LOCUS       sequence-408178-        7836 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, ZC3H4_POLR2I from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7836)
  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 7836)
  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_ZC3H4_POLR2I
FEATURES             Location/Qualifiers
     source          1..7836
                     /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(2597..3307,3308..3310,3311..3313))
                     /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(2639..2660)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2988..3007
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3247..3268
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3310..3319
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3395..3598)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3424..3444)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3599..3902
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4159..4203)
                     /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    4388..4621
                     /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    4981..5098
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5590..5715
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5762..6019
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6021..6400
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6431..6535
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6536..6604,6605..7396)
                     /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(6754..6773)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7567..7836,1..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 ccggttcact
     1801 cggtccatct gtgcccacag tggggcgccg tgcaaacata atagagtctc atagcgtctt
     1861 cggcgcgagc ggagtgagat tgaaaaaaaa ctgcctcttt gtggccacac ttttgacaag
     1921 ggtgatcctc tgtgcgaggg agcgtcggat cttgtgaaac atctgcaatt atctgcgtga
     1981 gctcatcaac ctcgtgcgtg atcttattca cataaatgca tgaattatcg gcctcttgtt
     2041 gatagtcaca gtttcgacag ggcgcgtagg gtggtgagta ctctctgtac ttccggtggc
     2101 ccttctcact aggactgaag tctgaatcgt cagagaagtc ggagaagtca tcgcttgaag
     2161 aggcgtgcct tttatgttta cttttcctgc gttttttgga ccgcctcttc tctttttccc
     2221 gctccttctt gcgttttctc ttgaggcgcc tatgggactt ttcttcatcg gagtcactat
     2281 gatgtttttc tcctttctcc ttccgagatc tctccggtcc cccggacgtg tcttgcgtct
     2341 cctctgctcc gtcatcttca agttcgcctt cctccaactc accgtcttct ctatcgtctg
     2401 gcaatggaag tctatgatgc agaaggtgtg gagttgctgg ccttgcatca gggctacaag
     2461 gtggcgggct aggggtgctt ggtggtggtg gagatggagg aggcggtgac tcactaggcg
     2521 ggggtggtgg ggttccaggc gcggcttcca ttccccggga agcggccgca cttcccccgg
     2581 agcctccgct tccgcccttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     2641 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agcttggact
     2701 gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     2761 gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt
     2821 tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt
     2881 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     2941 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
     3001 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     3061 agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca
     3121 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca
     3181 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
     3241 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
     3301 ccttgctcac catggtggcg accggtggat cccgggcccg cggtaccgtc gactgcagaa
     3361 ttcggttcac taaacgagcc cggttcacta aacgagctct gcttatatag acctcccacc
     3421 gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt
     3481 cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa
     3541 atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca
     3601 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
     3661 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
     3721 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
     3781 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
     3841 tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa
     3901 cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac tattaataac
     3961 tagcacgcgt tcgaagcttg agctcggtcc cctcggggtt gggaggtggg tctgaaacga
     4021 taatggtgaa tatccctgcc taactctatt cactatagaa agtacagcaa aaactattct
     4081 taaacctacc aagcctccta ctatcattat gaataatttt atataccaca gccaatttgt
     4141 tatgttaaac caattccaca aacttgccca tttatctaat tccaataatt cttgttcatt
     4201 cttttcttgc tggttttgcg attcttcaat taaggagtgt attaagcttg tgtaattgtt
     4261 aatttctctg tcccactcca tccaggtcgt gtgattccaa atctgttcca gagatttatt
     4321 actccaacta gcattccaag gcacagcagt ggtgcaaatg agttttccag agcaacccca
     4381 aatccccagg agctgttgat cctttaggta tctttccaca gccaggattc ttgcctggag
     4441 ctgcttgatg ccccagactg tgagttgcaa cagatgctgt tgcgcctcaa tagccctcag
     4501 caaattgttc tgctgctgca ctataccaga caataattgt ctggcctgta ccgtcagcgt
     4561 cattgacgct gcgcccatag tgcttcctgc tgctcccaag aacccaagga acaaagctcc
     4621 tattcccact gctctttttt ctctctgcac cactcttctc tttgccttgg tgggtgctac
     4681 tcctaatggt tcaattttta ctactttata tttatataat tcacttctcc aattgtccct
     4741 catatctcct ccttatggtg ttttactaaa cttttccatg ttctaatcct catcctgtct
     4801 acttgccaca caatcatcac ctgccatctg ttttccataa tccctaatga tctttgcttt
     4861 tcttcttggc actactttta tgtcactatt atcttgtatt actactgccc cttcaccttt
     4921 ccagaggagc tttgctggtc ctttccaaag tggatttctg ctgtccctgt aataaacccg
     4981 aaaattttga atttttgtaa tttgtttttg taattcttta gtttgtatgt ctgttgctat
     5041 tatgtctact attctttccc ctgcactgta ccccccaatc cccccttttc ttttaaaatt
     5101 gtggatgaat actgccattt gtactgctgt cttaagatgt tcagcctgat ctcttacctg
     5161 tcctataatt ttctttaatt ctttattcat agattctact actccttgac tttggggatt
     5221 gtagggaatt tccaggtctg aagatcctgc tgcttgctgt gcttttttct tacttttgtt
     5281 ttgctcttcc tctatcttgt ctaaagcttc cttggtgtct tttatctcta tcctttgatg
     5341 cacacaatag agggttgcta ctgtattata taatgatcta agttcttctg atcctgtctg
     5401 aagggatggt tgtagctgtc ccagtatttg tctacagcct tctgatgttt ctaacaggcc
     5461 agtattaact gcgaatcgtt ctagctccct gcttgcccat actatatgtt ttaatttata
     5521 ttttttcttt ccccctggcc ttaaccgaat tttttcccat cgcgatctaa ttctcccccg
     5581 cttaatactg acgctctcgc acccatctct ctccttctag cctccgctag tcaaaatttt
     5641 tggcgtactc accagtcgcc gcccctcgcc tcttgccgtg cgcgcttcag caagccgagt
     5701 cctgcgtcga gagagctcct ctggtttccc tttcgctttc aagtccctgt tcgggcgcca
     5761 ctgctagaga ttttccacac tgactaaaag ggtctgaggg atctctagtt accagagtca
     5821 cacaacagac gggcacacac tacttgaagc actcaaggca agctttattg aggcttaagc
     5881 agtgggttcc ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac
     5941 ccagtacagg caaaaagcag ctgcttatat gcaggatctg agggctcgcc actccccagt
     6001 cccgcccagg ccacgcctcc catggtaata gcgatgacta atacgtagat gtactgccaa
     6061 gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca
     6121 ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca
     6181 gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg
     6241 ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg
     6301 ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc
     6361 gtaattgatt actattaata actagtcaat aatcaatgtc gacccaggtg gcacttttcg
     6421 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     6481 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     6541 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     6601 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     6661 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     6721 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     6781 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     6841 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     6901 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     6961 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     7021 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     7081 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     7141 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     7201 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     7261 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     7321 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     7381 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     7441 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7501 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7561 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     7621 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7681 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     7741 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7801 ggctgctgcc agtggcgata agtcgtgtct taccgg
//
