LOCUS       sequence-408155-        8850 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, PPFIA1_PAPD5 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8850)
  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 8850)
  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_PPFIA1_PAPD5
FEATURES             Location/Qualifiers
     source          1..8850
                     /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(3611..4321,4322..4324,4325..4327))
                     /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(3653..3674)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4002..4021
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4261..4282
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4324..4333
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4409..4612)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4438..4458)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4613..4916
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(5173..5217)
                     /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    5402..5635
                     /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    5995..6112
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6604..6729
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6776..7033
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        7035..7414
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7445..7549
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7550..7618,7619..8410)
                     /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(7768..7787)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8581..8850,1..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 ccggttcagc
     1801 ggcacagatc cgataaaggc gcgtcacgct tatgcttacg cttcttcccg tggtaatact
     1861 gattgttcga ctttccttga tgttgtttat tggtcatcag tgagccatgc gaagtttggg
     1921 ttgtaccttg aaatcccttg ctactggagc ggaacaggcg cgcagagccg tgctgagatt
     1981 tattggtgct gttcgatgta ttagtggttt gggtagattg catttttcca actgcctgtg
     2041 agctctcaag gctaacatcc tggctgccaa cgcgattacc agtagacggg cgacacagaa
     2101 gctgttttgg tgtcttgcac ggagtcgcgt ctgagtctac atcagacgac gagctctgcg
     2161 ttgcgctaga ggaagatacg gggccgcttg atgaatttga ggaatgcttc gacagcgact
     2221 cacttgtttt agagcgacat ttgcccaaag cctcgttctc ctcagacaaa ttattattac
     2281 acttgtctaa ctgttgagtg tccacaatta aggttacacc gttgccgtta caacttggct
     2341 ctggacgatt cttaagcccc cactgtttag aaatccagtc acggtaagtg gctacttcgt
     2401 cagttacacg gataatgcga cctaagatgg actcggtttc attgttggga tagtatttag
     2461 cgatagggct aacagcatgc gacaagacta cataggcgta gtcaaaagcc tgtttaacct
     2521 gcatcgcacc atagctcgaa cggccaacgt cattcccagg ctgcaagggg tcttcgatgt
     2581 acaacatact agggcggtaa ccgtccaaca tgttcttttg gacttcatct ttagcgacgt
     2641 acgacccccc atctttgatg cgaatacctg tcttaagata gttgaagtgg cgtccataaa
     2701 gttcaaagaa ctcaatcaat aaaacgccat aattagtgtt cgggatacaa gcgtcctcgc
     2761 gaggatgaag ttgaagaaaa cttaccgcca tcaggaataa ggaatacgat ccgatgccgc
     2821 cagtaaaaac ttcattcaaa tcacgttgaa gcagaaactg cttcaacact aaaaccagat
     2881 atggcagtac tggatatttt ttagtaaagt ccttaatcag atcagccgcg cgcacgccgt
     2941 tttgaacatt gaaggagatg tcgactttca cttcggtaaa cgaatccgtt aacttaatga
     3001 ttggtacggt agccttatca agaactttca ccgaatcctc atcggctacc ttatgcttac
     3061 gaagagcttc ttcaagcgtc caaagaggca ggttctccca cttgccgaac acaaccaggt
     3121 caatgtcact tgtaggcagg tataagcccg tcttgaaaga cccaaagatt tgaacatccg
     3181 cagacggcca aagttcctta atcactgatt caatgcgatt aactacttcc atgcgcattt
     3241 tctcttcttc tggacgaggt gacatgtact catagaaatc cgagatttct tcatgcaggc
     3301 cctgcggtaa ggcggtgtta agctggcgct gtaaggaatc acgttcgtgg ccaacctcat
     3361 gaagtttacc ttgagttaag gcaagcgtct cttgtgtttc acgaagtgtg tctaaaagac
     3421 ggtcgcgctc ttccaacatg gataccatta attgttcgaa atggctatcc gcatctggtt
     3481 ggcttggact cccagacccg tgccctccac cgccgccagg gggaccctca gcttccgaaa
     3541 tcgtaggcat cacctcgcac atcattcccc gggaagcggc cgcacttccc ccggagcctc
     3601 cgcttccgcc cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact
     3661 ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagcttg gactgggtgc
     3721 tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt
     3781 agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg aagttcacct
     3841 tgatgccgtt cttctgcttg tcggccatga tatagacgtt gtggctgttg tagttgtact
     3901 ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc
     3961 ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt
     4021 ccttgaagaa gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt
     4081 cgtgctgctt catgtggtcg gggtagcggc tgaagcactg cacgccgtag gtcagggtgg
     4141 tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca
     4201 gcttgccgta ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta
     4261 cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc
     4321 tcaccatggt ggcgaccggt ggatcccggg cccgcggtac cgtcgactgc agaattcggt
     4381 tcactaaacg agcccggttc actaaacgag ctctgcttat atagacctcc caccgtacac
     4441 gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt
     4501 gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc
     4561 gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa
     4621 tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact
     4681 gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca
     4741 tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac
     4801 gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg
     4861 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga
     4921 actccatata tgggctatga actaatgacc ccgtaattga ttactattaa taactagcac
     4981 gcgttcgaag cttgagctcg gtcccctcgg ggttgggagg tgggtctgaa acgataatgg
     5041 tgaatatccc tgcctaactc tattcactat agaaagtaca gcaaaaacta ttcttaaacc
     5101 taccaagcct cctactatca ttatgaataa ttttatatac cacagccaat ttgttatgtt
     5161 aaaccaattc cacaaacttg cccatttatc taattccaat aattcttgtt cattcttttc
     5221 ttgctggttt tgcgattctt caattaagga gtgtattaag cttgtgtaat tgttaatttc
     5281 tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt tccagagatt tattactcca
     5341 actagcattc caaggcacag cagtggtgca aatgagtttt ccagagcaac cccaaatccc
     5401 caggagctgt tgatccttta ggtatctttc cacagccagg attcttgcct ggagctgctt
     5461 gatgccccag actgtgagtt gcaacagatg ctgttgcgcc tcaatagccc tcagcaaatt
     5521 gttctgctgc tgcactatac cagacaataa ttgtctggcc tgtaccgtca gcgtcattga
     5581 cgctgcgccc atagtgcttc ctgctgctcc caagaaccca aggaacaaag ctcctattcc
     5641 cactgctctt ttttctctct gcaccactct tctctttgcc ttggtgggtg ctactcctaa
     5701 tggttcaatt tttactactt tatatttata taattcactt ctccaattgt ccctcatatc
     5761 tcctccttat ggtgttttac taaacttttc catgttctaa tcctcatcct gtctacttgc
     5821 cacacaatca tcacctgcca tctgttttcc ataatcccta atgatctttg cttttcttct
     5881 tggcactact tttatgtcac tattatcttg tattactact gccccttcac ctttccagag
     5941 gagctttgct ggtcctttcc aaagtggatt tctgctgtcc ctgtaataaa cccgaaaatt
     6001 ttgaattttt gtaatttgtt tttgtaattc tttagtttgt atgtctgttg ctattatgtc
     6061 tactattctt tcccctgcac tgtacccccc aatcccccct tttcttttaa aattgtggat
     6121 gaatactgcc atttgtactg ctgtcttaag atgttcagcc tgatctctta cctgtcctat
     6181 aattttcttt aattctttat tcatagattc tactactcct tgactttggg gattgtaggg
     6241 aatttccagg tctgaagatc ctgctgcttg ctgtgctttt ttcttacttt tgttttgctc
     6301 ttcctctatc ttgtctaaag cttccttggt gtcttttatc tctatccttt gatgcacaca
     6361 atagagggtt gctactgtat tatataatga tctaagttct tctgatcctg tctgaaggga
     6421 tggttgtagc tgtcccagta tttgtctaca gccttctgat gtttctaaca ggccagtatt
     6481 aactgcgaat cgttctagct ccctgcttgc ccatactata tgttttaatt tatatttttt
     6541 ctttccccct ggccttaacc gaattttttc ccatcgcgat ctaattctcc cccgcttaat
     6601 actgacgctc tcgcacccat ctctctcctt ctagcctccg ctagtcaaaa tttttggcgt
     6661 actcaccagt cgccgcccct cgcctcttgc cgtgcgcgct tcagcaagcc gagtcctgcg
     6721 tcgagagagc tcctctggtt tccctttcgc tttcaagtcc ctgttcgggc gccactgcta
     6781 gagattttcc acactgacta aaagggtctg agggatctct agttaccaga gtcacacaac
     6841 agacgggcac acactacttg aagcactcaa ggcaagcttt attgaggctt aagcagtggg
     6901 ttccctagtt agccagagag ctcccaggct cagatctggt ctaaccagag agacccagta
     6961 caggcaaaaa gcagctgctt atatgcagga tctgagggct cgccactccc cagtcccgcc
     7021 caggccacgc ctcccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga
     7081 aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg
     7141 tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac
     7201 cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca
     7261 tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt
     7321 accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt
     7381 gattactatt aataactagt caataatcaa tgtcgaccca ggtggcactt ttcggggaaa
     7441 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     7501 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     7561 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     7621 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     7681 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     7741 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     7801 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     7861 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     7921 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     7981 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     8041 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     8101 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     8161 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     8221 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     8281 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     8341 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     8401 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     8461 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     8521 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     8581 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     8641 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     8701 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     8761 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     8821 tgccagtggc gataagtcgt gtcttaccgg
//
