LOCUS       sequence-408098-        8076 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, KPNA1_GMPS from patient-derived
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8076)
  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 8076)
  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_KPNA1_GMPS
FEATURES             Location/Qualifiers
     source          1..8076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     136..155
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(374..478)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        509..888
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             890..1147
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1194..1319
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1811..1928
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    2288..2521
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2706..2750
                     /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        3007..3310
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3311..3514
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3465..3485
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3590..3599
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3596..3598,3599..3601,3602..4312)
                     /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(3641..3662)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3902..3921)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4249..4270
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5602..5621)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5667..5722
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5721..5740)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6083..6099)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6083..6099)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6096..6118)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6107..6123
                     /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(6131..6137,6138..6155,6156..6161))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6176..6197
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6256..6451
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6302..6437
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6457..6591
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6494..6513)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6548..6567
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6659..6676)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6830..7418)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6910..6929)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7589..8076,1..304,305..373))
                     /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 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
       61 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
      121 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
      181 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
      241 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
      301 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
      361 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
      421 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
      481 catttccccg aaaagtgcca cctgggtcga cattgattat tgactagtta ttaatagtaa
      541 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      601 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      661 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      721 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      781 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      841 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggg aggcgtggcc
      901 tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc
      961 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     1021 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     1081 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     1141 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     1201 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     1261 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     1321 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     1381 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     1441 ttaatactgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     1501 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     1561 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     1621 aagagcaaaa caaaagtaag aaaaaagcac agcaagcagc aggatcttca gacctggaaa
     1681 ttccctacaa tccccaaagt caaggagtag tagaatctat gaataaagaa ttaaagaaaa
     1741 ttataggaca ggtaagagat caggctgaac atcttaagac agcagtacaa atggcagtat
     1801 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
     1861 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
     1921 aaaattttcg ggtttattac agggacagca gaaatccact ttggaaagga ccagcaaagc
     1981 tcctctggaa aggtgaaggg gcagtagtaa tacaagataa tagtgacata aaagtagtgc
     2041 caagaagaaa agcaaagatc attagggatt atggaaaaca gatggcaggt gatgattgtg
     2101 tggcaagtag acaggatgag gattagaaca tggaaaagtt tagtaaaaca ccataaggag
     2161 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     2221 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     2281 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     2341 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     2401 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     2461 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     2521 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     2581 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     2641 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
     2701 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
     2761 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     2821 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
     2881 caccattatc gtttcagacc cacctcccaa ccccgagggg accgagctca agcttcgaac
     2941 gcgtgctagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     3001 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     3061 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     3121 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     3181 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     3241 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     3301 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     3361 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     3421 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     3481 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccggg ctcgtttagt
     3541 gaaccgaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggtcg ccaccatggt
     3601 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     3661 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     3721 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     3781 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     3841 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     3901 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     3961 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     4021 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     4081 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     4141 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     4201 gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     4261 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agggcggaag
     4321 cggaggctcc gggggaagtg cggccgcttc ccggatgacc acgccaggta aagaaaattt
     4381 ccgcttgaag agttacaaga acaagagcct taatccggat gaaatgcgcc gtcgccgcga
     4441 agaagagggc ttacaattgc gtaaacagaa gcgcgaagag caaggcaagg ttatcgaacc
     4501 tttgaaggac tttcataagg acgaggtgcg catcctgggg cgtgagttgg gattgcccga
     4561 agagctggtc tcgcgtcatc cgtttcctgg ccccggattg gctatccgcg taatctgcgc
     4621 tgaggagccc tacatctgca aagattttcc agaaacaaac aacattttaa aaatcgtagc
     4681 ggacttctca gcgtcggtga agaaaccgca tactctttta cagcgcgtga aagcatgcac
     4741 gactgaggag gaccaagaaa agttaatgca gatcacgtct ctgcatagct tgaatgcatt
     4801 cttgttaccg atcaaaacgg ttggggtgca gggtgactgc cgttcatact cttacgtgtg
     4861 cggaattagt tccaaagatg aacctgattg ggagagcctt atcttcttag ctcgcttgat
     4921 cccacgtatg tgccacaacg tgaaccgtgt tgtttacatc tttggtccac ccgtgaaaga
     4981 gccgccgaca gacgtaaccc caactttcct gacaactggc gtcttatcta cattacgcca
     5041 ggcggacttc gaggcacaca acattttacg cgaaagtggg tacgccggaa agatttcgca
     5101 aatgccagtt atcttaacac ccttacattt tgaccgcgac ccgcttcaga aacagccgtc
     5161 gtgtcagcgc tctgtcgtta tccgcacctt catcacttcg gacttcatga ctggcattcc
     5221 cgctacacct gggaatgaga tcccagtaga agttgtactg aagatggtta ccgagatcaa
     5281 gaaaatccct ggtatctctc gcattatgta tgacctgact tcaaaacccc ccggaaccac
     5341 tgagtgggaa tgaaccggtt ctagataaga ttacggtacc tttaagacca atgacttaca
     5401 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     5461 actcccaaag aagacaagat ctgctttttg cctgtactgg gtctctctgg ttagaccaga
     5521 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     5581 tcagctgctc gagctagcag atctttttcc ctctgccaaa aattatgggg acatcatgaa
     5641 gccccttgag catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg
     5701 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca
     5761 tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa
     5821 caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt
     5881 attccataga aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt
     5941 tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct
     6001 ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacctgca
     6061 gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6121 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6181 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6241 cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     6301 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     6361 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     6421 aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat
     6481 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6541 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt
     6601 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6661 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6721 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6781 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6841 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6901 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6961 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     7021 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7081 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7141 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7201 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7261 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7321 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7381 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7441 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7501 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7561 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7621 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7681 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7741 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7801 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7861 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7921 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7981 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     8041 gaagtaagtt ggccgcagtg ttatcactca tggtta
//
