LOCUS       sequence-311527-        6190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses fusion of GAL4 DNA-binding domain and RUNX2Awt IDR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6190)
  AUTHORS   Basu S, Mackowiak SD, Niskanen H, Knezevic D, Asimi V, Grosswendt S,
            Geertsema H, Ali S, Jerkovic I, Ewers H, Mundlos S, Meissner A, 
            Ibrahim DM, Hnisz D
  TITLE     Unblending of Transcriptional Condensates in Human Repeat Expansion 
            Disease.
  JOURNAL   Cell. 2020 May 28;181(5):1062-1079.e30. doi: 
            10.1016/j.cell.2020.04.018. Epub 2020 May 7.
  PUBMED    32386547
REFERENCE   2  (bases 1 to 6190)
  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"; doi: 
            "10.1016/j.cell.2020.04.018"; journalName: "Cell"; date: 
            "2020-05-28- 28"; volume: "181"; issue: "5"; pages: "1062-1079.e30"
COMMENT     Sequence Label: pGAL4-DBD-RUNX2wt-IDR
FEATURES             Location/Qualifiers
     source          1..6190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             710..1150
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     regulatory      1163..1172
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1509..1527)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1510..1527)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1547..1564)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1553..1777
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1823..2251
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1910..1929)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2120..2141
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2260..2280)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2265..2594
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2445..2580
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2507..2526
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2661..3455
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2715..2734)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3325..3344
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3629..3750
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3666..3685)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3720..3739
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3799..3815)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3799..3815)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3812..3834)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3823..3839
                     /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(3847..3853,3854..3871,3872..3877))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3892..3913
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4030..4047)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4201..4789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4281..4300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4960..5751,5752..5820))
                     /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     5583..5602
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5821..5925)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6000..6019)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattca tgaagctact
      721 gtcttctatc gaacaagcat gcgatatttg ccgacttaaa aagctcaagt gctccaaaga
      781 aaaaccgaag tgcgccaagt gtctgaagaa caactgggag tgtcgctact ctcccaaaac
      841 caaaaggtct ccgctgacta gggcacatct gacagaagtg gaatcaaggc tagaaagact
      901 ggaacagcta tttctactga tttttcctcg agaagacctt gacatgattt tgaaaatgga
      961 ttctttacag gatataaaag cattgttaac aggattattt gtacaagata atgtgaataa
     1021 agatgccgtc acagatagat tggcttcagt ggagactgat atgcctctaa cattgagaca
     1081 gcatagaata agtgcgacat catcatcgga agagagtagt aacaaaggtc aaagacagtt
     1141 gactgtatcg gcgatcgcta gtgccaccat ggcatcaaac agcctcttca gcacagtgac
     1201 accatgtcag caaaacttct tttgggatcc gagcaccagc cggcgcttca gccccccctc
     1261 cagcagcctg cagcccggca aaatgagcga cgtgagcccg gtggtggctg cgcaacagca
     1321 gcagcaacag cagcagcagc aacagcagca gcagcagcag caacagcagc agcagcagca
     1381 ggaggcggcg gcggccgctg cggcggcggc ggcggctgcg gcggcggcag ctgcagtgcc
     1441 ccggttacgg ccgccccacg acaaccgcac catggtggag acgtacgctt aagtctagag
     1501 ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct
     1561 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     1621 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     1681 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     1741 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     1801 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     1861 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     1921 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     1981 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2041 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2101 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2161 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2221 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     2281 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2341 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2401 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     2461 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     2521 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     2581 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     2641 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     2701 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     2761 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     2821 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     2881 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     2941 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3001 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3061 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     3121 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     3181 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     3241 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     3301 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     3361 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     3421 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     3481 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     3541 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     3601 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     3661 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3721 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     3781 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3841 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3901 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3961 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4021 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4081 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4141 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4201 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4261 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4321 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4381 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4441 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4501 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4561 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4621 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4681 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4741 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4801 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4861 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4921 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4981 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac ttcccgtcgt
     5041 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5101 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5161 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5221 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5281 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5341 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5401 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5461 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5521 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5581 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5641 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5701 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5761 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5821 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5881 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5941 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     6001 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     6061 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     6121 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     6181 tatacgcgtt
//
