccagtcacg ggagccctcc ctgaagaatt ccaaccccga
     2881 cgagattgaa atcgactttg agaccctgaa gccgtccaca ctgcgtgagc tggagcgcta
     2941 tgtcacctcc tgtttgcgga agaaaaggaa acctcaagct gagaaagttg atgtgattgc
     3001 cggctcctcc aagatgaagg gcttctcgtc ctcagagtcg gagagctcca gtgagtccag
     3061 ctcctctgac agcgaagact ccgaaacaga gatggctccg aagtcaaaaa agaaggggca
     3121 ccccgggagg gagcagaaga agcaccatca tcaccaccat cagcagatgc agcaggcccc
     3181 ggctcctgtg ccccagcagc cgcccccgcc tccccagcag cccccaccgc ctccacctcc
     3241 gcagcagcaa cagcagccgc cacccccgcc tcccccaccc tccatgccgc agcaggcagc
     3301 cccggcgatg aagtcctcgc ccccaccctt cattgccacc caggtgcccg tcctggagcc
     3361 ccagctccca ggcagcgtct ttgaccccat cggccacttc acccagccca tcctgcacct
     3421 gccgcagcct gagctgcccc ctcacctgcc ccagccgcct gagcacagca ctccacccca
     3481 tctcaaccag cacgcagtgg tctctcctcc agctttgcac aacgcactac cccagcagcc
     3541 atcacggccc agcaaccgag ccgctgccct gcctcccaag cccgcccggc ccccagccgt
     3601 gtcaccagcc ttgacccaaa cacccctgct cccacagccc cccatggccc aaccccccca
     3661 agtgctgctg gaggatgaag agccacctgc cccacccctc acctccatgc agatgcagct
     3721 gtacctgcag cagctgcaga aggtgcagcc ccctacgccg ctactccctt ccgtgaaggt
     3781 gcagtcccag cccccacccc ccctgccgcc cccaccccac ccctctgtgc agcagcagct
     3841 gcagcagcag ccgccaccac ccccaccacc ccagccccag cctccacccc agcagcagca
     3901 tcagccccct ccacggcccg tgcacttgca gcccatgcag ttttccaccc acatccaaca
     3961 gcccccgcca ccccagggcc agcagccccc ccatccgccc ccaggccagc agccaccccc
     4021 gccgcagcct gccaagcctc agcaagtcat ccagcaccac cattcacccc ggcaccacaa
     4081 gtcggacccc tactcaaccg gtcacctccg cgaagccccc tccccgctta tgatacattc
     4141 cccccagatg tcacagttcc agagcctgac ccaccagtct ccaccccagc aaaacgtcca
     4201 gcctaagaaa caggagctgc gtgctgcctc cgtggtccag ccccagcccc ttgtggtggt
     4261 gaaggaggag aagatccact cacccatcat ccgcagcgag cccttcagcc cctcgctgcg
     4321 gccggagccc cccaagcacc cggagagcat caaggccccc gtccacctgc cccagcggcc
     4381 ggaaatgaag cctgtggatg tcgggaggcc tgtgatccgg cccccagagc agaacgcacc
     4441 gccaccaggg gcccctgaca aggacaaaca gaaacaggag ccgaagactc cagttgcgcc
     4501 caaaaaggac ctgaaaatca agaacatggg ctcctgggcc agcctagtgc agaagcatcc
     4561 gaccaccccc tcctccacag ccaagtcatc cagcgacagc ttcgagcagt tccgccgcgc
     4621 cgctcgggag aaagaggagc gtgagaaggc cctgaaggct caggccgagc acgctgagaa
     4681 ggagaaggag cggctgcggc aggagcgcat gaggagccga gaggacgagg atgcgctgga
     4741 gcaggcccgg cgggcccatg aggaggcacg tcggcgccag gagcagcagc agcagcagcg
     4801 ccaggagcaa cagcagcagc agcaacagca agcagctgcg gtggctgccg ccgccacccc
     4861 acaggcccag agctcccagc cccagtccat gctggaccag cagagggagt tggcccggaa
     4921 gcgggagcag gagcgaagac gccgggaagc catggcagct accattgaca tgaatttcca
     4981 gagtgatcta ttgtcaatat ttgaagaaaa tcttttcctc gagtctagag ggcccttcga
     5041 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga
     5101 caagtgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     5161 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     5221 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     5281 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     5341 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     5401 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     5461 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5521 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     5581 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5641 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5701 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5761 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5821 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     5881 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     5941 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     6001 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     6061 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     6121 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     6181 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     6241 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     6301 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     6361 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     6421 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     6481 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     6541 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     6601 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     6661 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     6721 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     6781 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6841 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6901 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     6961 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     7021 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa
     7081 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     7141 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     7201 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     7261 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     7321 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     7381 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     7441 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     7501 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     7561 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     7621 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     7681 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7741 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7801 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7861 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7921 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7981 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     8041 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     8101 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     8161 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     8221 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     8281 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     8341 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     8401 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     8461 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     8521 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     8581 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     8641 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     8701 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8761 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8821 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8881 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8941 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     9001 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     9061 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     9121 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     9181 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     9241 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     9301 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     9361 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     9421 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     9481 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
