LOCUS       sequence-407495-        8688 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, UNK_B4GALNT2 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8688)
  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 8688)
  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_UNK_B4GALNT2
FEATURES             Location/Qualifiers
     source          1..8688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     22..41
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(260..364)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        395..774
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             776..1033
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1080..1205
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1697..1814
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    2174..2407
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2592..2636
                     /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"
     enhancer        2893..3196
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3197..3400
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3351..3371
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3476..3485
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3482..3484,3485..3487,3488..4198)
                     /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(3527..3548)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3788..3807)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4135..4156
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6100..6119)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6165..6220
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6219..6238)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6581..6597)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6581..6597)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6594..6616)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6605..6621
                     /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)."
     promoter        complement(join(6629..6635,6636..6653,6654..6659))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6674..6695
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6754..6949
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6800..6935
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6955..7089
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6992..7011)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7046..7065
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7157..7174)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7328..7916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7408..7427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8087..8688,1..190,191..259))
                     /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"
ORIGIN
        1 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
       61 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
      121 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
      181 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
      241 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
      301 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
      361 cgcgcacatt tccccgaaaa gtgccacctg ggtcgacatt gattattgac tagttattaa
      421 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      481 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      541 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      601 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      661 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      721 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catgggaggc
      781 gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa gcagctgctt
      841 tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
      901 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
      961 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     1021 aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc
     1081 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
     1141 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
     1201 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
     1261 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
     1321 tcgcagttaa tactggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
     1381 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
     1441 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
     1501 tagaggaaga gcaaaacaaa agtaagaaaa aagcacagca agcagcagga tcttcagacc
     1561 tggaaattcc ctacaatccc caaagtcaag gagtagtaga atctatgaat aaagaattaa
     1621 agaaaattat aggacaggta agagatcagg ctgaacatct taagacagca gtacaaatgg
     1681 cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa
     1741 gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa
     1801 aaattcaaaa ttttcgggtt tattacaggg acagcagaaa tccactttgg aaaggaccag
     1861 caaagctcct ctggaaaggt gaaggggcag tagtaataca agataatagt gacataaaag
     1921 tagtgccaag aagaaaagca aagatcatta gggattatgg aaaacagatg gcaggtgatg
     1981 attgtgtggc aagtagacag gatgaggatt agaacatgga aaagtttagt aaaacaccat
     2041 aaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa
     2101 ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa
     2161 gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg
     2221 gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc
     2281 agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct
     2341 ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac
     2401 agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga
     2461 atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt
     2521 gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa
     2581 accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga
     2641 attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag
     2701 gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg
     2761 gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccg agctcaagct
     2821 tcgaacgcgt gctagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     2881 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     2941 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     3001 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     3061 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     3121 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     3181 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     3241 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     3301 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     3361 cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga accgggctcg
     3421 tttagtgaac cgaattctgc agtcgacggt accgcgggcc cgggatccac cggtcgccac
     3481 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     3541 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     3601 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     3661 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     3721 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     3781 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     3841 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     3901 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     3961 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     4021 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     4081 ctacctgagc acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
     4141 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg
     4201 cggaagcgga ggctccgggg gaagtgcggc cgcttcccgg atgagtaagg gtcctggacc
     4261 aggaggatca gcggctagta gtgccccgcc cgccgcgact gctcaagtcc tgcaagctca
     4321 accagagaag cctcaacatt atacatcgcg ctttttgtgg ttgctgaaaa ttttagtcat
     4381 tattttagtt ctgggtattg tcgggttcat gtttggtagt atgtttttgc aagcggtgtt
     4441 ctctagtccg aaacccgagc tgccgagtcc ggcacccggt gttcagaaat tgaagctgtt
     4501 accagaggaa cgcttgcgta accttttcag ttacgatggc atctggcttt ttccgaaaaa
     4561 tcagtgtaaa tgtgaagcca acaaagagca agggggctat aattttcagg acgcatacgg
     4621 tcaatcggat cttccagcag taaaagcgcg tcgtcaggca gaatttgagc actttcagcg
     4681 ccgtgagggc ttgccacgtc cgttacccct tctggtacaa cccaacctgc cgtttggcta
     4741 ccctgtacat ggcgtggagg ttatgccatt acacaccgtg ccgatcccag gccttcagtt
     4801 cgagggacct gatgccccgg tttacgaagt gactttaacc gcttctcttg ggacattaaa
     4861 caccttagca gacgtaccgg actccgtggt tcagggtcgt ggccagaagc agcttatcat
     4921 tagcacttca gaccgtaaac tgttaaaatt catcctgcag catgtaacct atacatcaac
     4981 gggttaccaa caccagaaag tcgacatcgt gtcactggaa tcgcgctcaa gcgtggctaa
     5041 attcccggtc accatccgcc atccagttat ccccaaatta tacgaccctg gccctgagcg
     5101 taagctgcgt aacttagtca caattgcgac gaaaacattt ctgcgccctc acaaattaat
     5161 gattatgtta cgttcgattc gtgagtacta cccggacttg acggtcatcg tcgccgacga
     5221 cagccaaaag ccgcttgaga ttaaagataa ccatgtagaa tactatacaa tgcccttcgg
     5281 taagggttgg ttcgcgggtc gcaatttagc aatcagtcaa gttacaacga aatatgtatt
     5341 atgggttgat gatgattttt tattcaacga ggaaacaaaa attgaggtct tagtagacgt
     5401 tttagagaag actgagcttg atgtagtcgg cggttcagta ttgggaaacg tgtttcagtt
     5461 caaattgttg ttggagcaaa gcgaaaatgg cgcttgctta cacaagcgta tggggttttt
     5521 tcagccactg gacggtttcc cctcctgtgt tgtcaccagt ggtgttgtaa actttttcct
     5581 tgctcacacc gagcgccttc aacgcgtcgg gttcgatccc cgcttacagc gcgtcgccca
     5641 ctccgagttc tttatcgacg ggctgggtac cttgttggta ggatcgtgcc ctgaggtaat
     5701 catcggacat caaagtcgca gccctgtagt agatagtgag cttgctgcac tggaaaagac
     5761 gtacaataca tatcgctcga acacgcttac ccgcgtccaa ttcaaactgg cgcttcacta
     5821 tttcaagaac catttgcagt gtgcggcttg aaccggttct agataagatt acggtacctt
     5881 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     5941 gactggaagg gctaattcac tcccaaagaa gacaagatct gctttttgcc tgtactgggt
     6001 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     6061 ttaagcctca ataaagcttc agctgctcga gctagcagat ctttttccct ctgccaaaaa
     6121 ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat
     6181 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     6241 ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca
     6301 tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc
     6361 ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat
     6421 attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact
     6481 agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg
     6541 gagatccctc gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     6601 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6661 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6721 tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt
     6781 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     6841 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     6901 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg
     6961 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     7021 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     7081 gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     7141 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     7201 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7261 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7321 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7381 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7441 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7501 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7561 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7621 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7681 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7741 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     7801 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7861 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7921 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7981 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     8041 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     8101 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     8161 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     8221 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     8281 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     8341 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     8401 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     8461 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8521 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8581 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8641 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgct
//
