LOCUS       sequence-408113-        8178 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, LEF1_TAF8 from patient-derived 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8178)
  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 8178)
  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_LEF1_TAF8
FEATURES             Location/Qualifiers
     source          1..8178
                     /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(2939..3649,3650..3652,3653..3655))
                     /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(2981..3002)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3330..3349
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3589..3610
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3652..3661
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3737..3940)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3766..3786)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3941..4244
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4501..4545)
                     /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    4730..4963
                     /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    5323..5440
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5932..6057
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6104..6361
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6363..6742
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6773..6877
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6878..6946,6947..7738)
                     /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(7096..7115)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7909..8178,1..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 ccggttcacg
     1801 aaagtgactt cttacggcga attttaggct tctttacagg gcgtaaataa ggattatcaa
     1861 tgatgttttc ctcgccgtta cgactacccg acaaggaagg attctgctga agcacgctgg
     1921 tgttttcttt ctcagcgcca ctatcctcca tagaaatgtg aagagccaga ttctctgtgt
     1981 ccgtttgctc gtcctgctcg ctggaatcgg tctcctccat ttgctgcatt tccaattcgc
     2041 tgggcagcag cgcggtaagg taagggatag taaatgggcg tgctgcaata aggggaaagg
     2101 tcgaaacatc gtccttgaac agcgactgag tctccccggt ttttgccatg aaacgtgtca
     2161 acgcgcgttc gacatcgcga cgttgggacg ctgctttttc gcgtaaaacc tgatagtcgg
     2221 agacgggctc gcgatacgtg ggggtcttga tgtaagtatg ggggtcaggg aactcgggaa
     2281 agtgagaagg gatgtgaggc ggatgcgggc gattttggcc cgcagtcaat gctttcgggg
     2341 tgactggttg gttggtcact ggtggggccg tgatgaccat acgctggcta cgtttcgcat
     2401 aggcaggaag cgtgtctaca ttgaaaccca tctccaccag agttactacg atgtccgaca
     2461 aagttggttg tgtgcgggcc gtgtgctcac agtaagattt ggcagagcgt ccgatttctg
     2521 agatatatga ctgtaacatt tcggtcaagg tttcaacgct cgccttttca gctgactcga
     2581 agcctgcttc tgtcagtaag ctcgatacga ctacctgtaa cgtacgacga cgtgctaaat
     2641 gataattgtc cgcgggattg gttgattgct tggatccaga gcgtgtacgg ggaattgggg
     2701 gtgataagga gccgttgctc atatatggat cgttattcat attaggcatc ataatatacc
     2761 ccgagtaact ggagtagctc ggtcccttat tgtacagacc accatctggg tgtttaccat
     2821 cgtcagggtg ttcacgtgcc ttgtcgtgat atggctcctg gctggtttga gcttggcgtg
     2881 ccacctcggc cattccccgg gaagcggccg cacttccccc ggagcctccg cttccgccct
     2941 tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca
     3001 tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga ctgggtgctc aggtagtggt
     3061 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga
     3121 gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct
     3181 tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc
     3241 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
     3301 tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga
     3361 tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
     3421 tgtggtcggg gtagcggctg aagcactgca cgccgtaggt cagggtggtc acgagggtgg
     3481 gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg
     3541 tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca
     3601 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg
     3661 cgaccggtgg atcccgggcc cgcggtaccg tcgactgcag aattcggttc actaaacgag
     3721 cccggttcac taaacgagct ctgcttatat agacctccca ccgtacacgc ctaccgccca
     3781 tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc
     3841 aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc
     3901 gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta
     3961 atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc
     4021 aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt
     4081 gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag
     4141 tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt
     4201 tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg
     4261 ggctatgaac taatgacccc gtaattgatt actattaata actagcacgc gttcgaagct
     4321 tgagctcggt cccctcgggg ttgggaggtg ggtctgaaac gataatggtg aatatccctg
     4381 cctaactcta ttcactatag aaagtacagc aaaaactatt cttaaaccta ccaagcctcc
     4441 tactatcatt atgaataatt ttatatacca cagccaattt gttatgttaa accaattcca
     4501 caaacttgcc catttatcta attccaataa ttcttgttca ttcttttctt gctggttttg
     4561 cgattcttca attaaggagt gtattaagct tgtgtaattg ttaatttctc tgtcccactc
     4621 catccaggtc gtgtgattcc aaatctgttc cagagattta ttactccaac tagcattcca
     4681 aggcacagca gtggtgcaaa tgagttttcc agagcaaccc caaatcccca ggagctgttg
     4741 atcctttagg tatctttcca cagccaggat tcttgcctgg agctgcttga tgccccagac
     4801 tgtgagttgc aacagatgct gttgcgcctc aatagccctc agcaaattgt tctgctgctg
     4861 cactatacca gacaataatt gtctggcctg taccgtcagc gtcattgacg ctgcgcccat
     4921 agtgcttcct gctgctccca agaacccaag gaacaaagct cctattccca ctgctctttt
     4981 ttctctctgc accactcttc tctttgcctt ggtgggtgct actcctaatg gttcaatttt
     5041 tactacttta tatttatata attcacttct ccaattgtcc ctcatatctc ctccttatgg
     5101 tgttttacta aacttttcca tgttctaatc ctcatcctgt ctacttgcca cacaatcatc
     5161 acctgccatc tgttttccat aatccctaat gatctttgct tttcttcttg gcactacttt
     5221 tatgtcacta ttatcttgta ttactactgc cccttcacct ttccagagga gctttgctgg
     5281 tcctttccaa agtggatttc tgctgtccct gtaataaacc cgaaaatttt gaatttttgt
     5341 aatttgtttt tgtaattctt tagtttgtat gtctgttgct attatgtcta ctattctttc
     5401 ccctgcactg taccccccaa tccccccttt tcttttaaaa ttgtggatga atactgccat
     5461 ttgtactgct gtcttaagat gttcagcctg atctcttacc tgtcctataa ttttctttaa
     5521 ttctttattc atagattcta ctactccttg actttgggga ttgtagggaa tttccaggtc
     5581 tgaagatcct gctgcttgct gtgctttttt cttacttttg ttttgctctt cctctatctt
     5641 gtctaaagct tccttggtgt cttttatctc tatcctttga tgcacacaat agagggttgc
     5701 tactgtatta tataatgatc taagttcttc tgatcctgtc tgaagggatg gttgtagctg
     5761 tcccagtatt tgtctacagc cttctgatgt ttctaacagg ccagtattaa ctgcgaatcg
     5821 ttctagctcc ctgcttgccc atactatatg ttttaattta tattttttct ttccccctgg
     5881 ccttaaccga attttttccc atcgcgatct aattctcccc cgcttaatac tgacgctctc
     5941 gcacccatct ctctccttct agcctccgct agtcaaaatt tttggcgtac tcaccagtcg
     6001 ccgcccctcg cctcttgccg tgcgcgcttc agcaagccga gtcctgcgtc gagagagctc
     6061 ctctggtttc cctttcgctt tcaagtccct gttcgggcgc cactgctaga gattttccac
     6121 actgactaaa agggtctgag ggatctctag ttaccagagt cacacaacag acgggcacac
     6181 actacttgaa gcactcaagg caagctttat tgaggcttaa gcagtgggtt ccctagttag
     6241 ccagagagct cccaggctca gatctggtct aaccagagag acccagtaca ggcaaaaagc
     6301 agctgcttat atgcaggatc tgagggctcg ccactcccca gtcccgccca ggccacgcct
     6361 cccatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag
     6421 gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc
     6481 gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc
     6541 acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt
     6601 gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat
     6661 gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga ttactattaa
     6721 taactagtca ataatcaatg tcgacccagg tggcactttt cggggaaatg tgcgcggaac
     6781 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6841 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6901 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6961 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     7021 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     7081 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     7141 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     7201 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     7261 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     7321 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     7381 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     7441 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     7501 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     7561 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     7621 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     7681 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     7741 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7801 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7861 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7921 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7981 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     8041 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     8101 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     8161 taagtcgtgt cttaccgg
//
