LOCUS       sequence-407438-        7518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, NSD1_ZNF346 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7518)
  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 7518)
  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_NSD1_ZNF346
FEATURES             Location/Qualifiers
     source          1..7518
                     /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(2279..2989,2990..2992,2993..2995))
                     /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(2321..2342)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2670..2689
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2929..2950
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2992..3001
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3077..3280)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3106..3126)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3281..3584
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3841..3885)
                     /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    4070..4303
                     /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    4663..4780
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5272..5397
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5444..5701
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5703..6082
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6113..6217
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6218..6286,6287..7078)
                     /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(6436..6455)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7249..7518,1..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 cctctaatgt ggtcgaatcc ttctgaatag gttgacgttg catgggaatc tgacctaagg
     1861 agcttgctac cgtcttaggg gactgatttt tgtgcttgaa gcctgagacg tgagcttgat
     1921 actgttcgat tgagttaaga acaattttac aagttttgca tgggtagccc ttcccagcct
     1981 tcatcttcga gtgtgtgtta atcagtggat cgctacaaat gcggcagggc caccaaggac
     2041 ggcgtttgaa tttcgcccaa atcagatcgc cgacctcgta ttttaatggg gtcgatttct
     2101 tctttggctg gcagaagggt aattcttggg aggtactaga actcgaagta ccaggcagct
     2161 ccagcatatt ccctaaggta cttgtagagc tgtcgggatc atctgacaat gcggtgcgcg
     2221 tttttaacgg cattccccgg gaagcggccg cacttccccc ggagcctccg cttccgccct
     2281 tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca
     2341 tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga ctgggtgctc aggtagtggt
     2401 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga
     2461 gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct
     2521 tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc
     2581 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
     2641 tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga
     2701 tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
     2761 tgtggtcggg gtagcggctg aagcactgca cgccgtaggt cagggtggtc acgagggtgg
     2821 gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg
     2881 tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca
     2941 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg
     3001 cgaccggtgg atcccgggcc cgcggtaccg tcgactgcag aattcggttc actaaacgag
     3061 cccggttcac taaacgagct ctgcttatat agacctccca ccgtacacgc ctaccgccca
     3121 tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc
     3181 aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc
     3241 gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta
     3301 atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc
     3361 aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt
     3421 gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag
     3481 tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt
     3541 tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg
     3601 ggctatgaac taatgacccc gtaattgatt actattaata actagcacgc gttcgaagct
     3661 tgagctcggt cccctcgggg ttgggaggtg ggtctgaaac gataatggtg aatatccctg
     3721 cctaactcta ttcactatag aaagtacagc aaaaactatt cttaaaccta ccaagcctcc
     3781 tactatcatt atgaataatt ttatatacca cagccaattt gttatgttaa accaattcca
     3841 caaacttgcc catttatcta attccaataa ttcttgttca ttcttttctt gctggttttg
     3901 cgattcttca attaaggagt gtattaagct tgtgtaattg ttaatttctc tgtcccactc
     3961 catccaggtc gtgtgattcc aaatctgttc cagagattta ttactccaac tagcattcca
     4021 aggcacagca gtggtgcaaa tgagttttcc agagcaaccc caaatcccca ggagctgttg
     4081 atcctttagg tatctttcca cagccaggat tcttgcctgg agctgcttga tgccccagac
     4141 tgtgagttgc aacagatgct gttgcgcctc aatagccctc agcaaattgt tctgctgctg
     4201 cactatacca gacaataatt gtctggcctg taccgtcagc gtcattgacg ctgcgcccat
     4261 agtgcttcct gctgctccca agaacccaag gaacaaagct cctattccca ctgctctttt
     4321 ttctctctgc accactcttc tctttgcctt ggtgggtgct actcctaatg gttcaatttt
     4381 tactacttta tatttatata attcacttct ccaattgtcc ctcatatctc ctccttatgg
     4441 tgttttacta aacttttcca tgttctaatc ctcatcctgt ctacttgcca cacaatcatc
     4501 acctgccatc tgttttccat aatccctaat gatctttgct tttcttcttg gcactacttt
     4561 tatgtcacta ttatcttgta ttactactgc cccttcacct ttccagagga gctttgctgg
     4621 tcctttccaa agtggatttc tgctgtccct gtaataaacc cgaaaatttt gaatttttgt
     4681 aatttgtttt tgtaattctt tagtttgtat gtctgttgct attatgtcta ctattctttc
     4741 ccctgcactg taccccccaa tccccccttt tcttttaaaa ttgtggatga atactgccat
     4801 ttgtactgct gtcttaagat gttcagcctg atctcttacc tgtcctataa ttttctttaa
     4861 ttctttattc atagattcta ctactccttg actttgggga ttgtagggaa tttccaggtc
     4921 tgaagatcct gctgcttgct gtgctttttt cttacttttg ttttgctctt cctctatctt
     4981 gtctaaagct tccttggtgt cttttatctc tatcctttga tgcacacaat agagggttgc
     5041 tactgtatta tataatgatc taagttcttc tgatcctgtc tgaagggatg gttgtagctg
     5101 tcccagtatt tgtctacagc cttctgatgt ttctaacagg ccagtattaa ctgcgaatcg
     5161 ttctagctcc ctgcttgccc atactatatg ttttaattta tattttttct ttccccctgg
     5221 ccttaaccga attttttccc atcgcgatct aattctcccc cgcttaatac tgacgctctc
     5281 gcacccatct ctctccttct agcctccgct agtcaaaatt tttggcgtac tcaccagtcg
     5341 ccgcccctcg cctcttgccg tgcgcgcttc agcaagccga gtcctgcgtc gagagagctc
     5401 ctctggtttc cctttcgctt tcaagtccct gttcgggcgc cactgctaga gattttccac
     5461 actgactaaa agggtctgag ggatctctag ttaccagagt cacacaacag acgggcacac
     5521 actacttgaa gcactcaagg caagctttat tgaggcttaa gcagtgggtt ccctagttag
     5581 ccagagagct cccaggctca gatctggtct aaccagagag acccagtaca ggcaaaaagc
     5641 agctgcttat atgcaggatc tgagggctcg ccactcccca gtcccgccca ggccacgcct
     5701 cccatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag
     5761 gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc
     5821 gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc
     5881 acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt
     5941 gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat
     6001 gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga ttactattaa
     6061 taactagtca ataatcaatg tcgacccagg tggcactttt cggggaaatg tgcgcggaac
     6121 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6181 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6241 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6301 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6361 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     6421 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     6481 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6541 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6601 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6661 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6721 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6781 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6841 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6901 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6961 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     7021 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     7081 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7141 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7201 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7261 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7321 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7381 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7441 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7501 taagtcgtgt cttaccgg
//
