LOCUS       sequence-407499-        8577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, WDR37_JAK2 from patient-derived
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8577)
  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 8577)
  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_WDR37_JAK2
FEATURES             Location/Qualifiers
     source          1..8577
                     /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(3338..4048,4049..4051,4052..4054))
                     /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(3380..3401)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3729..3748
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3988..4009
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4051..4060
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4136..4339)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4165..4185)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4340..4643
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4900..4944)
                     /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    5129..5362
                     /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    5722..5839
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6331..6456
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6503..6760
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6762..7141
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7172..7276
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7277..7345,7346..8137)
                     /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(7495..7514)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8308..8577,1..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 ccggttcacc
     1801 cggccatgtt atcacggatt tggtctacac gaagtgccaa gtcgcgaaaa gaaggacgct
     1861 ggtttacatt gttgttccaa cattccgtca taatcatgta aatttcgtcg ggacagccat
     1921 cgggacgggg taaacgtccg ttatttttca ataattcgat caagtgaaat acaatcattt
     1981 gtccttgttt gtcgtttcca atcatgcgca taaattctgc aggcggggat ttggatttct
     2041 caatatatgt gaacagttca tacagcacga ccccgaagct ccaaacgtca ctcgcaacgc
     2101 taaacttaga ctcagtcaaa ctttcgggcg cataccagaa aattgggctc tcgccgggct
     2161 ccttaacctt atagtattcc ttatcttggg gcagaacctt ggtaagtcca aagtccccga
     2221 ttttcacgcg attctcattt tctaccaaaa tattacgagt cgctaaatcg cgatgaatat
     2281 aacgcttggt gcccaggtac tccatacctt tgcagatttg tgatgtatac tgcagtaatt
     2341 tgatatggtc gatacgttct ttgtgtttct gaaggtaatc acgtaagcta ccgtagggta
     2401 aatactccat aataagttta aggttacgac gacccgcaga gtagcatacc cccttgtatt
     2461 taacaatgtt gtcatgctgt aaacttttca agatctcaat ttcacgctca aagtcgcgaa
     2521 gatgctcctc cgtggaatgt tgtaatttct tgacggcgac cacttcgcct gtgttgtcct
     2581 gcaacgggtc ataacggcac atttctactg aaccaaaatt tccttttccc agttgctgta
     2641 aaaatttcaa atgacgctcc tcgaattgtg tagggtcgcg atcttcaaag gcccccgaaa
     2701 atccaagggc cccgatacgc atgtttggca gcatatcgtt ctccgtcagt aattcataat
     2761 ccgcgctcgc cgtacccagc actacgggtt gtgttttggc caccgagaca tcccaaatgc
     2821 cgtcgcggtg cccaatgtac tctttcacta actggcacgc tgcgcgagac gtagtggtct
     2881 tgaagctact cacgattttg gatgtgcttg ccttgtacgt cgtctttaac ttttgggata
     2941 actgggatgt tgaatgagag gctttggttt ttaattgtcc ctttgacaac tcggcaccat
     3001 caatcgcctg gccctccgca gctaaacgtt cgttcaacgt gtcgatttca cggcgcaatt
     3061 ccaggttctc aatgtataaa ttttcaaatt cacgctcaat ctggccgaat aactccaaca
     3121 atgttgagcg tactgagcta ggtaacttgc tatcctggcc ctccaacatg tcgcggggta
     3181 accctgtacg ttcctgttca gaagagttcg tgcgacgaat ggaaagagaa tgagacttac
     3241 gcttctgttt cgtttgacgg gcggtcgagc acgatgcgct ctccgttggc attccccggg
     3301 aagcggccgc acttcccccg gagcctccgc ttccgccctt gtacagctcg tccatgccga
     3361 gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg
     3421 ggtctttgct cagcttggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc
     3481 cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga
     3541 tgttgtggcg gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg gccatgatat
     3601 agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct
     3661 tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct
     3721 cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc
     3781 cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga
     3841 agcactgcac gccgtaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc
     3901 cggtggtgca gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc
     3961 cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca
     4021 ccccggtgaa cagctcctcg cccttgctca ccatggtggc gaccggtgga tcccgggccc
     4081 gcggtaccgt cgactgcaga attcggttca ctaaacgagc ccggttcact aaacgagctc
     4141 tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt
     4201 tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt
     4261 caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt
     4321 actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag
     4381 taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
     4441 tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag
     4501 tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg
     4561 gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc
     4621 catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg
     4681 taattgatta ctattaataa ctagcacgcg ttcgaagctt gagctcggtc ccctcggggt
     4741 tgggaggtgg gtctgaaacg ataatggtga atatccctgc ctaactctat tcactataga
     4801 aagtacagca aaaactattc ttaaacctac caagcctcct actatcatta tgaataattt
     4861 tatataccac agccaatttg ttatgttaaa ccaattccac aaacttgccc atttatctaa
     4921 ttccaataat tcttgttcat tcttttcttg ctggttttgc gattcttcaa ttaaggagtg
     4981 tattaagctt gtgtaattgt taatttctct gtcccactcc atccaggtcg tgtgattcca
     5041 aatctgttcc agagatttat tactccaact agcattccaa ggcacagcag tggtgcaaat
     5101 gagttttcca gagcaacccc aaatccccag gagctgttga tcctttaggt atctttccac
     5161 agccaggatt cttgcctgga gctgcttgat gccccagact gtgagttgca acagatgctg
     5221 ttgcgcctca atagccctca gcaaattgtt ctgctgctgc actataccag acaataattg
     5281 tctggcctgt accgtcagcg tcattgacgc tgcgcccata gtgcttcctg ctgctcccaa
     5341 gaacccaagg aacaaagctc ctattcccac tgctcttttt tctctctgca ccactcttct
     5401 ctttgccttg gtgggtgcta ctcctaatgg ttcaattttt actactttat atttatataa
     5461 ttcacttctc caattgtccc tcatatctcc tccttatggt gttttactaa acttttccat
     5521 gttctaatcc tcatcctgtc tacttgccac acaatcatca cctgccatct gttttccata
     5581 atccctaatg atctttgctt ttcttcttgg cactactttt atgtcactat tatcttgtat
     5641 tactactgcc ccttcacctt tccagaggag ctttgctggt cctttccaaa gtggatttct
     5701 gctgtccctg taataaaccc gaaaattttg aatttttgta atttgttttt gtaattcttt
     5761 agtttgtatg tctgttgcta ttatgtctac tattctttcc cctgcactgt accccccaat
     5821 cccccctttt cttttaaaat tgtggatgaa tactgccatt tgtactgctg tcttaagatg
     5881 ttcagcctga tctcttacct gtcctataat tttctttaat tctttattca tagattctac
     5941 tactccttga ctttggggat tgtagggaat ttccaggtct gaagatcctg ctgcttgctg
     6001 tgcttttttc ttacttttgt tttgctcttc ctctatcttg tctaaagctt ccttggtgtc
     6061 ttttatctct atcctttgat gcacacaata gagggttgct actgtattat ataatgatct
     6121 aagttcttct gatcctgtct gaagggatgg ttgtagctgt cccagtattt gtctacagcc
     6181 ttctgatgtt tctaacaggc cagtattaac tgcgaatcgt tctagctccc tgcttgccca
     6241 tactatatgt tttaatttat attttttctt tccccctggc cttaaccgaa ttttttccca
     6301 tcgcgatcta attctccccc gcttaatact gacgctctcg cacccatctc tctccttcta
     6361 gcctccgcta gtcaaaattt ttggcgtact caccagtcgc cgcccctcgc ctcttgccgt
     6421 gcgcgcttca gcaagccgag tcctgcgtcg agagagctcc tctggtttcc ctttcgcttt
     6481 caagtccctg ttcgggcgcc actgctagag attttccaca ctgactaaaa gggtctgagg
     6541 gatctctagt taccagagtc acacaacaga cgggcacaca ctacttgaag cactcaaggc
     6601 aagctttatt gaggcttaag cagtgggttc cctagttagc cagagagctc ccaggctcag
     6661 atctggtcta accagagaga cccagtacag gcaaaaagca gctgcttata tgcaggatct
     6721 gagggctcgc cactccccag tcccgcccag gccacgcctc ccatggtaat agcgatgact
     6781 aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc
     6841 caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact
     6901 tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa
     6961 gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg
     7021 ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat
     7081 gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt
     7141 cgacccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     7201 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     7261 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     7321 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     7381 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     7441 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     7501 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     7561 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     7621 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     7681 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     7741 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     7801 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     7861 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     7921 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     7981 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     8041 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     8101 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     8161 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     8221 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     8281 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     8341 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     8401 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     8461 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     8521 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgg
//
