LOCUS       sequence-407462-        7611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, RARA_WIPF2 from patient-derived
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7611)
  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 7611)
  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_RARA_WIPF2
FEATURES             Location/Qualifiers
     source          1..7611
                     /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(2372..3082,3083..3085,3086..3088))
                     /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(2414..2435)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2763..2782
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3022..3043
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3085..3094
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3170..3373)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3199..3219)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3374..3677
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3934..3978)
                     /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    4163..4396
                     /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    4756..4873
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5365..5490
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5537..5794
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5796..6175
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6206..6310
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6311..6379,6380..7171)
                     /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(6529..6548)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7342..7611,1..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 ccggttcaac
     1801 gaaggatggg tggaagagga ggtgccccgc gtgctgcgcg atttgtcttt gaaggataaa
     1861 tacgttggaa gtgtttgtac tcctcaggcg ctgggaagtc ttcgacggga tgaaatgagt
     1921 atttggattc aaaatcatcc agaaatgagc gcacggttgt aatagaatca cgatgtccgt
     1981 tgcgcaatgg cggcgggggt ggaggaggag gacctgcagg tgtacgtgac ggtgggggcg
     2041 gcggcttacc gcgcgagggt ggttctgacc catgcatacg gtatggtgga ggaggtggcg
     2101 gcgcatcgcg agcaccgttg cggatgaccg gaggcggcgg aggcggagtg gcgggtgatg
     2161 gggtagagta tcccgagacc ggcaactgat gttgtaaagt tgttaaggca cctggcggag
     2221 acaatccgcc cagcatagga gggaaaaaga aagcgtaggg cggcacaggg tatccgttta
     2281 aatgtccacc tccaggagtt gggcagctac tactgttgga ggccattccc cgggaagcgg
     2341 ccgcacttcc cccggagcct ccgcttccgc ccttgtacag ctcgtccatg ccgagagtga
     2401 tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt
     2461 tgctcagctt ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc
     2521 cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt
     2581 ggcggatctt gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt
     2641 tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt
     2701 cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc
     2761 gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg
     2821 gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact
     2881 gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg
     2941 tgcagatgaa cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca
     3001 cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg
     3061 tgaacagctc ctcgcccttg ctcaccatgg tggcgaccgg tggatcccgg gcccgcggta
     3121 ccgtcgactg cagaattcgg ttcactaaac gagcccggtt cactaaacga gctctgctta
     3181 tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac
     3241 gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg
     3301 ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc
     3361 aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     3421 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     3481 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     3541 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     3601 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     3661 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     3721 attactatta ataactagca cgcgttcgaa gcttgagctc ggtcccctcg gggttgggag
     3781 gtgggtctga aacgataatg gtgaatatcc ctgcctaact ctattcacta tagaaagtac
     3841 agcaaaaact attcttaaac ctaccaagcc tcctactatc attatgaata attttatata
     3901 ccacagccaa tttgttatgt taaaccaatt ccacaaactt gcccatttat ctaattccaa
     3961 taattcttgt tcattctttt cttgctggtt ttgcgattct tcaattaagg agtgtattaa
     4021 gcttgtgtaa ttgttaattt ctctgtccca ctccatccag gtcgtgtgat tccaaatctg
     4081 ttccagagat ttattactcc aactagcatt ccaaggcaca gcagtggtgc aaatgagttt
     4141 tccagagcaa ccccaaatcc ccaggagctg ttgatccttt aggtatcttt ccacagccag
     4201 gattcttgcc tggagctgct tgatgcccca gactgtgagt tgcaacagat gctgttgcgc
     4261 ctcaatagcc ctcagcaaat tgttctgctg ctgcactata ccagacaata attgtctggc
     4321 ctgtaccgtc agcgtcattg acgctgcgcc catagtgctt cctgctgctc ccaagaaccc
     4381 aaggaacaaa gctcctattc ccactgctct tttttctctc tgcaccactc ttctctttgc
     4441 cttggtgggt gctactccta atggttcaat ttttactact ttatatttat ataattcact
     4501 tctccaattg tccctcatat ctcctcctta tggtgtttta ctaaactttt ccatgttcta
     4561 atcctcatcc tgtctacttg ccacacaatc atcacctgcc atctgttttc cataatccct
     4621 aatgatcttt gcttttcttc ttggcactac ttttatgtca ctattatctt gtattactac
     4681 tgccccttca cctttccaga ggagctttgc tggtcctttc caaagtggat ttctgctgtc
     4741 cctgtaataa acccgaaaat tttgaatttt tgtaatttgt ttttgtaatt ctttagtttg
     4801 tatgtctgtt gctattatgt ctactattct ttcccctgca ctgtaccccc caatcccccc
     4861 ttttctttta aaattgtgga tgaatactgc catttgtact gctgtcttaa gatgttcagc
     4921 ctgatctctt acctgtccta taattttctt taattcttta ttcatagatt ctactactcc
     4981 ttgactttgg ggattgtagg gaatttccag gtctgaagat cctgctgctt gctgtgcttt
     5041 tttcttactt ttgttttgct cttcctctat cttgtctaaa gcttccttgg tgtcttttat
     5101 ctctatcctt tgatgcacac aatagagggt tgctactgta ttatataatg atctaagttc
     5161 ttctgatcct gtctgaaggg atggttgtag ctgtcccagt atttgtctac agccttctga
     5221 tgtttctaac aggccagtat taactgcgaa tcgttctagc tccctgcttg cccatactat
     5281 atgttttaat ttatattttt tctttccccc tggccttaac cgaatttttt cccatcgcga
     5341 tctaattctc ccccgcttaa tactgacgct ctcgcaccca tctctctcct tctagcctcc
     5401 gctagtcaaa atttttggcg tactcaccag tcgccgcccc tcgcctcttg ccgtgcgcgc
     5461 ttcagcaagc cgagtcctgc gtcgagagag ctcctctggt ttccctttcg ctttcaagtc
     5521 cctgttcggg cgccactgct agagattttc cacactgact aaaagggtct gagggatctc
     5581 tagttaccag agtcacacaa cagacgggca cacactactt gaagcactca aggcaagctt
     5641 tattgaggct taagcagtgg gttccctagt tagccagaga gctcccaggc tcagatctgg
     5701 tctaaccaga gagacccagt acaggcaaaa agcagctgct tatatgcagg atctgagggc
     5761 tcgccactcc ccagtcccgc ccaggccacg cctcccatgg taatagcgat gactaatacg
     5821 tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg
     5881 ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt
     5941 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
     6001 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
     6061 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta
     6121 tgaactaatg accccgtaat tgattactat taataactag tcaataatca atgtcgaccc
     6181 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     6241 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6301 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     6361 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     6421 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     6481 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     6541 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     6601 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     6661 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     6721 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     6781 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6841 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6901 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6961 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     7021 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     7081 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     7141 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     7201 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     7261 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7321 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7381 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7441 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     7501 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7561 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg g
//
