LOCUS       sequence-408146-        7506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, MSMP_RGP1 from patient-derived 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7506)
  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 7506)
  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_MSMP_RGP1
FEATURES             Location/Qualifiers
     source          1..7506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(113..217)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        248..627
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             629..886
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    933..1058
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1550..1667
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    2027..2260
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2445..2489
                     /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        2746..3049
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3050..3253
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3204..3224
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3329..3338
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3335..3337,3338..3340,3341..4051)
                     /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(3380..3401)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3641..3660)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3988..4009
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4771..4790)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4836..4891
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4890..4909)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5252..5268)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5252..5268)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5265..5287)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5276..5292
                     /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(5300..5306,5307..5324,5325..5330))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5345..5366
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5425..5620
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5471..5606
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5626..5760
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5663..5682)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5717..5736
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5828..5845)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5999..6587)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6079..6098)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6758..7506,1..43,44..112))
                     /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     7381..7400
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
       61 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
      121 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
      181 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
      241 ctgggtcgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      301 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      361 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      421 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      481 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      541 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      601 tacgtattag tcatcgctat taccatggga ggcgtggcct gggcgggact ggggagtggc
      661 gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta
      721 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
      781 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
      841 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
      901 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
      961 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     1021 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     1081 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     1141 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatactggc ctgttagaaa
     1201 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     1261 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     1321 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     1381 aaaaagcaca gcaagcagca ggatcttcag acctggaaat tccctacaat ccccaaagtc
     1441 aaggagtagt agaatctatg aataaagaat taaagaaaat tataggacag gtaagagatc
     1501 aggctgaaca tcttaagaca gcagtacaaa tggcagtatt catccacaat tttaaaagaa
     1561 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
     1621 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
     1681 gggacagcag aaatccactt tggaaaggac cagcaaagct cctctggaaa ggtgaagggg
     1741 cagtagtaat acaagataat agtgacataa aagtagtgcc aagaagaaaa gcaaagatca
     1801 ttagggatta tggaaaacag atggcaggtg atgattgtgt ggcaagtaga caggatgagg
     1861 attagaacat ggaaaagttt agtaaaacac cataaggagg agatatgagg gacaattgga
     1921 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     1981 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     2041 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     2101 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     2161 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     2221 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     2281 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     2341 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     2401 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     2461 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     2521 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     2581 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     2641 acctcccaac cccgagggga ccgagctcaa gcttcgaacg cgtgctagtt attaatagta
     2701 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     2761 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     2821 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     2881 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     2941 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     3001 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     3061 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     3121 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     3181 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     3241 tataagcaga gctcgtttag tgaaccgggc tcgtttagtg aaccgaattc tgcagtcgac
     3301 ggtaccgcgg gcccgggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt
     3361 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     3421 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     3481 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     3541 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     3601 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     3661 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     3721 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     3781 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     3841 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     3901 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag
     3961 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     4021 gatcactctc ggcatggacg agctgtacaa gggcggaagc ggaggctccg ggggaagtgc
     4081 ggccgcttcc cggggaatgg cccttcgcat gttatgggcg ggccaagcca aaggcatttt
     4141 aggcggatgg ggtattatct gcttggtaat gagcttgttg ttgcaacatc cgggggttta
     4201 tagtaagtgc tactttcagg cgcaggcccc ctgtcattat gaaggcaagt acttcaccct
     4261 tggggagagc tggctgcgta aagactgctt tcattgcaca tgccttcatc cagtcggcgt
     4321 aggatgctgc gacacttccc aacatcctat cgacttcccc gccggttgtg aggttcgtca
     4381 agaagcaggg acctgtcagt tttctttggt acagaaaagt gatacgttct cgtgggatct
     4441 gccgatcaaa gtcctgccaa cttcacccac attggcgagt tatgccgcgc ctggtccgag
     4501 tacttctact atcacgatct gaaccggttc tagataagat tacggtacct ttaagaccaa
     4561 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     4621 ggctaattca ctcccaaaga agacaagatc tgctttttgc ctgtactggg tctctctggt
     4681 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     4741 aataaagctt cagctgctcg agctagcaga tctttttccc tctgccaaaa attatgggga
     4801 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
     4861 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     4921 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc
     4981 tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt
     5041 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt
     5101 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt
     5161 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct
     5221 cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5281 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5341 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5401 gaaacctgtc gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct
     5461 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     5521 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     5581 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca
     5641 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5701 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     5761 cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5821 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5881 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5941 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6001 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6061 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6121 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6181 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6241 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6301 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6361 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6421 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6481 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6541 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6601 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6661 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6721 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6781 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6841 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6901 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6961 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7021 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7081 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7141 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7201 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7261 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7321 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7381 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7441 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7501 caccca
//
