LOCUS       sequence-182129-        8474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flag tagged mouse Brd4 expression construct for cell transfection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8474)
  AUTHORS   Garcia-Gutierrez P, Mundi M, Garcia-Dominguez M
  TITLE     Association of bromodomain BET proteins with chromatin requires 
            dimerization through the conserved motif B.
  JOURNAL   J Cell Sci. 2012 Aug 1;125(Pt 15):3671-80. Epub 2012 May 17.
  PUBMED    22595521
REFERENCE   2  (bases 1 to 8474)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Sci."; date: "2012-08-1"; volume: "125(Pt 15)"; pages: "3671-80. 
            Epub 2012 May 17"
COMMENT     Sequence Label: RSV-Flag-Brd4
FEATURES             Location/Qualifiers
     source          1..8474
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   34..136
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     224..374
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
     primer_bind     496..512
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     496..512
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        827..1088
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     intron          1130..1226
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     regulatory      1309..1318
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1318..1341
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3571..3588
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5614..5748
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5638..5657)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5692..5711
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5859..5875)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5859..5875)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5872..5894)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5883..5899
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5907..5913,5914..5931,5932..5937))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5952..5973
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6090..6107)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6261..6849)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6341..6360)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7020..7811,7812..7880))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7643..7662
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7881..7985)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8053..8071
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8109..8131)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8444..8466
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(8458..8474,1)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     join(8459..8474,1)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tgaattcaag aattctcatg tttgacagct tatcatcatc aataatatac cttattttgg
       61 attgaagcca atatgataat gagggggtgg agtttgtgac gtggcgcggg gcgtgggaac
      121 ggggcgggtg acgtagtagt gtggcggaag tgtgatgttg caagtgtggc ggaacacatg
      181 taagcgacgg atgtggcaaa agtgacgttt ttggtgtgcg ccggtgtaca caggaagtga
      241 caattttcgc gcggttttag gcggatgttg tagtaaattt gggcgtaacc gagtaagatt
      301 tggccatttt cgcgggaaaa ctgaataaga ggaagtgaaa tctgaataat tttgtgttac
      361 tcatagcgcg taatatttgt ctagggccgc ggggactttg accgtttacg tggagactcg
      421 cccaggtgtt tttctcaggt gttttccgcg ttccgggtca aagttggcgt tttattatta
      481 tagtcagggg gatcttctag aactagtgga tccctatggt gcactctcag tacaatctgc
      541 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
      601 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
      661 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
      721 atctgagggg actagggtgt gtttaggcga aaagcggggc ttcggttgta cgcggttagg
      781 agtcccctca ggatatagta gtttcgcttt tgcataggga gggggaaatg tagtcttatg
      841 caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag
      901 agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag
      961 gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattccgc attgcagaga
     1021 tattgtattt aagtgcctag ctcgatacaa taaacgccat ttgaccattc accacattgg
     1081 tgtgcacctc caagcttctc gaggaactga aaaaccagaa agttaactgg taagtttagt
     1141 ctttttgtct tttatttcag gtcccggatc cggtggtggt gcaaatcaaa gaactgctcc
     1201 tcagtggatg ttgcctttac ttctaggcct gtacggaagt gttacttctg ctctaaaagc
     1261 tgcggaattg tacccgcggc cgcgcgccta ggagctcggt acccccgggc caccatggat
     1321 tacaaggatg acgacgataa gctagccggc acgggcatgt ctacggagag cggccctggg
     1381 acaagattga gaaatctgcc agtaatgggg gatggactag aaacctccca aatgtctaca
     1441 acgcaggccc aggcccaacc ccagccagca aatgcagcca gcaccaatcc tccaccccca
     1501 gagacctcca accctaacaa gcccaagaga cagacaaacc aactgcaata tctgctcaga
     1561 gtggtgctca agacactatg gaaacaccag tttgcgtggc ctttccagca gcccgtggat
     1621 gccgtcaagc tgaacctccc tgattactat aagattatta aaacacccat ggatatggga
     1681 acaataaaga agcgcttgga aaacaactat tactggaatg ctcaggaatg tatccaggac
     1741 ttcaacacta tgtttacaaa ttgttacatc tataacaagc ctggagatga catcgtctta
     1801 atggcagaag ctctggagaa gctcttcttg caaaaaatca atgaactgcc tacagaagaa
     1861 actgagatca tgatagtcca ggcaaaagga agaggacgag ggaggaaaga aacaggggca
     1921 gcaaagcctg gtgtatccac ggtaccaaac acaactcaag catcaacttc tccgcagacc
     1981 cagacgcctc agcagaaccc tcctccacct gtgcaggcca caactcaccc ctttcctgct
     2041 gtcaccccag acctcattgc ccagcctcct gtcatgacaa tggtgccccc tcagccactt
     2101 cagactcctt caccggtacc cccccagcca ccacccccac ctgctccagt tccacagcct
     2161 gtgcagagtc acccgcccat cattgcgacc accccccagc ctgtgaagac aaagaaaggg
     2221 gtgaagagga aagcagatac caccacccct accaccatcg accccattca tgagccaccc
     2281 tcactggccc cagagcccaa gaccgccaag ctgggtcctc ggcgggagag cagcagacct
     2341 gtgaagcctc caaagaagga tgtaccggac tcacagcagc acccagggcc agagaagagc
     2401 agcaagatct ctgagcagct aaagtgctgc agtggcatcc tcaaggagat gtttgccaag
     2461 aaacatgctg cctatgcctg gcctttctac aagcctgtgg atgtggaggc actgggtctg
     2521 cacgactact gtgacatcat caaacatccc atggacatga gcacaatcaa gtctaaacta
     2581 gagtcccgag agtacagaga tgcccaggaa tttggtgctg atgtccgatt gatgttctcc
     2641 aactgctaca agtacaaccc ccctgaccat gaagtggtag ccatggctcg aaaactccag
     2701 gatgtgtttg aaatgcgctt tgccaagatg cctgatgagc ctgaagagcc agttgttaca
     2761 gtgtcctctc ctgcagtgcc accccctaca aaggtggtag ccccaccctc atctagtgac
     2821 agcagcagcg acagttcttc cgacagcgac agttccactg acgactctga ggaagagcga
     2881 gcccagcggc tggctgaact ccaggaacag ctcaaggccg tgcatgagca gcttgcagcc
     2941 ctctcacagc cccagcagaa caaaccaaag aaaaaggaga aggacaagaa ggaaaagaaa
     3001 aaggaaaagc acaaaaagaa agaagaagtg gaggaaaata aaaaaagcaa aaccaaggaa
     3061 cttcctccca aaaagacaaa gaaaaataac agcagcaaca gcaatgtgag caagaaggaa
     3121 ccagtaccca cgaagaccaa gccgcctccc acatatgaat cagaagagga ggataagtgt
     3181 aagcccatgt cttatgagga gaagcggcag ctaagtctag atatcaacaa acttcctggt
     3241 gagaagctag gccgtgtagt acacataatt cagtcaaggg aaccatcact taaaaactcc
     3301 aaccccgatg agattgagat tgactttgag accctgaagc catctacact acgagagttg
     3361 gagcgatatg tcacctcctg tttgcggaag aaaaggaaac ctcaagctga aaaagttgac
     3421 gtgattgctg gttcttccaa gatgaaggga ttctcatcct ctgagtcgga gagcaccagc
     3481 gaatccagct cctctgacag tgaagactct gaaacagaga tggctcccaa gtcaaaaaag
     3541 aaggggcaca ctgggaggga ccagaaaaag catcatcacc atcaccatcc acagatgcag
     3601 ccagccccag ctcctgtgcc ccagcagccg cccccacctc cacagcagcc tcctccaccc
     3661 ccacctccgc agcagcagca gcagcaacct ccacccccac cacctccgcc ctccatgcca
     3721 cagcagactg ccccagcgat gaagtcctcg cccccaccct tcatcactgc ccaggtcccc
     3781 gtcctggaac cacagctgcc aggcagtgtc tttgacccta ttggccactt cacccagccc
     3841 atcttgcacc tgccgcagcc ggagctgcct cctcacctgc cccagccacc tgagcacagc
     3901 actccacccc atctcaacca gcatgctgtg gtctctcctc cagctttgca caatgcgctg
     3961 ccccaacagc catctcggcc cagtaaccga gctgctgctc tgcccccaaa gcctacccga
     4021 cccccagctg tgtcccctgc cctggcccag ccccccctgc tcccacaacc accaatggct
     4081 cagccccccc aagtgctgct ggaggatgaa gagccacctg ccccacccct cacctccatg
     4141 cagatgcagt tgtacctaca gcagctgcag aaggtgcagc ctcccacacc actactccct
     4201 tccgtgaagg tgcagtccca acccccaccg cctttgccgc ctccacccca cccttctgtg
     4261 cagcagcagc agctccagcc acagccaccg ccacccccac ctcctcagcc acagccacca
     4321 ccccagcaac aacaccagcc tccaccacga ccagttcact tgccatccat gcccttttct
     4381 gctcatattc agcagccccc accaccccca ggacagcaac ctactcaccc acccccagga
     4441 cagcagcccc caccaccaca gcctgccaag ccccagcaag tcatccagca tcacccttcc
     4501 ccccggcacc acaagtcaga cccctactca gctggtcatc ttcgtgaggc tccctctccg
     4561 cttatgatac attcccctca gatgccacag ttccagagcc tgacccatca gtctcctccc
     4621 cagcaaaacg tccagcctaa gaagcaggta aagggcaggg ctgagccaca gccaccaggg
     4681 ccagtcatgg gccaaggcca gggatgccca cccgcctcac cggctgccgt gcctatgctg
     4741 tcccaggagc tgcgaccacc ttcagtcgtc cagccccagc ccctggtggt agtaaaggag
     4801 gagaagattc actcaccaat cattcgcagc gagcctttca gcacctcact tcgaccagag
     4861 ccccccaagc acccagagaa catcaaggcc ccagtccacc tgccccagcg gcctgagatg
     4921 aagcctgtag acatagggag gcctgtgatc cggcctccag agcagagtgc acccccacca
     4981 ggggcccctg acaaggacaa acagaagcag gagccaaaaa caccagtggc acccaaaaag
     5041 gacctgaaaa ttaagaacat gggctcctgg gccagcctgg tacagaagca tccgaccacc
     5101 ccatcctcca cagccaagtc ctcaagtgac agctttgagc atttccgccg tgctgctcgg
     5161 gagaaggagg agagggaaaa ggccctgaag gctcaggctg agcatgcaga gaaggagaag
     5221 gagcggctga ggcaggagcg catgaggagc cgagaggatg aagatgcgct ggaacaggca
     5281 cgtcgagccc atgaggaggc acgacggcgc caggagcagc agcagcagca gcagcagcag
     5341 agacaggagc agcagcagca gcagcagcag gcagctgctg tggccgctgc ctctgccccc
     5401 caggcccaga gctcccagcc ccagtctatg ctggaccagc agagggagtt ggcccggaaa
     5461 cgagagcagg agcggaggcg cagggaggca atggcagcta caattgacat gaatttccag
     5521 agtgatcttt tgtcaatatt tgaagaaaat cttttttgag agcacccagg tgacttcgaa
     5581 tgcatagatc tagccggggg atctggccgc ggggatccag acatgataag atacattgat
     5641 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     5701 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     5761 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggttttttc ggatcctcta
     5821 gagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     5881 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     5941 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6001 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     6061 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6121 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6181 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6241 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6301 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6361 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6421 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6481 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6541 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6601 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6661 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6721 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6781 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6841 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6901 tcacgttaag ggattttggt catgtgatta tcaaaaagga tcttcaccta gatcctttta
     6961 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7021 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7081 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7141 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7201 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7261 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7321 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7381 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7441 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7501 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7561 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7621 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7681 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7741 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7801 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7861 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7921 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7981 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     8041 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     8101 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     8161 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     8221 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     8281 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
     8341 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     8401 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     8461 aaaacgacgg ccag
//
