LOCUS       sequence-426059-        6053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses BCL6 BTB(1-129)-mCherry-FUS (IDR)-FRB in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6053)
  AUTHORS   Hernandez-Candia CN, Brady BR, Harrison E, Tucker CL
  TITLE     A platform to induce and mature biomolecular condensates using 
            chemicals and light.
  JOURNAL   Nat Chem Biol. 2024 Jan 8. doi: 10.1038/s41589-023-01520-1.
  PUBMED    38191942
REFERENCE   2  (bases 1 to 6053)
  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: "Nat Chem 
            Biol. 2024 Jan 8. doi: 10.1038/s41589-023-01520-1."
COMMENT     Sequence Label: B2088_BTB-IDR-FRB
FEATURES             Location/Qualifiers
     source          1..6053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      597..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1056..1065
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1062..1769
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1207..1225)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1475..1494
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2165..2176
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             2439..2717
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     polyA_signal    2841..2962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2878..2897)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2932..2951
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2969..3424)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3106..3127)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3318..3337
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3451..3555
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3557..3914
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3557..3748
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3580..3600)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3765..3900
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3949..4743
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3949..4743
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4003..4022)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4613..4632
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4931..4950)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4975..5022
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5351..5939
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5840..5859
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcca
      601 ccatggcctc gccggctgac agctgtatcc agttcacccg ccatgccagt gatgttcttc
      661 tcaaccttaa tcgtctccgg agtcgagaca tcttgactga tgttgtcatt gttgtgagcc
      721 gtgagcagtt tagagcccat aaaacggtcc tcatggcctg cagtggcctg ttctatagca
      781 tctttacaga ccagttgaaa tgcaacctta gtgtgatcaa tctagatcct gagatcaacc
      841 ctgagggatt ctgcatcctc ctggacttca tgtacacatc tcggctcaat ttgcgggagg
      901 gcaacatcat ggctgtgatg gccacggcta tgtacctgca gatggagcat gttgtggaca
      961 cttgccggaa gtttattaag gccagtgaag gcggaagcgg cggaaagctt cgaattctgc
     1021 agtcgacggt accgcgggcc cgggatccac cggtcgccac catggtgagc aagggcgagg
     1081 aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg
     1141 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
     1201 agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt
     1261 cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact
     1321 acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     1381 gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg
     1441 tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg
     1501 gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca
     1561 agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca
     1621 aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca
     1681 cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact
     1741 ccaccggcgg catggacgag ctgtacaaag gtggtagtct cgagatggcc tcaaacgatt
     1801 atacccaaca agcaacccaa agctatgggg cctaccccac ccagcccggg cagggctatt
     1861 cccagcagag cagtcagccc tacggacagc agagttacag tggttatagc cagtccacgg
     1921 acacttcagg ctatggccag agcagctatt cttcttatgg ccagagccag aacacaggct
     1981 atggaactca gtcaactccc cagggatatg gctcgactgg cggctatggt agtagccaga
     2041 gctcccaatc gtcttacggg cagcagtcct cctatcctgg ctatggccag cagccagctc
     2101 ccagcagcac ctcgggaagt tacggtagca gttctcagag cagcagctat gggcagcccc
     2161 agagtgggag ctacagccag cagcctagct atggtggaca gcagcaaagc tatggacagc
     2221 agcaaagcta taatccccct cagggctatg gacagcagaa ccagtacaac agcagcagtg
     2281 gtggtggagg tggaggtgga ggtggaggta actatggcca agatcaatcc tccatgagta
     2341 gtggtggtgg cagtggtggc ggttatggca atcaagacca gagtggtgga ggtggcagcg
     2401 gtggctatgg acagcaggac cgtggaggag caagtggaat cctctggcat gagatgtggc
     2461 atgaaggcct ggaagaggca tctcgtttgt actttgggga aaggaacgtg aaaggcatgt
     2521 ttgaggtgct ggagcccttg catgctatga tggaacgggg cccccagact ctgaaggaaa
     2581 catcctttaa tcaggcctat ggtcgagatt taatggaggc ccaagagtgg tgcaggaagt
     2641 acatgaaatc agggaatgtc aaggacctcc tccaagcctg ggacctctat tatcatgtgt
     2701 tccgacgaat ctcaaagtag cggccgcgac tctagatcat aatcagccat accacatttg
     2761 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     2821 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     2881 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     2941 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc
     3001 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     3061 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     3121 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     3181 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     3241 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     3301 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     3361 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     3421 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3481 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3541 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     3601 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3661 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3721 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3781 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3841 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3901 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3961 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4021 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4081 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     4141 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4201 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4261 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4321 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4381 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4441 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     4501 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4561 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4621 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4681 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4741 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4801 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4861 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     4921 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     4981 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     5041 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     5101 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     5161 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     5221 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5281 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5341 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5401 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5461 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5521 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5581 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5641 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5701 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5761 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5821 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5881 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5941 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6001 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
