LOCUS       sequence-407494-        8253 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, TSG101_YWHAB from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8253)
  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 8253)
  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_TSG101_YWHAB
FEATURES             Location/Qualifiers
     source          1..8253
                     /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(3014..3724,3725..3727,3728..3730))
                     /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(3056..3077)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3405..3424
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3664..3685
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3727..3736
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3812..4015)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3841..3861)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4016..4319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4576..4620)
                     /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    4805..5038
                     /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    5398..5515
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6007..6132
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6179..6436
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6438..6817
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6848..6952
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6953..7021,7022..7813)
                     /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(7171..7190)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7984..8253,1..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 ccggttcagt
     1801 tttctccctc acctgcatct ccttcatcac cctgattctc tgacgtccac agtgtaagat
     1861 tatcgcgaag aagttgcata ataagtgtgc tatccttata tgattcttcg ttcagtgtat
     1921 ccaattcggc gatagcttca tcaaaagctg tcttcgccaa actacacgct ttctctggac
     1981 tatttaagat ctcataataa aacactgaaa agttaagcgc caagccaaga cggattgggt
     2041 gggttggctg catctccttt ttagaaatct cgaaggcttc ttggtaagcc tgctgggaat
     2101 tgctcacagt cgtctgctta ttgtcacccg aagcgacttc agacaaatag cggaagtaat
     2161 cccctttcat ttttaagtag aatacttttg attcaggttg tgtggcattc gggatcaggt
     2221 acttgtccag caactcaagt acgtcattgc aaatatcctg caattcggcc tcgatcttct
     2281 cgcggtactc ttttcccatt tgctgctttt tttcattacg ttctgtttcg actgttgtaa
     2341 ccggtggttg ggaggggtat tgtgaggatg tggttgcagg gtatggccca ccgggagggt
     2401 aagggcaacc aggatagcca gaagggttag gagggtaacc tgacgggtat gggctaatgc
     2461 cgcccggcat accgggcatg tatgacgtat ttggcggccc ggtagcctga tacggcgggt
     2521 aggaagctga aatggggcgg ctaaacaccg ggggttcgtc accgaacacg acgatcatca
     2581 cctggatcag acctaacagg tccgattgag ggtgtttcca ttcgtgaaga tagggtaagt
     2641 agatcttgcc attggcatca acgtgttttc cagttttgat cgtcattgag gaagtgggtt
     2701 tgacgaagca aatgggcggg ttatacggat aagtatccaa taaccaaagg caaatcggaa
     2761 tgttgtaggt gtttccacgg tatggaacgg gaattgtgcc ggtaaggttc attaactcgc
     2821 gagaagaacc gtcattaaag acatagctat ccagcacagg tttcagatct ttatacaaag
     2881 tgatcacgtt gactgtttca cgaaccgtca aatcgcgata cttgtacttg gacaccattt
     2941 ttttcagttg tgattcgctc actgccattc cccgggaagc ggccgcactt cccccggagc
     3001 ctccgcttcc gcccttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga
     3061 actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagc ttggactggg
     3121 tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct
     3181 ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc ttgaagttca
     3241 ccttgatgcc gttcttctgc ttgtcggcca tgatatagac gttgtggctg ttgtagttgt
     3301 actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga
     3361 tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt
     3421 cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga
     3481 agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg
     3541 tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg
     3601 tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt
     3661 ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct
     3721 tgctcaccat ggtggcgacc ggtggatccc gggcccgcgg taccgtcgac tgcagaattc
     3781 ggttcactaa acgagcccgg ttcactaaac gagctctgct tatatagacc tcccaccgta
     3841 cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc
     3901 gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc
     3961 cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg
     4021 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     4081 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     4141 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     4201 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     4261 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     4321 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat taataactag
     4381 cacgcgttcg aagcttgagc tcggtcccct cggggttggg aggtgggtct gaaacgataa
     4441 tggtgaatat ccctgcctaa ctctattcac tatagaaagt acagcaaaaa ctattcttaa
     4501 acctaccaag cctcctacta tcattatgaa taattttata taccacagcc aatttgttat
     4561 gttaaaccaa ttccacaaac ttgcccattt atctaattcc aataattctt gttcattctt
     4621 ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt aagcttgtgt aattgttaat
     4681 ttctctgtcc cactccatcc aggtcgtgtg attccaaatc tgttccagag atttattact
     4741 ccaactagca ttccaaggca cagcagtggt gcaaatgagt tttccagagc aaccccaaat
     4801 ccccaggagc tgttgatcct ttaggtatct ttccacagcc aggattcttg cctggagctg
     4861 cttgatgccc cagactgtga gttgcaacag atgctgttgc gcctcaatag ccctcagcaa
     4921 attgttctgc tgctgcacta taccagacaa taattgtctg gcctgtaccg tcagcgtcat
     4981 tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac ccaaggaaca aagctcctat
     5041 tcccactgct cttttttctc tctgcaccac tcttctcttt gccttggtgg gtgctactcc
     5101 taatggttca atttttacta ctttatattt atataattca cttctccaat tgtccctcat
     5161 atctcctcct tatggtgttt tactaaactt ttccatgttc taatcctcat cctgtctact
     5221 tgccacacaa tcatcacctg ccatctgttt tccataatcc ctaatgatct ttgcttttct
     5281 tcttggcact acttttatgt cactattatc ttgtattact actgcccctt cacctttcca
     5341 gaggagcttt gctggtcctt tccaaagtgg atttctgctg tccctgtaat aaacccgaaa
     5401 attttgaatt tttgtaattt gtttttgtaa ttctttagtt tgtatgtctg ttgctattat
     5461 gtctactatt ctttcccctg cactgtaccc cccaatcccc ccttttcttt taaaattgtg
     5521 gatgaatact gccatttgta ctgctgtctt aagatgttca gcctgatctc ttacctgtcc
     5581 tataattttc tttaattctt tattcataga ttctactact ccttgacttt ggggattgta
     5641 gggaatttcc aggtctgaag atcctgctgc ttgctgtgct tttttcttac ttttgttttg
     5701 ctcttcctct atcttgtcta aagcttcctt ggtgtctttt atctctatcc tttgatgcac
     5761 acaatagagg gttgctactg tattatataa tgatctaagt tcttctgatc ctgtctgaag
     5821 ggatggttgt agctgtccca gtatttgtct acagccttct gatgtttcta acaggccagt
     5881 attaactgcg aatcgttcta gctccctgct tgcccatact atatgtttta atttatattt
     5941 tttctttccc cctggcctta accgaatttt ttcccatcgc gatctaattc tcccccgctt
     6001 aatactgacg ctctcgcacc catctctctc cttctagcct ccgctagtca aaatttttgg
     6061 cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc gcttcagcaa gccgagtcct
     6121 gcgtcgagag agctcctctg gtttcccttt cgctttcaag tccctgttcg ggcgccactg
     6181 ctagagattt tccacactga ctaaaagggt ctgagggatc tctagttacc agagtcacac
     6241 aacagacggg cacacactac ttgaagcact caaggcaagc tttattgagg cttaagcagt
     6301 gggttcccta gttagccaga gagctcccag gctcagatct ggtctaacca gagagaccca
     6361 gtacaggcaa aaagcagctg cttatatgca ggatctgagg gctcgccact ccccagtccc
     6421 gcccaggcca cgcctcccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
     6481 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
     6541 acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt
     6601 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga
     6661 acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca
     6721 tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta
     6781 attgattact attaataact agtcaataat caatgtcgac ccaggtggca cttttcgggg
     6841 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     6901 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     6961 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     7021 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     7081 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     7141 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     7201 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     7261 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     7321 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     7381 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     7441 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     7501 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     7561 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     7621 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     7681 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     7741 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     7801 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     7861 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     7921 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     7981 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     8041 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     8101 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     8161 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     8221 tgctgccagt ggcgataagt cgtgtcttac cgg
//
