LOCUS       sequence-407508-        8490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, DOC2B_USP43 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8490)
  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 8490)
  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_DOC2B_USP43
FEATURES             Location/Qualifiers
     source          1..8490
                     /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(3251..3961,3962..3964,3965..3967))
                     /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(3293..3314)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3642..3661
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3901..3922
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3964..3973
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4049..4252)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4078..4098)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4253..4556
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4813..4857)
                     /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    5042..5275
                     /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    5635..5752
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6244..6369
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6416..6673
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6675..7054
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7085..7189
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7190..7258,7259..8050)
                     /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(7408..7427)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8221..8490,1..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 ccggttcaga
     1801 acgaactctc tggaagagtc tttttgcgtc ccaatgataa cgtgtggtac ttaacgcgct
     1861 gaaatgtccc gtagctagcc tgctcaccgg gaacggtacg ctgggacatt cctaaggcag
     1921 agccacgtgg tgcacgggag gctttgcgcg gaagacgtaa tgagctcggg gcactccaaa
     1981 catcacgctc aaggcgacta ccaagtccac gagccaaccc ctctggaatg cgcagcgccg
     2041 gcggtgaccc aggtgcttgc ccatccattg cgggagaaag acctccagac accaaggata
     2101 ccggtgggac ttgaggtgac acttctgcac gctcgttgcg gcgattttcc tttttgcgaa
     2161 agacactacg cagaagcgta agggtcccct gaaggggacg atctaactca ctcgtaccac
     2221 gacgcgagtc cttagaattg ccggaaaatc ccatgtatgg tgaaggtgga ctgcttttag
     2281 aacccatctg aaatgactcc ttccagttca ttgctttttg cccctcgggg cgcgcaggtt
     2341 tttcatgttc aacactaggc ataacagtca agggcaggtc aaacttacgc ggcagtttaa
     2401 tgtcctgacc tgcattttct ttcagcttac gcgcagtgtt cgggccatca ctattacctg
     2461 gtgcaaggca atgatttggt tgcgcggatg gacatgggac tccggcaggt ccacgctcga
     2521 tagcgcgacc cccgtcctca gaatttgcgg gtaatgcgac attggagcga ggggaggcgg
     2581 atttttcgtc ttctcccgcg gtgcctgtaa gcagggacac gattgattgg gatgtcgaac
     2641 gagggcggcg acgggattcc aagtactcca ccagctcgac gctagcacct ggcggctttt
     2701 ctttcgaccc gaaactccag cgtaatggca tggtcttgaa gggcccctgc tttgcgcgtg
     2761 acgtcgtagg tgccttcatc gaaatcgagc ggcctttaac cccacgaact aaacggcgag
     2821 gctccaggcc gcgacagtca tcggattcat atccgtcggc atcgggttcg tcttcggggg
     2881 gcttagcagg gggacgagca ggagagggac cagggctagg ccctggtgaa gacccgtatg
     2941 ctccgaacag ttggtcaaca tcctcgtcgt cctcacgcgc tccatctgaa gggctacggc
     3001 gaccagctcc ggctacggca ggacgagctg gcgcatctgg gggtgcagcc gcacggggac
     3061 ccgcgtctgg tggcaagcca cggggaaaac gggggaaata atctgaaatt tgtttaatgg
     3121 ggcgaatagg accagggcat acgtcaattg ccatgtgctc ttggatacta atcgttgctt
     3181 tttctccacg gcgacgtaac gtcattcccc gggaagcggc cgcacttccc ccggagcctc
     3241 cgcttccgcc cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact
     3301 ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagcttg gactgggtgc
     3361 tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt
     3421 agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg aagttcacct
     3481 tgatgccgtt cttctgcttg tcggccatga tatagacgtt gtggctgttg tagttgtact
     3541 ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc
     3601 ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt
     3661 ccttgaagaa gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt
     3721 cgtgctgctt catgtggtcg gggtagcggc tgaagcactg cacgccgtag gtcagggtgg
     3781 tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca
     3841 gcttgccgta ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta
     3901 cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc
     3961 tcaccatggt ggcgaccggt ggatcccggg cccgcggtac cgtcgactgc agaattcggt
     4021 tcactaaacg agcccggttc actaaacgag ctctgcttat atagacctcc caccgtacac
     4081 gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt
     4141 gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc
     4201 gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa
     4261 tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact
     4321 gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca
     4381 tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac
     4441 gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg
     4501 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga
     4561 actccatata tgggctatga actaatgacc ccgtaattga ttactattaa taactagcac
     4621 gcgttcgaag cttgagctcg gtcccctcgg ggttgggagg tgggtctgaa acgataatgg
     4681 tgaatatccc tgcctaactc tattcactat agaaagtaca gcaaaaacta ttcttaaacc
     4741 taccaagcct cctactatca ttatgaataa ttttatatac cacagccaat ttgttatgtt
     4801 aaaccaattc cacaaacttg cccatttatc taattccaat aattcttgtt cattcttttc
     4861 ttgctggttt tgcgattctt caattaagga gtgtattaag cttgtgtaat tgttaatttc
     4921 tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt tccagagatt tattactcca
     4981 actagcattc caaggcacag cagtggtgca aatgagtttt ccagagcaac cccaaatccc
     5041 caggagctgt tgatccttta ggtatctttc cacagccagg attcttgcct ggagctgctt
     5101 gatgccccag actgtgagtt gcaacagatg ctgttgcgcc tcaatagccc tcagcaaatt
     5161 gttctgctgc tgcactatac cagacaataa ttgtctggcc tgtaccgtca gcgtcattga
     5221 cgctgcgccc atagtgcttc ctgctgctcc caagaaccca aggaacaaag ctcctattcc
     5281 cactgctctt ttttctctct gcaccactct tctctttgcc ttggtgggtg ctactcctaa
     5341 tggttcaatt tttactactt tatatttata taattcactt ctccaattgt ccctcatatc
     5401 tcctccttat ggtgttttac taaacttttc catgttctaa tcctcatcct gtctacttgc
     5461 cacacaatca tcacctgcca tctgttttcc ataatcccta atgatctttg cttttcttct
     5521 tggcactact tttatgtcac tattatcttg tattactact gccccttcac ctttccagag
     5581 gagctttgct ggtcctttcc aaagtggatt tctgctgtcc ctgtaataaa cccgaaaatt
     5641 ttgaattttt gtaatttgtt tttgtaattc tttagtttgt atgtctgttg ctattatgtc
     5701 tactattctt tcccctgcac tgtacccccc aatcccccct tttcttttaa aattgtggat
     5761 gaatactgcc atttgtactg ctgtcttaag atgttcagcc tgatctctta cctgtcctat
     5821 aattttcttt aattctttat tcatagattc tactactcct tgactttggg gattgtaggg
     5881 aatttccagg tctgaagatc ctgctgcttg ctgtgctttt ttcttacttt tgttttgctc
     5941 ttcctctatc ttgtctaaag cttccttggt gtcttttatc tctatccttt gatgcacaca
     6001 atagagggtt gctactgtat tatataatga tctaagttct tctgatcctg tctgaaggga
     6061 tggttgtagc tgtcccagta tttgtctaca gccttctgat gtttctaaca ggccagtatt
     6121 aactgcgaat cgttctagct ccctgcttgc ccatactata tgttttaatt tatatttttt
     6181 ctttccccct ggccttaacc gaattttttc ccatcgcgat ctaattctcc cccgcttaat
     6241 actgacgctc tcgcacccat ctctctcctt ctagcctccg ctagtcaaaa tttttggcgt
     6301 actcaccagt cgccgcccct cgcctcttgc cgtgcgcgct tcagcaagcc gagtcctgcg
     6361 tcgagagagc tcctctggtt tccctttcgc tttcaagtcc ctgttcgggc gccactgcta
     6421 gagattttcc acactgacta aaagggtctg agggatctct agttaccaga gtcacacaac
     6481 agacgggcac acactacttg aagcactcaa ggcaagcttt attgaggctt aagcagtggg
     6541 ttccctagtt agccagagag ctcccaggct cagatctggt ctaaccagag agacccagta
     6601 caggcaaaaa gcagctgctt atatgcagga tctgagggct cgccactccc cagtcccgcc
     6661 caggccacgc ctcccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga
     6721 aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg
     6781 tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac
     6841 cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca
     6901 tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt
     6961 accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt
     7021 gattactatt aataactagt caataatcaa tgtcgaccca ggtggcactt ttcggggaaa
     7081 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     7141 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     7201 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     7261 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     7321 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     7381 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     7441 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     7501 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     7561 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     7621 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     7681 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     7741 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     7801 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     7861 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     7921 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     7981 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     8041 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     8101 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     8161 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     8221 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     8281 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     8341 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     8401 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     8461 tgccagtggc gataagtcgt gtcttaccgg
//
