LOCUS       sequence-406610-        7872 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, CLK1_PPIL3 from patient-derived
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7872)
  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 7872)
  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_CLK1_PPIL3
FEATURES             Location/Qualifiers
     source          1..7872
                     /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(2633..3343,3344..3346,3347..3349))
                     /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(2675..2696)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3024..3043
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3283..3304
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3346..3355
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3431..3634)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3460..3480)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3635..3938
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4195..4239)
                     /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    4424..4657
                     /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    5017..5134
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5626..5751
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5798..6055
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6057..6436
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6467..6571
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6572..6640,6641..7432)
                     /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(6790..6809)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7603..7872,1..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 ccggttcact
     1801 gcgcaaaagg gtttgcgtgg atcgtaatgt ctttgatatg gacatcattc aatgggcggt
     1861 aagtcttttc attaactgga agtttttcta attcatccaa ggtttccagc ccgtcgatca
     1921 ctaacgtgct attcgggtcc gtagtgttaa gatgttctaa aacttgaatt tcgctacgtg
     1981 cagcctcaca atagcgatca acatttttca cgatttttac agccacatgg cgtcccccag
     2041 ctttgtgatc gatgcactca accacctttc caaaggctcc ctcgcccaaa gtatcgacga
     2101 tctcgtagcg ggcagaaaga acgtcacctg attgacagat cagatgcccc tcttcatcat
     2161 cttcaacgct acgagtgcgt ttgcgacggt gagacttacc atgcgaacgg cggtggcttg
     2221 tgctatgatg gatgcggtgc ttagatttgt acgatgagcg ccctgagcgg ccagacgact
     2281 tagatgaatg attctgatag cgactttcat ggtcacgttg gcgatgtcct ggttcgcacc
     2341 cctgtgtgta gtcgttgcga tactcatcaa tatagcgacg agaatgatag tccttttcat
     2401 tgatcgaacg agactccaaa taatgagaat cgcacatctt gctgtgatta tatttacaac
     2461 gcttgttctc ttgcgccgaa ctatgagaac gtttacgacg tttgtgggaa gagctggagc
     2521 gccacttccc gtagtcccaa tctttgtcgt cccaatcggg acaatatgtg cgtttagaat
     2581 gacgcattcc ccgggaagcg gccgcacttc ccccggagcc tccgcttccg cccttgtaca
     2641 gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat
     2701 cgcgcttctc gttggggtct ttgctcagct tggactgggt gctcaggtag tggttgtcgg
     2761 gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca
     2821 cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct
     2881 tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga
     2941 tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc
     3001 cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc
     3061 gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt
     3121 cggggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg
     3181 gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat
     3241 cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga
     3301 ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcgaccg
     3361 gtggatcccg ggcccgcggt accgtcgact gcagaattcg gttcactaaa cgagcccggt
     3421 tcactaaacg agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg
     3481 tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca
     3541 aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc
     3601 cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt
     3661 agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg
     3721 gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta
     3781 ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta
     3841 ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg
     3901 gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat
     3961 gaactaatga ccccgtaatt gattactatt aataactagc acgcgttcga agcttgagct
     4021 cggtcccctc ggggttggga ggtgggtctg aaacgataat ggtgaatatc cctgcctaac
     4081 tctattcact atagaaagta cagcaaaaac tattcttaaa cctaccaagc ctcctactat
     4141 cattatgaat aattttatat accacagcca atttgttatg ttaaaccaat tccacaaact
     4201 tgcccattta tctaattcca ataattcttg ttcattcttt tcttgctggt tttgcgattc
     4261 ttcaattaag gagtgtatta agcttgtgta attgttaatt tctctgtccc actccatcca
     4321 ggtcgtgtga ttccaaatct gttccagaga tttattactc caactagcat tccaaggcac
     4381 agcagtggtg caaatgagtt ttccagagca accccaaatc cccaggagct gttgatcctt
     4441 taggtatctt tccacagcca ggattcttgc ctggagctgc ttgatgcccc agactgtgag
     4501 ttgcaacaga tgctgttgcg cctcaatagc cctcagcaaa ttgttctgct gctgcactat
     4561 accagacaat aattgtctgg cctgtaccgt cagcgtcatt gacgctgcgc ccatagtgct
     4621 tcctgctgct cccaagaacc caaggaacaa agctcctatt cccactgctc ttttttctct
     4681 ctgcaccact cttctctttg ccttggtggg tgctactcct aatggttcaa tttttactac
     4741 tttatattta tataattcac ttctccaatt gtccctcata tctcctcctt atggtgtttt
     4801 actaaacttt tccatgttct aatcctcatc ctgtctactt gccacacaat catcacctgc
     4861 catctgtttt ccataatccc taatgatctt tgcttttctt cttggcacta cttttatgtc
     4921 actattatct tgtattacta ctgccccttc acctttccag aggagctttg ctggtccttt
     4981 ccaaagtgga tttctgctgt ccctgtaata aacccgaaaa ttttgaattt ttgtaatttg
     5041 tttttgtaat tctttagttt gtatgtctgt tgctattatg tctactattc tttcccctgc
     5101 actgtacccc ccaatccccc cttttctttt aaaattgtgg atgaatactg ccatttgtac
     5161 tgctgtctta agatgttcag cctgatctct tacctgtcct ataattttct ttaattcttt
     5221 attcatagat tctactactc cttgactttg gggattgtag ggaatttcca ggtctgaaga
     5281 tcctgctgct tgctgtgctt ttttcttact tttgttttgc tcttcctcta tcttgtctaa
     5341 agcttccttg gtgtctttta tctctatcct ttgatgcaca caatagaggg ttgctactgt
     5401 attatataat gatctaagtt cttctgatcc tgtctgaagg gatggttgta gctgtcccag
     5461 tatttgtcta cagccttctg atgtttctaa caggccagta ttaactgcga atcgttctag
     5521 ctccctgctt gcccatacta tatgttttaa tttatatttt ttctttcccc ctggccttaa
     5581 ccgaattttt tcccatcgcg atctaattct cccccgctta atactgacgc tctcgcaccc
     5641 atctctctcc ttctagcctc cgctagtcaa aatttttggc gtactcacca gtcgccgccc
     5701 ctcgcctctt gccgtgcgcg cttcagcaag ccgagtcctg cgtcgagaga gctcctctgg
     5761 tttccctttc gctttcaagt ccctgttcgg gcgccactgc tagagatttt ccacactgac
     5821 taaaagggtc tgagggatct ctagttacca gagtcacaca acagacgggc acacactact
     5881 tgaagcactc aaggcaagct ttattgaggc ttaagcagtg ggttccctag ttagccagag
     5941 agctcccagg ctcagatctg gtctaaccag agagacccag tacaggcaaa aagcagctgc
     6001 ttatatgcag gatctgaggg ctcgccactc cccagtcccg cccaggccac gcctcccatg
     6061 gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg
     6121 tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt
     6181 ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat
     6241 tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc
     6301 aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg
     6361 cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta
     6421 gtcaataatc aatgtcgacc caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     6481 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6541 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     6601 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6661 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     6721 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     6781 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6841 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6901 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6961 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     7021 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     7081 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     7141 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     7201 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     7261 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     7321 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     7381 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     7441 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     7501 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     7561 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     7621 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     7681 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     7741 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     7801 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7861 gtgtcttacc gg
//
