LOCUS       sequence-408150-        8553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, PAX5_KANK1 from patient-derived
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8553)
  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 8553)
  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_PAX5_KANK1
FEATURES             Location/Qualifiers
     source          1..8553
                     /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(3314..4024,4025..4027,4028..4030))
                     /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(3356..3377)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3705..3724
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3964..3985
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4027..4036
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4112..4315)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4141..4161)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4316..4619
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4876..4920)
                     /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    5105..5338
                     /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    5698..5815
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6307..6432
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6479..6736
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6738..7117
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7148..7252
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7253..7321,7322..8113)
                     /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(7471..7490)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8284..8553,1..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 ccggttcaat
     1801 cgaagctgcc cctgtgcgtc ggtcccggtg acgtcttccg accgagccta ggcgtccccg
     1861 gagactgtgc ttttgcgaag tttacgtggg catagaggag aaccgctatg tccttatgtc
     1921 cagcttccaa cgcgattgac agggccgttg aaccatcgtt gtcctccaga tgtccattac
     1981 agccaggctg agccaaaagc agttttacga tttccacatg gccgtgttcg gatgcgcaca
     2041 tcagtgcggt gcttccctcg tcgtcctgta tgtttacatc ggctccacaa gcaaggagtc
     2101 ccttgaccat gtctatcctg ccatgtgaga ccgcaagcat gagggctgtt tgcccagcct
     2161 gggaggcttt agcgtttacg tcaccacagc cgaacagttc ttcaacgatt cgcatatctt
     2221 tctctgcttc tacggcagca agcgcggcca gcattatggg cgtataacca gccttgtttt
     2281 ggtgatcaac gttgcacacg tcagcgtcca gcaacaactt aacgatctca aagtttgaat
     2341 gagagacaga gtagtgaaga gccgtattac cgtttccatc ggcgaggttt ataacgtatc
     2401 gcaggacgtc tggagagatt gcttcaaacg ctgctatata atccccgacc atagccggta
     2461 tggcggattt ctgtgagcta actcggaacc attcatgctg gagtgtgttc aaacaaaacc
     2521 tcatatcctt agatgtcaga gccttggggt cgtttatggt attcttgagg agattgcagg
     2581 cagacagcat tttctcgcta agctcgtacc caccgtttat gccgacgaat tgaaggtttt
     2641 tcttggcgcc gtttgagtct ttgttcccat cttttttttt cataatactt ttaagtgttg
     2701 attcattgtt ggtacaggct tcttgagtgg ggaaggcatc cagactactg atgggttttc
     2761 cttcactcgt ccctacttcc tgttccggtt gagacgtttg actgctcagc ggagatcctg
     2821 actgcaaacc cccgcatttg catgtgtatc caaggtaatt gccttcgtct cgcttgcgtt
     2881 tattggtgtc agcagaagga gaagtaatgc ccaggattcc tgaaattgaa tatgaacttc
     2941 cagcgctatc tgtggatacg gaagaaactt gtgtcacaga acctgtgctc actatggagt
     3001 gacttgaagc gggtacaggc tggtttggag gttgctggac cttcgttcta ataatcctac
     3061 cgagaatctt actgacacat ccgtgtgaga ctctcaactg cctagagatg tcgcacgggc
     3121 gtaccccctg atgtgccaac tctactattc tctgtctgac gacatctgga agaggccgcc
     3181 cattgacaaa aactccgccc agttggttaa cgcccccatg cccggttcgg ctggtcctcg
     3241 gagtagggta gtttttctca aggtccattc cccgggaagc ggccgcactt cccccggagc
     3301 ctccgcttcc gcccttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga
     3361 actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagc ttggactggg
     3421 tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct
     3481 ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc ttgaagttca
     3541 ccttgatgcc gttcttctgc ttgtcggcca tgatatagac gttgtggctg ttgtagttgt
     3601 actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga
     3661 tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt
     3721 cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga
     3781 agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg
     3841 tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg
     3901 tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt
     3961 ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct
     4021 tgctcaccat ggtggcgacc ggtggatccc gggcccgcgg taccgtcgac tgcagaattc
     4081 ggttcactaa acgagcccgg ttcactaaac gagctctgct tatatagacc tcccaccgta
     4141 cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc
     4201 gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc
     4261 cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg
     4321 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     4381 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     4441 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     4501 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     4561 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     4621 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat taataactag
     4681 cacgcgttcg aagcttgagc tcggtcccct cggggttggg aggtgggtct gaaacgataa
     4741 tggtgaatat ccctgcctaa ctctattcac tatagaaagt acagcaaaaa ctattcttaa
     4801 acctaccaag cctcctacta tcattatgaa taattttata taccacagcc aatttgttat
     4861 gttaaaccaa ttccacaaac ttgcccattt atctaattcc aataattctt gttcattctt
     4921 ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt aagcttgtgt aattgttaat
     4981 ttctctgtcc cactccatcc aggtcgtgtg attccaaatc tgttccagag atttattact
     5041 ccaactagca ttccaaggca cagcagtggt gcaaatgagt tttccagagc aaccccaaat
     5101 ccccaggagc tgttgatcct ttaggtatct ttccacagcc aggattcttg cctggagctg
     5161 cttgatgccc cagactgtga gttgcaacag atgctgttgc gcctcaatag ccctcagcaa
     5221 attgttctgc tgctgcacta taccagacaa taattgtctg gcctgtaccg tcagcgtcat
     5281 tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac ccaaggaaca aagctcctat
     5341 tcccactgct cttttttctc tctgcaccac tcttctcttt gccttggtgg gtgctactcc
     5401 taatggttca atttttacta ctttatattt atataattca cttctccaat tgtccctcat
     5461 atctcctcct tatggtgttt tactaaactt ttccatgttc taatcctcat cctgtctact
     5521 tgccacacaa tcatcacctg ccatctgttt tccataatcc ctaatgatct ttgcttttct
     5581 tcttggcact acttttatgt cactattatc ttgtattact actgcccctt cacctttcca
     5641 gaggagcttt gctggtcctt tccaaagtgg atttctgctg tccctgtaat aaacccgaaa
     5701 attttgaatt tttgtaattt gtttttgtaa ttctttagtt tgtatgtctg ttgctattat
     5761 gtctactatt ctttcccctg cactgtaccc cccaatcccc ccttttcttt taaaattgtg
     5821 gatgaatact gccatttgta ctgctgtctt aagatgttca gcctgatctc ttacctgtcc
     5881 tataattttc tttaattctt tattcataga ttctactact ccttgacttt ggggattgta
     5941 gggaatttcc aggtctgaag atcctgctgc ttgctgtgct tttttcttac ttttgttttg
     6001 ctcttcctct atcttgtcta aagcttcctt ggtgtctttt atctctatcc tttgatgcac
     6061 acaatagagg gttgctactg tattatataa tgatctaagt tcttctgatc ctgtctgaag
     6121 ggatggttgt agctgtccca gtatttgtct acagccttct gatgtttcta acaggccagt
     6181 attaactgcg aatcgttcta gctccctgct tgcccatact atatgtttta atttatattt
     6241 tttctttccc cctggcctta accgaatttt ttcccatcgc gatctaattc tcccccgctt
     6301 aatactgacg ctctcgcacc catctctctc cttctagcct ccgctagtca aaatttttgg
     6361 cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc gcttcagcaa gccgagtcct
     6421 gcgtcgagag agctcctctg gtttcccttt cgctttcaag tccctgttcg ggcgccactg
     6481 ctagagattt tccacactga ctaaaagggt ctgagggatc tctagttacc agagtcacac
     6541 aacagacggg cacacactac ttgaagcact caaggcaagc tttattgagg cttaagcagt
     6601 gggttcccta gttagccaga gagctcccag gctcagatct ggtctaacca gagagaccca
     6661 gtacaggcaa aaagcagctg cttatatgca ggatctgagg gctcgccact ccccagtccc
     6721 gcccaggcca cgcctcccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
     6781 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
     6841 acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt
     6901 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga
     6961 acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca
     7021 tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta
     7081 attgattact attaataact agtcaataat caatgtcgac ccaggtggca cttttcgggg
     7141 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     7201 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     7261 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     7321 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     7381 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     7441 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     7501 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     7561 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     7621 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     7681 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     7741 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     7801 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     7861 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     7921 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     7981 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     8041 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     8101 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     8161 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     8221 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     8281 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     8341 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     8401 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     8461 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     8521 tgctgccagt ggcgataagt cgtgtcttac cgg
//
