LOCUS       sequence-408158-        8772 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, PWP1_CCDC63 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8772)
  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 8772)
  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_PWP1_CCDC63
FEATURES             Location/Qualifiers
     source          1..8772
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(258..362)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        393..772
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             774..1031
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1078..1203
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1695..1812
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    2172..2405
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2590..2634
                     /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"
     enhancer        2891..3194
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3195..3398
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3349..3369
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3474..3483
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3480..3482,3483..3485,3486..4196)
                     /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(3525..3546)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3786..3805)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4133..4154
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6182..6201)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6247..6302
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6301..6320)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6663..6679)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6663..6679)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6676..6698)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6687..6703
                     /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)."
     promoter        complement(join(6711..6717,6718..6735,6736..6741))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6756..6777
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6836..7031
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6882..7017
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    7037..7171
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7074..7093)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7128..7147
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7239..7256)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7410..7998)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7490..7509)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8169..8772,1..188,189..257))
                     /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"
ORIGIN
        1 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
       61 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
      121 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
      181 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
      241 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
      301 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
      361 cgcacatttc cccgaaaagt gccacctggg tcgacattga ttattgacta gttattaata
      421 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      481 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      541 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      601 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      661 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      721 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tgggaggcgt
      781 ggcctgggcg ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt
      841 tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
      901 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
      961 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     1021 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc
     1081 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg
     1141 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc
     1201 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg
     1261 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
     1321 gcagttaata ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
     1381 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
     1441 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
     1501 gaggaagagc aaaacaaaag taagaaaaaa gcacagcaag cagcaggatc ttcagacctg
     1561 gaaattccct acaatcccca aagtcaagga gtagtagaat ctatgaataa agaattaaag
     1621 aaaattatag gacaggtaag agatcaggct gaacatctta agacagcagt acaaatggca
     1681 gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     1741 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     1801 attcaaaatt ttcgggttta ttacagggac agcagaaatc cactttggaa aggaccagca
     1861 aagctcctct ggaaaggtga aggggcagta gtaatacaag ataatagtga cataaaagta
     1921 gtgccaagaa gaaaagcaaa gatcattagg gattatggaa aacagatggc aggtgatgat
     1981 tgtgtggcaa gtagacagga tgaggattag aacatggaaa agtttagtaa aacaccataa
     2041 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     2101 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     2161 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     2221 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
     2281 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     2341 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     2401 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
     2461 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
     2521 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
     2581 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
     2641 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
     2701 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
     2761 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggaccgag ctcaagcttc
     2821 gaacgcgtgc tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     2881 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     2941 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     3001 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     3061 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     3121 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     3181 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     3241 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     3301 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     3361 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgggctcgtt
     3421 tagtgaaccg aattctgcag tcgacggtac cgcgggcccg ggatccaccg gtcgccacca
     3481 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     3541 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     3601 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     3661 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     3721 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     3781 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     3841 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     3901 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     3961 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     4021 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     4081 acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     4141 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggcg
     4201 gaagcggagg ctccggggga agtgcggccg cttcccggat gaatcgttcg cgtcaagtca
     4261 cctgcgttgc atgggtccgc tgtggggtag cgaaagagac tcccgataag gtcgaactga
     4321 gcaaagaaga agtgaaacgt ttgattgctg aagccaagga aaaattacag gaagaagggg
     4381 gtggctctga cgaagaagaa acaggttctc caagtgagga cggtatgcag agcgcacgta
     4441 cacaagcccg cccacgtgaa cccctggaag atggggaccc ggaggacgac cgtaccttgg
     4501 atgacgatga gttagcagag tatgacttgg ataaatatga tgaggagggg gacccggatg
     4561 ccgagactct tggagagagc ttgcttggac tgactgtata cggatcgaat gaccaagacc
     4621 cctacgtgac gctgaaggat accgaacaat atgagcgcga ggattttctg attaaaccaa
     4681 gtgataacct gatcgtatgc ggacgtgctg aacaggacca gtgtaattta gaagtacacg
     4741 tgtacaatca agaggaagat tccttttatg tgcatcacga tattctttta agcgcttatc
     4801 cactttcagt tgagtggctg aattttgacc cctctcctga tgattctact ggtaattata
     4861 tcgccgttgg aaatatgacg ccagtgatcg aagtttggga ccttgatatc gtggattcat
     4921 tagaaccagt ttttacgctg ggttctaaac tgtccaaaaa gaagaaaaag aagggaaaaa
     4981 agtcatctag tgctgagggt cacacggatg cggttttaga tctttcgtgg aataaattaa
     5041 tccgtaatgt cttggcctcc gcttccgcag acaatactgt gatcctttgg gatatgtctc
     5101 tgggtaagcc cgccgcatca ttagcagttc acactgacaa agttcaaact ctgcaatttc
     5161 accctttcga ggcgcaaaca ttaatcagcg gaagttatga taaatcagtg gcgttatacg
     5221 actgtcgctc accggatgaa tcgcaccgta tgtggcgctt tagcggacaa attgagcgtg
     5281 taacctggaa tcacttcagt ccgtgccact tcttggacga aatcatttta ttacgtagcc
     5341 agcaaaaatt gtctcacgat gacaaccact ccgtattacg tcagttggag gataagctgc
     5401 gtaagacgac agaggaggct gatatgtatg aaagcaaata cggtgaagtc agcaagacat
     5461 tagacctttt aaagaactcg gtggagaagt tattcaagaa aattaattgc gacgctacca
     5521 aaattttagt tcagttgggt gagacgggga aggtcacgga catcaatctg ccccaatatt
     5581 ttgctattat cgagaaaaaa accaacgacc ttctgttact tgagacctat cgtcgcattt
     5641 tagaagtcga aggcgcagaa gcggagatcc ctcctccatt catcaacccg ttctgggggg
     5701 gttcagcatt gcttaagcct ccagagccaa ttaaggtcat cccacccgtt ctgggagccg
     5761 atccgtttag tgatcgtttg gacgatgtcg aacaaccgtt agaccattcc tccctgcgcc
     5821 agcttgttct ggataattac atcctgaagg agaatcgctc gaaggaagta cgcggtgata
     5881 gcttgcctga aaaggtagat gatttccgca gccgcaagaa agtgaccatg tgaaccggtt
     5941 ctagataaga ttacggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     6001 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaaag aagacaagat
     6061 ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     6121 ggctaactag ggaacccact gcttaagcct caataaagct tcagctgctc gagctagcag
     6181 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     6241 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     6301 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt
     6361 agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag
     6421 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga
     6481 cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa
     6541 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca
     6601 tagctgtccc tcttctctta tggagatccc tcgacctgca gcccaagctt ggcgtaatca
     6661 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6721 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6781 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc
     6841 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     6901 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     6961 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     7021 gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     7081 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     7141 actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg
     7201 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     7261 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     7321 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     7381 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     7441 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     7501 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     7561 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     7621 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     7681 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     7741 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     7801 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     7861 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7921 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7981 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     8041 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     8101 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     8161 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     8221 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     8281 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     8341 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     8401 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     8461 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     8521 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     8581 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     8641 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     8701 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     8761 ccgagttgct ct
//
