LOCUS       sequence-407483-        8745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, TCF3_HLF from patient-derived 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8745)
  AUTHORS   Tripathi S, Shirnekhi HK, Gorman SD, Chandra B, Baggett DW, Park CG,
            Somjee R, Lang B, Hosseini SMH, Pioso BJ, Li Y, Iacobucci I, Gao Q, 
            Edmonson MN, Rice SV, Zhou X, Bollinger J, Mitrea DM, White MR, 
            McGrail DJ, Jarosz DF, Yi SS, Babu MM, Mullighan CG, Zhang J, Sahni 
            N, Kriwacki RW
  TITLE     Defining the condensate landscape of fusion oncoproteins.
  JOURNAL   Nat Commun. 2023 Sep 28;14(1):6008. doi: 10.1038/s41467-023-41655-2.
  PUBMED    37770423
REFERENCE   2  (bases 1 to 8745)
  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 
            Commun."; date: "2023-09-28"; volume: "14(1):6008. doi"; pages: " 
            10.1038/s41467-023-41655-2"
COMMENT     Sequence Label: CL20_mEGFP_TCF3_HLF
FEATURES             Location/Qualifiers
     source          1..8745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     220..239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     473..490
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    558..692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(582..601)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     636..655
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(698..893)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      712..847
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    952..973
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(988..993,994..1011,1012..1018)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1026..1042
                     /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     1031..1053
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1050..1066
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1050..1066
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1409..1428
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1427..1482
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1528..1547
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(3506..4216,4217..4219,4220..4222))
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3548..3569)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3897..3916
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4156..4177
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4219..4228
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4304..4507)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4333..4353)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4508..4811
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(5068..5112)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5297..5530
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    5890..6007
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6499..6624
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6671..6928
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6930..7309
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7340..7444
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7445..7513,7514..8305)
                     /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     complement(7663..7682)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8476..8745,1..319)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
       61 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
      121 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      181 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      241 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
      301 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
      361 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
      421 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
      481 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
      541 gattcattaa tgcagcggat ccagacatga taagatacat tgatgagttt ggacaaacca
      601 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
      661 ttgtaaccat tataagctgc aataaacaag ttagcttttt gcaaaagcct aggcctccaa
      721 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
      781 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
      841 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcggatccg
      901 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
      961 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     1021 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     1081 gcttgggctg caggtcgagg gatctccata agagaagagg gacagctatg actgggagta
     1141 gtcaggagag gaggaaaaat ctggctagta aaacatgtaa ggaaaatttt agggatgtta
     1201 aagaaaaaaa taacacaaaa caaaatataa aaaaaatcta acctcaagtc aaggcttttc
     1261 tatggaataa ggaatggaca gcagggggct gtttcatata ctgatgacct ctttatagcc
     1321 aacctttgtt catggcagcc agcatatggg catatgttgc caaactctaa accaaatact
     1381 cattctgatg ttttaaatga tttgccctcc catatgtcct tccgagtgag agacacaaaa
     1441 aattccaaca cactattgca atgaaaataa atttccttta ttagccagaa gtcagatgct
     1501 caaggggctt catgatgtcc ccataatttt tggcagaggg aaaaagatct gctagctcga
     1561 gcagctgaag ctttattgag gcttaagcag tgggttccct agttagccag agagctccca
     1621 ggctcagatc tggtctaacc agagagaccc agtacaggca aaaagcagat cttgtcttct
     1681 ttgggagtga attagccctt ccagtccccc cttttctttt aaaaagtggc taagatctac
     1741 agctgccttg taagtcattg gtcttaaagg taccgtaatc ttatctagaa ccggttcata
     1801 agggtccgtg acgtgcttca tacttagcca ggatattctt gcacttaccc agttctttac
     1861 gcaaatcggc tacctcttga cgaagcgcac tattttcttt ctccaagaaa gatgcacgaa
     1921 tagcaatctg gttctccttc aggcgacgcg catcacggct gcgttttgcc gccatgttgt
     1981 tcttgcggcg gcgcgcccag tacttatcat caaccgtcgg cgtcggaagt cccaaacatg
     2041 ttaatgaggg ggctggttga gagggcaaag ccgcatggtt gtacataagt gatgtactgc
     2101 cagccaatcc atcttcgggg tgagaccccc caaccaggcc ggcatggcgc cctcccaggg
     2161 acattggacc agtgaaccct gatgccagcg ctccgtgccc tggtaacaat gtgtgcatgt
     2221 ctccggccgt gccgacagca tgactacgaa gtacatggat ggcctcatcc agatgatctt
     2281 cgattttgct ctgaagccca tgcagccccc catcgtaaga gggcgataaa gctcccggag
     2341 cgcccgcacg cggccattgg gatgtcccgg ccaacccctg tggagaccct accggagtac
     2401 ttggtgaaga tgagaagtta ttagatgagt ggtctggtga atagatacta gcaagtgcct
     2461 ttccaagtgc gtcaccgctg gaacctgcag tggtaccgcg tgaacccagc aaagaatcag
     2521 cacctgagac agggggtgta tgtgacgaaa ctcccccgta tgtagcgcct ggggcgctgc
     2581 taaacgagct cgcggatggt aaacccccgt ttacctcagc gccgtgtaat tgatacccca
     2641 tgcgttcgtg ttggtgaagc cccccgaagg tcgaactcga cccggagctc ccgactggcc
     2701 cgctacctgg gggcagagga agaggagaag agcccccgcc taacatcggg ccaaatccgg
     2761 cctgtcctgg agggctccac agctctgctg aaggatgcag gcttccatcc gctacgtaga
     2821 aaggggcagg gtaagttgag gaaggagttt tggccgatgg gtaagctgtt gcgtcacggc
     2881 cataatcctc gcctgaagaa ggggggtaga cactactcgg taagccagga ggcactttac
     2941 gcactttctt aggttgagtg tctaaggaac catcagctgc gcgacggcga gaggacccac
     3001 tatagctagg gtagtattgg gacgtcccct tcattcctga agggctaagt ggtccggggc
     3061 tgttcaaagc aagttcacca ctcaggaacc ccgcttgtgt caaaccacct actcccgcat
     3121 cacggccgaa agaggcatac gcgccgcgct caccgctctt tccgcccaaa cccggaccaa
     3181 gaaaagtact agaactcaag gaactgtggc tttcggtgaa gtgagtacct tctgaaaacg
     3241 tgcggcttgg atcgaaggat gaagagcttt gatcaccaga cccccatgat ccgctgcttg
     3301 gacggtcttc aaggcccgac cccccaaact gggcccctgc aagagaggcc gggcgcccct
     3361 tgccattggt tacgggcaaa gggaacatca tagagaaatc aagtaaatca ctaagctcct
     3421 tgtcagtacc aactggggcc atgcgctgag gttggttcat tccccgggaa gcggccgcac
     3481 ttcccccgga gcctccgctt ccgcccttgt acagctcgtc catgccgaga gtgatcccgg
     3541 cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca
     3601 gcttggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg
     3661 gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga
     3721 tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag acgttgtggc
     3781 tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc
     3841 ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct
     3901 tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg
     3961 cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag cactgcacgc
     4021 cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga
     4081 tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga
     4141 acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca
     4201 gctcctcgcc cttgctcacc atggtggcga ccggtggatc ccgggcccgc ggtaccgtcg
     4261 actgcagaat tcggttcact aaacgagccc ggttcactaa acgagctctg cttatataga
     4321 cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt
     4381 ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga
     4441 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc
     4501 gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     4561 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     4621 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     4681 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     4741 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     4801 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
     4861 attaataact agcacgcgtt cgaagcttga gctcggtccc ctcggggttg ggaggtgggt
     4921 ctgaaacgat aatggtgaat atccctgcct aactctattc actatagaaa gtacagcaaa
     4981 aactattctt aaacctacca agcctcctac tatcattatg aataatttta tataccacag
     5041 ccaatttgtt atgttaaacc aattccacaa acttgcccat ttatctaatt ccaataattc
     5101 ttgttcattc ttttcttgct ggttttgcga ttcttcaatt aaggagtgta ttaagcttgt
     5161 gtaattgtta atttctctgt cccactccat ccaggtcgtg tgattccaaa tctgttccag
     5221 agatttatta ctccaactag cattccaagg cacagcagtg gtgcaaatga gttttccaga
     5281 gcaaccccaa atccccagga gctgttgatc ctttaggtat ctttccacag ccaggattct
     5341 tgcctggagc tgcttgatgc cccagactgt gagttgcaac agatgctgtt gcgcctcaat
     5401 agccctcagc aaattgttct gctgctgcac tataccagac aataattgtc tggcctgtac
     5461 cgtcagcgtc attgacgctg cgcccatagt gcttcctgct gctcccaaga acccaaggaa
     5521 caaagctcct attcccactg ctcttttttc tctctgcacc actcttctct ttgccttggt
     5581 gggtgctact cctaatggtt caatttttac tactttatat ttatataatt cacttctcca
     5641 attgtccctc atatctcctc cttatggtgt tttactaaac ttttccatgt tctaatcctc
     5701 atcctgtcta cttgccacac aatcatcacc tgccatctgt tttccataat ccctaatgat
     5761 ctttgctttt cttcttggca ctacttttat gtcactatta tcttgtatta ctactgcccc
     5821 ttcacctttc cagaggagct ttgctggtcc tttccaaagt ggatttctgc tgtccctgta
     5881 ataaacccga aaattttgaa tttttgtaat ttgtttttgt aattctttag tttgtatgtc
     5941 tgttgctatt atgtctacta ttctttcccc tgcactgtac cccccaatcc ccccttttct
     6001 tttaaaattg tggatgaata ctgccatttg tactgctgtc ttaagatgtt cagcctgatc
     6061 tcttacctgt cctataattt tctttaattc tttattcata gattctacta ctccttgact
     6121 ttggggattg tagggaattt ccaggtctga agatcctgct gcttgctgtg cttttttctt
     6181 acttttgttt tgctcttcct ctatcttgtc taaagcttcc ttggtgtctt ttatctctat
     6241 cctttgatgc acacaataga gggttgctac tgtattatat aatgatctaa gttcttctga
     6301 tcctgtctga agggatggtt gtagctgtcc cagtatttgt ctacagcctt ctgatgtttc
     6361 taacaggcca gtattaactg cgaatcgttc tagctccctg cttgcccata ctatatgttt
     6421 taatttatat tttttctttc cccctggcct taaccgaatt ttttcccatc gcgatctaat
     6481 tctcccccgc ttaatactga cgctctcgca cccatctctc tccttctagc ctccgctagt
     6541 caaaattttt ggcgtactca ccagtcgccg cccctcgcct cttgccgtgc gcgcttcagc
     6601 aagccgagtc ctgcgtcgag agagctcctc tggtttccct ttcgctttca agtccctgtt
     6661 cgggcgccac tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta
     6721 ccagagtcac acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga
     6781 ggcttaagca gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac
     6841 cagagagacc cagtacaggc aaaaagcagc tgcttatatg caggatctga gggctcgcca
     6901 ctccccagtc ccgcccaggc cacgcctccc atggtaatag cgatgactaa tacgtagatg
     6961 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
     7021 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
     7081 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
     7141 gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag
     7201 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
     7261 aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtcg acccaggtgg
     7321 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     7381 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     7441 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     7501 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     7561 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     7621 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     7681 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     7741 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     7801 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     7861 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     7921 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     7981 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     8041 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     8101 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     8161 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     8221 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     8281 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     8341 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     8401 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     8461 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     8521 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     8581 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     8641 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     8701 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgg
//
