LOCUS       sequence-408153-        7485 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, PICK1_POLR2F from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7485)
  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 7485)
  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_PICK1_POLR2F
FEATURES             Location/Qualifiers
     source          1..7485
                     /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(2246..2956,2957..2959,2960..2962))
                     /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(2288..2309)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2637..2656
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2896..2917
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2959..2968
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3044..3247)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3073..3093)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3248..3551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3808..3852)
                     /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    4037..4270
                     /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    4630..4747
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5239..5364
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5411..5668
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5670..6049
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6080..6184
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6185..6253,6254..7045)
                     /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(6403..6422)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7216..7485,1..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 ctgtgataat gagctcatct actccccaat cctcataact accatccggc aggtatcttc
     1861 taatgataat cggtattttt cgcgctttga gttccttcat ggcaatcaac agcggatcag
     1921 tctccccttc cagctctacc atcactggcg cacacatagc aatttgcaac gcccgtgtac
     1981 cgaggactcg cgctcgttca tacttcgtca tatacggagt tgtaatcctt ttctggttag
     2041 cttggggtct ttctccactc gggaggatct ctacattttc ctggccttct tcttctgcat
     2101 tctcaaggtc atccaagccc tcgtcttcct ccacatcgtc gaagtcgtcg ccatcaaaga
     2161 gtttatcctc ctctatgtcg taatccaaat cagcgaacat tccccgggaa gcggccgcac
     2221 ttcccccgga gcctccgctt ccgcccttgt acagctcgtc catgccgaga gtgatcccgg
     2281 cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca
     2341 gcttggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg
     2401 gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga
     2461 tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag acgttgtggc
     2521 tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc
     2581 ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct
     2641 tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg
     2701 cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag cactgcacgc
     2761 cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga
     2821 tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga
     2881 acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca
     2941 gctcctcgcc cttgctcacc atggtggcga ccggtggatc ccgggcccgc ggtaccgtcg
     3001 actgcagaat tcggttcact aaacgagccc ggttcactaa acgagctctg cttatataga
     3061 cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt
     3121 ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga
     3181 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc
     3241 gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     3301 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     3361 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     3421 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     3481 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     3541 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
     3601 attaataact agcacgcgtt cgaagcttga gctcggtccc ctcggggttg ggaggtgggt
     3661 ctgaaacgat aatggtgaat atccctgcct aactctattc actatagaaa gtacagcaaa
     3721 aactattctt aaacctacca agcctcctac tatcattatg aataatttta tataccacag
     3781 ccaatttgtt atgttaaacc aattccacaa acttgcccat ttatctaatt ccaataattc
     3841 ttgttcattc ttttcttgct ggttttgcga ttcttcaatt aaggagtgta ttaagcttgt
     3901 gtaattgtta atttctctgt cccactccat ccaggtcgtg tgattccaaa tctgttccag
     3961 agatttatta ctccaactag cattccaagg cacagcagtg gtgcaaatga gttttccaga
     4021 gcaaccccaa atccccagga gctgttgatc ctttaggtat ctttccacag ccaggattct
     4081 tgcctggagc tgcttgatgc cccagactgt gagttgcaac agatgctgtt gcgcctcaat
     4141 agccctcagc aaattgttct gctgctgcac tataccagac aataattgtc tggcctgtac
     4201 cgtcagcgtc attgacgctg cgcccatagt gcttcctgct gctcccaaga acccaaggaa
     4261 caaagctcct attcccactg ctcttttttc tctctgcacc actcttctct ttgccttggt
     4321 gggtgctact cctaatggtt caatttttac tactttatat ttatataatt cacttctcca
     4381 attgtccctc atatctcctc cttatggtgt tttactaaac ttttccatgt tctaatcctc
     4441 atcctgtcta cttgccacac aatcatcacc tgccatctgt tttccataat ccctaatgat
     4501 ctttgctttt cttcttggca ctacttttat gtcactatta tcttgtatta ctactgcccc
     4561 ttcacctttc cagaggagct ttgctggtcc tttccaaagt ggatttctgc tgtccctgta
     4621 ataaacccga aaattttgaa tttttgtaat ttgtttttgt aattctttag tttgtatgtc
     4681 tgttgctatt atgtctacta ttctttcccc tgcactgtac cccccaatcc ccccttttct
     4741 tttaaaattg tggatgaata ctgccatttg tactgctgtc ttaagatgtt cagcctgatc
     4801 tcttacctgt cctataattt tctttaattc tttattcata gattctacta ctccttgact
     4861 ttggggattg tagggaattt ccaggtctga agatcctgct gcttgctgtg cttttttctt
     4921 acttttgttt tgctcttcct ctatcttgtc taaagcttcc ttggtgtctt ttatctctat
     4981 cctttgatgc acacaataga gggttgctac tgtattatat aatgatctaa gttcttctga
     5041 tcctgtctga agggatggtt gtagctgtcc cagtatttgt ctacagcctt ctgatgtttc
     5101 taacaggcca gtattaactg cgaatcgttc tagctccctg cttgcccata ctatatgttt
     5161 taatttatat tttttctttc cccctggcct taaccgaatt ttttcccatc gcgatctaat
     5221 tctcccccgc ttaatactga cgctctcgca cccatctctc tccttctagc ctccgctagt
     5281 caaaattttt ggcgtactca ccagtcgccg cccctcgcct cttgccgtgc gcgcttcagc
     5341 aagccgagtc ctgcgtcgag agagctcctc tggtttccct ttcgctttca agtccctgtt
     5401 cgggcgccac tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta
     5461 ccagagtcac acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga
     5521 ggcttaagca gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac
     5581 cagagagacc cagtacaggc aaaaagcagc tgcttatatg caggatctga gggctcgcca
     5641 ctccccagtc ccgcccaggc cacgcctccc atggtaatag cgatgactaa tacgtagatg
     5701 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
     5761 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
     5821 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
     5881 gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag
     5941 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
     6001 aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtcg acccaggtgg
     6061 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     6121 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     6181 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     6241 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     6301 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     6361 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     6421 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     6481 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     6541 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6601 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6661 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6721 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     6781 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     6841 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6901 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6961 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     7021 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     7081 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     7141 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     7201 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     7261 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     7321 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     7381 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     7441 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgg
//
