LOCUS       sequence-407505-        8466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, EEF2_SMARCA4 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8466)
  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 8466)
  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_EEF2_SMARCA4
FEATURES             Location/Qualifiers
     source          1..8466
                     /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(3227..3937,3938..3940,3941..3943))
                     /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(3269..3290)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3618..3637
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3877..3898
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3940..3949
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4025..4228)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4054..4074)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4229..4532
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4789..4833)
                     /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    5018..5251
                     /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    5611..5728
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6220..6345
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6392..6649
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6651..7030
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7061..7165
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7166..7234,7235..8026)
                     /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(7384..7403)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8197..8466,1..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 ccggttcaat
     1801 cttcctcgct cccactgcca ctacggtcct cttcttgctc ttcttcagag tcgtcatccg
     1861 acacgacagg cttcgcacgt gacccgcggg aagggcgacg acggcctcct ttaagacgat
     1921 cctgtgcttt ttctttgcga cccaatttga ttttaacttt aacggagcga ctttcagatt
     1981 cactcccctc ctcctctcct tcttcctcct cctcagattc ctcaccttcg ctgtcgtcct
     2041 ctttctcgat cttttgacgc acgctagtga aaacggattg cagtacgata gaatcttcat
     2101 aaatcaatga cccctcaagg ttaaacgttt gcgcattctg gcataataac ataacatcct
     2161 tttccaggtc gttcaacgaa cggtatttgt gattgcggat gcgttcctta atttttttga
     2221 agtccaccgg cttacggatc agttcgtagt actcaggcaa ctctttgcgc gagggtaatt
     2281 ggataaagac ttccgacagt tggcgtcccg atgacgagtc cttgtactta atcacagcat
     2341 ccacaatctt cttcattttt ttagttaaat tcggtggatt cggagataac ttctctgcgg
     2401 gcgggcgtcc acgcttcttc tgttttttgg attcatcatc tttatcacgt gagcgggtgg
     2461 atgttgttgg ggtgctggac ccggcgtcgc tatcacgttt gcgtttgcgg cttgattttt
     2521 tttggcggac ttcctcttcg atctcttcta acgtgccttc ttcaatcgcc atgatgtttc
     2581 ccataggacc gctctcgcct tcgccgtaag tggagatgat aacattcacg ttttctacaa
     2641 tacgttggaa ggtctgatac aactcttcgg gctcaagctg caactcaagt aatgcgcggt
     2701 ccattttgtt catcattaaa acgggcttga tacgttctgc aatggcttgg cgcaagacgg
     2761 tctctgtctg tacacaaaca ccggaaacac agtccacaac aacaagcgca ccatctgtga
     2821 cgcgcaatgc agctgtaact tcacttgaga agtctacgtg tcccggcgaa tcgattaagt
     2881 taatcaaaaa tccggccccg tccttgcttt gcttgatgaa gttcaaatcg ttctcagaca
     2941 gctcgtagaa caatgagata gcggtcgatt tgattgtaat gcagcgttct tgctcgtctt
     3001 tgcgtgtatc ggtgaagcgt gtctccccgg cgcgggcact ggcgataatc ccagctttgc
     3061 acactaaact atctgtcagg gttgactttc catgatcaac atgggcaatg actgacatat
     3121 tacgaatgtt agctttctta tccatgatag cacgaatctg atctactgtg aaattaacca
     3181 ttccccggga agcggccgca cttcccccgg agcctccgct tccgcccttg tacagctcgt
     3241 ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct
     3301 tctcgttggg gtctttgctc agcttggact gggtgctcag gtagtggttg tcgggcagca
     3361 gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc
     3421 cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg
     3481 ccatgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc
     3541 cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga
     3601 acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct
     3661 ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt
     3721 agcggctgaa gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg
     3781 gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct
     3841 cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga
     3901 tgggcaccac cccggtgaac agctcctcgc ccttgctcac catggtggcg accggtggat
     3961 cccgggcccg cggtaccgtc gactgcagaa ttcggttcac taaacgagcc cggttcacta
     4021 aacgagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
     4081 gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc
     4141 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
     4201 cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt
     4261 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     4321 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     4381 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     4441 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     4501 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     4561 atgaccccgt aattgattac tattaataac tagcacgcgt tcgaagcttg agctcggtcc
     4621 cctcggggtt gggaggtggg tctgaaacga taatggtgaa tatccctgcc taactctatt
     4681 cactatagaa agtacagcaa aaactattct taaacctacc aagcctccta ctatcattat
     4741 gaataatttt atataccaca gccaatttgt tatgttaaac caattccaca aacttgccca
     4801 tttatctaat tccaataatt cttgttcatt cttttcttgc tggttttgcg attcttcaat
     4861 taaggagtgt attaagcttg tgtaattgtt aatttctctg tcccactcca tccaggtcgt
     4921 gtgattccaa atctgttcca gagatttatt actccaacta gcattccaag gcacagcagt
     4981 ggtgcaaatg agttttccag agcaacccca aatccccagg agctgttgat cctttaggta
     5041 tctttccaca gccaggattc ttgcctggag ctgcttgatg ccccagactg tgagttgcaa
     5101 cagatgctgt tgcgcctcaa tagccctcag caaattgttc tgctgctgca ctataccaga
     5161 caataattgt ctggcctgta ccgtcagcgt cattgacgct gcgcccatag tgcttcctgc
     5221 tgctcccaag aacccaagga acaaagctcc tattcccact gctctttttt ctctctgcac
     5281 cactcttctc tttgccttgg tgggtgctac tcctaatggt tcaattttta ctactttata
     5341 tttatataat tcacttctcc aattgtccct catatctcct ccttatggtg ttttactaaa
     5401 cttttccatg ttctaatcct catcctgtct acttgccaca caatcatcac ctgccatctg
     5461 ttttccataa tccctaatga tctttgcttt tcttcttggc actactttta tgtcactatt
     5521 atcttgtatt actactgccc cttcaccttt ccagaggagc tttgctggtc ctttccaaag
     5581 tggatttctg ctgtccctgt aataaacccg aaaattttga atttttgtaa tttgtttttg
     5641 taattcttta gtttgtatgt ctgttgctat tatgtctact attctttccc ctgcactgta
     5701 ccccccaatc cccccttttc ttttaaaatt gtggatgaat actgccattt gtactgctgt
     5761 cttaagatgt tcagcctgat ctcttacctg tcctataatt ttctttaatt ctttattcat
     5821 agattctact actccttgac tttggggatt gtagggaatt tccaggtctg aagatcctgc
     5881 tgcttgctgt gcttttttct tacttttgtt ttgctcttcc tctatcttgt ctaaagcttc
     5941 cttggtgtct tttatctcta tcctttgatg cacacaatag agggttgcta ctgtattata
     6001 taatgatcta agttcttctg atcctgtctg aagggatggt tgtagctgtc ccagtatttg
     6061 tctacagcct tctgatgttt ctaacaggcc agtattaact gcgaatcgtt ctagctccct
     6121 gcttgcccat actatatgtt ttaatttata ttttttcttt ccccctggcc ttaaccgaat
     6181 tttttcccat cgcgatctaa ttctcccccg cttaatactg acgctctcgc acccatctct
     6241 ctccttctag cctccgctag tcaaaatttt tggcgtactc accagtcgcc gcccctcgcc
     6301 tcttgccgtg cgcgcttcag caagccgagt cctgcgtcga gagagctcct ctggtttccc
     6361 tttcgctttc aagtccctgt tcgggcgcca ctgctagaga ttttccacac tgactaaaag
     6421 ggtctgaggg atctctagtt accagagtca cacaacagac gggcacacac tacttgaagc
     6481 actcaaggca agctttattg aggcttaagc agtgggttcc ctagttagcc agagagctcc
     6541 caggctcaga tctggtctaa ccagagagac ccagtacagg caaaaagcag ctgcttatat
     6601 gcaggatctg agggctcgcc actccccagt cccgcccagg ccacgcctcc catggtaata
     6661 gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg
     6721 gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata
     6781 tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt
     6841 caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg
     6901 cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac
     6961 tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata actagtcaat
     7021 aatcaatgtc gacccaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     7081 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     7141 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     7201 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     7261 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     7321 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     7381 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     7441 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     7501 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     7561 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     7621 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     7681 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     7741 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     7801 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     7861 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     7921 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     7981 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     8041 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     8101 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     8161 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     8221 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     8281 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     8341 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     8401 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     8461 taccgg
//
