LOCUS       sequence-387505-        5689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of BRD4 Bromodomain (BD1) in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5689)
  AUTHORS   Kim JH, Pandit N, Yoo M, Park TH, Choi JU, Park CH, Jung KY, Lee BI
  TITLE     Crystal structure of [1,2,4]triazolo[4,3-b]pyridazine derivatives as
            BRD4 bromodomain inhibitors and structure-activity relationship 
            study.
  JOURNAL   Sci Rep. 2023 Jul 4;13(1):10805. doi: 10.1038/s41598-023-37527-w.
  PUBMED    37402749
REFERENCE   2  (bases 1 to 5689)
  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: "Sci Rep."; 
            date: "2023-07-4"; volume: "13(1):10805. doi"; pages: " 
            10.1038/s41598-023-37527-w"
COMMENT     Sequence Label: pHis-BRD4 BD1
FEATURES             Location/Qualifiers
     source          1..5689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1287..1478
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1472..1494)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1580..1719
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1635..1654
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1734..1751)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1905..2493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1985..2004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2604..2623)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2615..3430
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2615..3430
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2688..2707)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(3522..3977)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3659..3680)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3871..3890
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(4014..4061)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4057..4075
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4138..4161)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             complement(4549..4569)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(4591..4608)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4651..4675
                     /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)."
     primer_bind     complement(4675..4694)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4676..4694)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4782..4801)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5007..5084
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5085..5689,1..478)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5104..5123)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
       61 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
      121 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
      181 gcgcgttggt gcggacatct cggtagtggg atacgacgat accgaagaca gctcatgtta
      241 tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
      301 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
      361 actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt
      421 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
      481 gcaacgcaat taatgtaagt tagctcactc attaggcacc gggatctcga ccgatgccct
      541 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg
      601 cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg
      661 tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg
      721 tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt
      781 tcggcgagaa gcaggccatt atcgccggca tggcggcccc acgggtgcgc atgatcgtgc
      841 tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct tactggttag cagaatgaat
      901 caccgatacg cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa
      961 caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc
     1021 cctgcaccat tatgttccgg atctgcatcg caggatgctg ctggctaccc tgtggaacac
     1081 ctacatctgt attaacgaag cgctggcatt gaccctgagt gatttttctc tggtcccgcc
     1141 gcatccatac cgccagttgt ttaccctcac aacgttccag taaccgggca tgttcatcat
     1201 cagtaacccg tatcgtgagc atcctctctc gtttcatcgg tatcattacc cccatgaaca
     1261 gaaatccccc ttacacggag gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg
     1321 cccgctttat cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg
     1381 atgaacaggc agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct
     1441 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     1501 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     1561 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata
     1621 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccatt gcggtgtgaa
     1681 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
     1741 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1801 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1861 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1921 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1981 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2041 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2101 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2161 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2221 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2281 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2341 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2401 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2461 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2521 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgaa caataaaact
     2581 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     2641 ttgctctagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2701 tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc
     2761 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     2821 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     2881 tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt
     2941 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     3001 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     3061 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     3121 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     3181 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     3241 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     3301 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3361 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3421 gtttttctaa gaattaattc atgagcggat acatatttga atgtatttag aaaaataaac
     3481 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     3541 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3601 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3661 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3721 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     3781 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3841 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     3901 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3961 gccgctacag ggcgcgtccc attcgccaat ccggatatag ttcctccttt cagcaaaaaa
     4021 cccctcaaga cccgtttaga ggccccaagg ggttatgcta gttattgctc agcggtggca
     4081 gcagccaact cagcttcctt tcgggctttg tttagcagcc taggtattaa tcaattagtg
     4141 gtggtggtgg tggtggtggt gctcgagtta ttcttctgtg ggtagctcat ttattttttg
     4201 caagaagagc ttttccagag cttctgccat taagactatg tcatctccag gcttgttgta
     4261 gatgtaacaa tttgtaaaca tagtgttgaa gtcctggata cattcctgag cattccagta
     4321 atagttgttt tccaagcgct tctttattgt tcccatatcc ataggcgttt taatgatctt
     4381 atagtaatca gggaggttca gcttgacggc atccacaggc tgctggaaag gccatgcaaa
     4441 ctggtgtttc catagtgtct tgagcaccac tctgagcagg tattgcagtt ggttggtctg
     4501 cctcttgggc ttgttagggt tggaggtctc tgggggcggg gggttggatc cctgaaaata
     4561 caggttttca gatctcggaa ccagcgatcc atggtgatgg tggtgatgca tatgtatatc
     4621 tccttcttaa agttaattta aatctagagg ggaattgtta tccgctcaca attcccctat
     4681 agtgagtcgt attaatttcg cgggatcgag atcgatctcg atcctctacg ccggacgcat
     4741 cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac
     4801 cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat
     4861 ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct
     4921 tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc
     4981 gcataaggga gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc tttcgcggta
     5041 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
     5101 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     5161 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     5221 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     5281 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     5341 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     5401 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     5461 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     5521 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
     5581 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     5641 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggca
//
