LOCUS       sequence-407468-        8028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, RPS6KB1_VMP1 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8028)
  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 8028)
  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_RPS6KB1_VMP1
FEATURES             Location/Qualifiers
     source          1..8028
                     /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(2789..3499,3500..3502,3503..3505))
                     /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(2831..2852)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3180..3199
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3439..3460
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3502..3511
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3587..3790)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3616..3636)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3791..4094
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4351..4395)
                     /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    4580..4813
                     /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    5173..5290
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5782..5907
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5954..6211
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6213..6592
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6623..6727
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6728..6796,6797..7588)
                     /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(6946..6965)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7759..8028,1..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 ccggttcatt
     1801 tagttttttc ctcggagtta aggcgttgtt ggatgcgttt cgcatagctc tgagccatag
     1861 agttaataat gctaaggata aaataacata ccatgacgac aactaatttt tcaaacatcc
     1921 agctcagcca attctcacct tgtggggtac ccatctcgga cttgtgatgc agtttttggc
     1981 gctgagcttc taagtactcc tgaaatggtt tttgcagact gggaccaatt cccggaacag
     2041 ctccgataaa agcaaccatc tgttcaacaa tatgttttga gaatgtaata atcacaaaaa
     2101 ttttctgaat atgcattttg ataattgctt tgccaatcaa cgtagcgcca aagaacgtcc
     2161 agaatggaac caggaagtgt ccgcaggtaa tgcctgccaa gtcaaacaac ggatttggga
     2221 tggacgcaca agccaggatg ccgaaaaacc caactttttg tactaatttc tgaacagcaa
     2281 gtttagcgcg tgaagcaaaa tcttgcgcgc tttcagcgtg ttctaacatc tcttcaaatt
     2341 cttggtattc ttcatcatcg ggctctgcac cactcaggcg ggcagcgcgg gccataaagt
     2401 acgggggcaa ttcaccgatg gcagttccga ttccccacat gcacgcctca atacgcactt
     2461 tggaaatgat ggaccacaaa ctaattgtgc cttcagtgcc ttcctcgtct ggacaaatga
     2521 tctggtcagg atacggcggc tcggggaaat tgactgagtt gcactcgtat gcggccagag
     2581 taacagatgc aatatggggt ccaccttcct ccaactcatc ttctgatccc gcatcttcag
     2641 gctgatccag atcaatatca aatactccag ccatatcctc ggcctcacgg tcacggaaat
     2701 ctggagccgg gtaaaacccg tcacggcggc gacggcgacg cattccccgg gaagcggccg
     2761 cacttccccc ggagcctccg cttccgccct tgtacagctc gtccatgccg agagtgatcc
     2821 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
     2881 tcagcttgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
     2941 tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc
     3001 ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt
     3061 ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga
     3121 tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg
     3181 tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca
     3241 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca
     3301 cgccgtaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc
     3361 agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc
     3421 tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga
     3481 acagctcctc gcccttgctc accatggtgg cgaccggtgg atcccgggcc cgcggtaccg
     3541 tcgactgcag aattcggttc actaaacgag cccggttcac taaacgagct ctgcttatat
     3601 agacctccca ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac
     3661 attttggaaa gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt
     3721 ggagacttgg aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa
     3781 accgcatcac catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt
     3841 cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa
     3901 tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta
     3961 aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg
     4021 tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg
     4081 taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt
     4141 actattaata actagcacgc gttcgaagct tgagctcggt cccctcgggg ttgggaggtg
     4201 ggtctgaaac gataatggtg aatatccctg cctaactcta ttcactatag aaagtacagc
     4261 aaaaactatt cttaaaccta ccaagcctcc tactatcatt atgaataatt ttatatacca
     4321 cagccaattt gttatgttaa accaattcca caaacttgcc catttatcta attccaataa
     4381 ttcttgttca ttcttttctt gctggttttg cgattcttca attaaggagt gtattaagct
     4441 tgtgtaattg ttaatttctc tgtcccactc catccaggtc gtgtgattcc aaatctgttc
     4501 cagagattta ttactccaac tagcattcca aggcacagca gtggtgcaaa tgagttttcc
     4561 agagcaaccc caaatcccca ggagctgttg atcctttagg tatctttcca cagccaggat
     4621 tcttgcctgg agctgcttga tgccccagac tgtgagttgc aacagatgct gttgcgcctc
     4681 aatagccctc agcaaattgt tctgctgctg cactatacca gacaataatt gtctggcctg
     4741 taccgtcagc gtcattgacg ctgcgcccat agtgcttcct gctgctccca agaacccaag
     4801 gaacaaagct cctattccca ctgctctttt ttctctctgc accactcttc tctttgcctt
     4861 ggtgggtgct actcctaatg gttcaatttt tactacttta tatttatata attcacttct
     4921 ccaattgtcc ctcatatctc ctccttatgg tgttttacta aacttttcca tgttctaatc
     4981 ctcatcctgt ctacttgcca cacaatcatc acctgccatc tgttttccat aatccctaat
     5041 gatctttgct tttcttcttg gcactacttt tatgtcacta ttatcttgta ttactactgc
     5101 cccttcacct ttccagagga gctttgctgg tcctttccaa agtggatttc tgctgtccct
     5161 gtaataaacc cgaaaatttt gaatttttgt aatttgtttt tgtaattctt tagtttgtat
     5221 gtctgttgct attatgtcta ctattctttc ccctgcactg taccccccaa tccccccttt
     5281 tcttttaaaa ttgtggatga atactgccat ttgtactgct gtcttaagat gttcagcctg
     5341 atctcttacc tgtcctataa ttttctttaa ttctttattc atagattcta ctactccttg
     5401 actttgggga ttgtagggaa tttccaggtc tgaagatcct gctgcttgct gtgctttttt
     5461 cttacttttg ttttgctctt cctctatctt gtctaaagct tccttggtgt cttttatctc
     5521 tatcctttga tgcacacaat agagggttgc tactgtatta tataatgatc taagttcttc
     5581 tgatcctgtc tgaagggatg gttgtagctg tcccagtatt tgtctacagc cttctgatgt
     5641 ttctaacagg ccagtattaa ctgcgaatcg ttctagctcc ctgcttgccc atactatatg
     5701 ttttaattta tattttttct ttccccctgg ccttaaccga attttttccc atcgcgatct
     5761 aattctcccc cgcttaatac tgacgctctc gcacccatct ctctccttct agcctccgct
     5821 agtcaaaatt tttggcgtac tcaccagtcg ccgcccctcg cctcttgccg tgcgcgcttc
     5881 agcaagccga gtcctgcgtc gagagagctc ctctggtttc cctttcgctt tcaagtccct
     5941 gttcgggcgc cactgctaga gattttccac actgactaaa agggtctgag ggatctctag
     6001 ttaccagagt cacacaacag acgggcacac actacttgaa gcactcaagg caagctttat
     6061 tgaggcttaa gcagtgggtt ccctagttag ccagagagct cccaggctca gatctggtct
     6121 aaccagagag acccagtaca ggcaaaaagc agctgcttat atgcaggatc tgagggctcg
     6181 ccactcccca gtcccgccca ggccacgcct cccatggtaa tagcgatgac taatacgtag
     6241 atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc
     6301 catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact
     6361 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt
     6421 ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt
     6481 cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga
     6541 actaatgacc ccgtaattga ttactattaa taactagtca ataatcaatg tcgacccagg
     6601 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6661 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6721 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6781 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6841 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6901 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6961 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     7021 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     7081 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     7141 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     7201 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     7261 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     7321 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     7381 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     7441 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     7501 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     7561 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     7621 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     7681 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7741 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7801 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7861 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     7921 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7981 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgg
//
