LOCUS       sequence-466517-        5120 bp DNA     circular SYN 25-MAR-2025
DEFINITION  pRSF-DUET1 for protein overexpression of human BRD4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5120)
  AUTHORS   Hsia O, Hinterndorfer M, Cowan AD, Iso K, Ishida T, Sundaramoorthy 
            R, Nakasone MA, Imrichova H, Schatz C, Rukavina A, Husnjak K, Wegner
            M, Correa-Saez A, Craigon C, Casement R, Maniaci C, Testa A, Kaulich
            M, Dikic I, Winter GE, Ciulli A
  TITLE     Targeted protein degradation via intramolecular bivalent glues.
  JOURNAL   Nature. 2024 Mar;627(8002):204-211. doi: 10.1038/s41586-024-07089-6.
            Epub 2024 Feb 21.
  PUBMED    38383787
REFERENCE   2  (bases 1 to 5120)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 25, 2025 from SnapGene Server 7.0.3
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41586-024-07089-6"; journalName: "Nature"; date: 
            "2024-03"; volume: "627"; issue: "8002"; pages: "204-211"
FEATURES             Location/Qualifiers
     source          1..5120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     CDS             97..126
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             136..159
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     promoter        1522..1540
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1541..1565
                     /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)."
     CDS             1674..1718
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1756..1774)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1770..1817
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2047..2862)
                     /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             complement(2047..2862)
                     /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     2770..2789
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2863..2954)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2970..3719)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(2980..2997)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3220..3239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3915..4997)
                     /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     4959..4978
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4998..5075)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg ggcagccatc accatcatca tcaccatcac
      121 caccacggtg gttctctgga agttctgttc caggggcccg gatccaccaa ccccccgccc
      181 ccagagacct ccaaccctaa caagcccaag aggcagacca accaactgca atacctgctc
      241 agagtggtgc tcaagacact atggaaacac cagtttgcat ggcctttcca gcagcctgtg
      301 gatgccgtca agctgaacct ccctgattac tataagatca ttaaaacgcc tatggatatg
      361 ggaacaataa agaagcgctt ggaaaacaac tattactgga atgctcagga atgtatccag
      421 gacttcaaca ctatgtttac aaattgttac atctacaaca agcctggaga tgacatagtc
      481 ttaatggcag aagctctgga aaagctcttc ttgcaaaaaa taaatgagct acccacagaa
      541 gaaaccgaga tcatgatagt ccaggcaaaa ggaagaggac gtgggaggaa agaaacaggg
      601 acagcaaaac ctggcgtttc cacggtacca aacacaactc aagcatcgac tcctccgcag
      661 acccagaccc ctcagccgaa tcctcctcct gtgcaggcca cgcctcaccc cttccctgcc
      721 gtcaccccgg acctcatcgt ccagacccct gtcatgacag tggtgcctcc ccagccactg
      781 cagacgcccc cgccagtgcc cccccagcca caacccccac ccgctccagc tccccagccc
      841 gtacagagcc acccacccat catcgcggcc accccacagc ctgtgaagac aaagaaggga
      901 gtgaagagga aagcagacac caccaccccc accaccattg accccattca cgagccaccc
      961 tcgctgcccc cggagcccaa gaccaccaag ctgggccagc ggcgggagag cagccggcct
     1021 gtgaaacctc caaagaagga cgtgcccgac tctcagcagc acccagcacc agagaagagc
     1081 agcaaggtct cggagcagct caagtgctgc agcggcatcc tcaaggagat gtttgccaag
     1141 aagcacgccg cctacgcctg gcccttctac aagcctgtgg acgtggaggc actgggccta
     1201 cacgactact gtgacatcat caagcacccc atggacatga gcacaatcaa gtctaaactg
     1261 gaggcccgtg agtaccgtga tgctcaggag tttggtgctg acgtccgatt gatgttctcc
     1321 aactgctata agtacaaccc tcctgaccat gaggtggtgg ccatggcccg caagctccag
     1381 gatgtgttcg aaatgcgctt tgccaagatg ccggactaag aattcgagct cggcgcgcct
     1441 gcaggtcgac aagcttgcgg ccgcataatg cttaagtcga acagaaagta atcgtattgt
     1501 acacggccgc ataatcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca
     1561 attccccatc ttagtatatt agttaagtat aagaaggaga tatacatatg gcagatctca
     1621 attggatatc ggccggccac gcgatcgctg acgtcggtac cctcgagtct ggtaaagaaa
     1681 ccgctgctgc gaaatttgaa cgccagcaca tggactcgtc tactagcgca gcttaattaa
     1741 cctaggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     1801 tcttgagggg ttttttgctg aaacctcagg catttgagaa gcacacggtc acactgcttc
     1861 cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc
     1921 ctgaaccgac gacaagctga cgaccgggtc tccgcaagtg gcacttttcg gggaaatgtg
     1981 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat
     2041 taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
     2101 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     2161 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     2221 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     2281 gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac
     2341 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     2401 tgattgcgcc tgagcgagac gaaatacgcg gtcgctgtta aaaggacaat tacaaacagg
     2461 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     2521 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     2581 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     2641 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     2701 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     2761 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     2821 tcgcggccta gagcaagacg tttcccgttg aatatggctc atactcttcc tttttcaata
     2881 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     2941 gaaaaataaa caaataggca tgcagcgctc ttccgcttcc tcgctcactg actcgctacg
     3001 ctcggtcgtt cgactgcggc gagcggtgtc agctcactca aaagcggtaa tacggttatc
     3061 cacagaatca ggggataaag ccggaaagaa catgtgagca aaaagcaaag caccggaaga
     3121 agccaacgcc gcaggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3181 gacgctcaag ccagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3241 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3301 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgttgg tatctcagtt
     3361 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3421 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3481 cactggcagc agccattggt aactgattta gaggactttg tcttgaagtt atgcacctgt
     3541 taaggctaaa ctgaaagaac agattttggt gagtgcggtc ctccaaccca cttaccttgg
     3601 ttcaaagagt tggtagctca gcgaaccttg agaaaaccac cgttggtagc ggtggttttt
     3661 ctttatttat gagatgatga atcaatcggt ctatcaagtc aacgaacagc tattccgtta
     3721 ctctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag ccccatacga
     3781 tataagttgt aattctcatg ttagtcatgc cccgcgccca ccggaaggag ctgactgggt
     3841 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     3901 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3961 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     4021 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     4081 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     4141 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc
     4201 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
     4261 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
     4321 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
     4381 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
     4441 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
     4501 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
     4561 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
     4621 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
     4681 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
     4741 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
     4801 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     4861 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     4921 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     4981 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     5041 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     5101 cgactcctgc attaggaaat
//
