LOCUS       sequence-407486-        8628 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, TMPRSS2_ERG from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8628)
  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 8628)
  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_TMPRSS2_ERG
FEATURES             Location/Qualifiers
     source          1..8628
                     /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(3389..4099,4100..4102,4103..4105))
                     /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(3431..3452)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3780..3799
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4039..4060
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4102..4111
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4187..4390)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4216..4236)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4391..4694
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4951..4995)
                     /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    5180..5413
                     /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    5773..5890
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6382..6507
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6554..6811
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6813..7192
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7223..7327
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7328..7396,7397..8188)
                     /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(7546..7565)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8359..8628,1..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 ccggttcaat
     1801 agtaggtgcc caagtgagac ggcatgtgac tggttggcaa gcgcgtgtta gggtagatgc
     1861 cccccgtggg tgagttccaa tatggattag gtgcagcaaa gaagctagag ctagtcactg
     1921 gtaaagccgg gggatgcggt gccacaaagt tcatcttctg tggatgagcg tgatacgatc
     1981 ccatgtacgg caagtctgat ggatatttat aaagcgaact ctctggagga tgtggttgta
     2041 acgcttgggc gatcccatga aaatcaaatt tgtaagcgta acgtttgcca tgaacctttg
     2101 tcatgatatt tttatcatag taataacgta acgcgcggga cagtttgtcg tagttcatgt
     2161 tcggcttgga cttacgctca ccccagcggc gtgcaacttc gtctggatcg gtcattttaa
     2221 attcgccatt tgtaccctcc caggtgatgc atgaagaatt actcgaatcg gacagcagtt
     2281 ccagaagaaa ttgccataac tggatctggc ccgagcccgg atttgctaaa cgggagcttg
     2341 tcggacccag gatctgatag gggtccagtt gtggacgttg atcctctgtc ttgggtaccg
     2401 ttgatggact gggctgggca gccttacttt ggggagtggg atgaccgtgt ccggtccacg
     2461 cggagcggcg gggcggttcg tagggcaaat cgggacgagt tgtgatgcgc tgtgttgcct
     2521 cagggtagac acttgtattg gggaaaatga acgctgcccc accggtgttg cgagcgtgca
     2581 tcaggcgtgg gctgttttgt aatgccttgt caacgtcgtc tgaagtcagg tgtggaaggg
     2641 gtgtctcgcg taaataatga agatgtgaca acaggatgtc ggcattgtag gaaggcgtaa
     2701 gacgttgaaa atcgtcttta gtcattttac acagttcctt accgtcaatg ttctggaata
     2761 aaagaatgtt tacgtctggc agaccatact cctttacagc ccattcaagc cactgacgta
     2821 cgtggtcggt agaccaaagc gttgggtcag cgggaacaat cacgcgacgc tcattagttg
     2881 tcatgttcgg tggaggcatg tgtttctcct ccatgtagga cccgtagttc atccctaccg
     2941 tatccggact gccaaccatt ttgcctcctt tggctacgct acactcatcg ggcgaattgc
     3001 gtgacccatt aacctgcgat ggattgcatt ccattttgat agtgacacgt gcggggggct
     3061 gagacaacca gtcttgctgc gggacacgcg gcgacatttt acttgtctgc ccataatccg
     3121 acgacgaaga tgctgtcatt tcggttttcg ctaaatgcgg tgtcccataa gcacactcaa
     3181 ataaggattg atcctcggag actacactca gagcttcgct attcagtgcc atcttactat
     3241 tgtcaacggc atccagcaga gacaaataac ggctgtgttc aatatgacca gctgcaccgc
     3301 gttcctcaca tcctgactcg ccgccaggcg gagcgggagg cattccccgg gaagcggccg
     3361 cacttccccc ggagcctccg cttccgccct tgtacagctc gtccatgccg agagtgatcc
     3421 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
     3481 tcagcttgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
     3541 tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc
     3601 ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt
     3661 ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga
     3721 tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg
     3781 tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca
     3841 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca
     3901 cgccgtaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc
     3961 agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc
     4021 tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga
     4081 acagctcctc gcccttgctc accatggtgg cgaccggtgg atcccgggcc cgcggtaccg
     4141 tcgactgcag aattcggttc actaaacgag cccggttcac taaacgagct ctgcttatat
     4201 agacctccca ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac
     4261 attttggaaa gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt
     4321 ggagacttgg aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa
     4381 accgcatcac catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt
     4441 cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa
     4501 tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta
     4561 aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg
     4621 tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg
     4681 taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt
     4741 actattaata actagcacgc gttcgaagct tgagctcggt cccctcgggg ttgggaggtg
     4801 ggtctgaaac gataatggtg aatatccctg cctaactcta ttcactatag aaagtacagc
     4861 aaaaactatt cttaaaccta ccaagcctcc tactatcatt atgaataatt ttatatacca
     4921 cagccaattt gttatgttaa accaattcca caaacttgcc catttatcta attccaataa
     4981 ttcttgttca ttcttttctt gctggttttg cgattcttca attaaggagt gtattaagct
     5041 tgtgtaattg ttaatttctc tgtcccactc catccaggtc gtgtgattcc aaatctgttc
     5101 cagagattta ttactccaac tagcattcca aggcacagca gtggtgcaaa tgagttttcc
     5161 agagcaaccc caaatcccca ggagctgttg atcctttagg tatctttcca cagccaggat
     5221 tcttgcctgg agctgcttga tgccccagac tgtgagttgc aacagatgct gttgcgcctc
     5281 aatagccctc agcaaattgt tctgctgctg cactatacca gacaataatt gtctggcctg
     5341 taccgtcagc gtcattgacg ctgcgcccat agtgcttcct gctgctccca agaacccaag
     5401 gaacaaagct cctattccca ctgctctttt ttctctctgc accactcttc tctttgcctt
     5461 ggtgggtgct actcctaatg gttcaatttt tactacttta tatttatata attcacttct
     5521 ccaattgtcc ctcatatctc ctccttatgg tgttttacta aacttttcca tgttctaatc
     5581 ctcatcctgt ctacttgcca cacaatcatc acctgccatc tgttttccat aatccctaat
     5641 gatctttgct tttcttcttg gcactacttt tatgtcacta ttatcttgta ttactactgc
     5701 cccttcacct ttccagagga gctttgctgg tcctttccaa agtggatttc tgctgtccct
     5761 gtaataaacc cgaaaatttt gaatttttgt aatttgtttt tgtaattctt tagtttgtat
     5821 gtctgttgct attatgtcta ctattctttc ccctgcactg taccccccaa tccccccttt
     5881 tcttttaaaa ttgtggatga atactgccat ttgtactgct gtcttaagat gttcagcctg
     5941 atctcttacc tgtcctataa ttttctttaa ttctttattc atagattcta ctactccttg
     6001 actttgggga ttgtagggaa tttccaggtc tgaagatcct gctgcttgct gtgctttttt
     6061 cttacttttg ttttgctctt cctctatctt gtctaaagct tccttggtgt cttttatctc
     6121 tatcctttga tgcacacaat agagggttgc tactgtatta tataatgatc taagttcttc
     6181 tgatcctgtc tgaagggatg gttgtagctg tcccagtatt tgtctacagc cttctgatgt
     6241 ttctaacagg ccagtattaa ctgcgaatcg ttctagctcc ctgcttgccc atactatatg
     6301 ttttaattta tattttttct ttccccctgg ccttaaccga attttttccc atcgcgatct
     6361 aattctcccc cgcttaatac tgacgctctc gcacccatct ctctccttct agcctccgct
     6421 agtcaaaatt tttggcgtac tcaccagtcg ccgcccctcg cctcttgccg tgcgcgcttc
     6481 agcaagccga gtcctgcgtc gagagagctc ctctggtttc cctttcgctt tcaagtccct
     6541 gttcgggcgc cactgctaga gattttccac actgactaaa agggtctgag ggatctctag
     6601 ttaccagagt cacacaacag acgggcacac actacttgaa gcactcaagg caagctttat
     6661 tgaggcttaa gcagtgggtt ccctagttag ccagagagct cccaggctca gatctggtct
     6721 aaccagagag acccagtaca ggcaaaaagc agctgcttat atgcaggatc tgagggctcg
     6781 ccactcccca gtcccgccca ggccacgcct cccatggtaa tagcgatgac taatacgtag
     6841 atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc
     6901 catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact
     6961 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt
     7021 ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt
     7081 cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga
     7141 actaatgacc ccgtaattga ttactattaa taactagtca ataatcaatg tcgacccagg
     7201 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     7261 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     7321 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     7381 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     7441 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     7501 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     7561 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     7621 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     7681 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     7741 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     7801 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     7861 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     7921 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     7981 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     8041 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     8101 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     8161 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     8221 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     8281 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     8341 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     8401 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     8461 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     8521 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     8581 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgg
//
