LOCUS       sequence-488145-        7689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_122, derived from human fusion protein NUP98_KDM5A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7689)
  AUTHORS   Maiti S, Tripathi S, Baggett DW, Phillips AH, Park CG, Wang J, 
            Wahiduzzaman, Freyaldenhoven WT, Kinger S, Pioso BJ, Bollinger JC, 
            Somjee R, Lang B, Babu MM, Kriwacki RW
  TITLE     Proteome-wide computational analyses reveal links between protein 
            condensate formation and RNA biology.
  JOURNAL   Sci Adv. 2025 Dec 5;11(49):eady1420. doi: 10.1126/sciadv.ady1420. 
            Epub 2025 Dec 3.
  PUBMED    41337585
REFERENCE   2  (bases 1 to 7689)
  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: "Sci Adv."; 
            date: "2025-12-5"; volume: "11(49):eady1420. doi"; pages: " 
            10.1126/sciadv.ady1420. Epub 2025 Dec 3"
COMMENT     Sequence Label: CL20_mEGFP_FOidr_122
FEATURES             Location/Qualifiers
     source          1..7689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     181..200
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    199..254
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     300..319
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(1222..1932,1933..1935,1936..1938))
                     /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(1264..1285)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1613..1632
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1872..1893
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1935..1944
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2020..2223)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2049..2069)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2224..2527
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2784..2828)
                     /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    3013..3246
                     /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    3606..3723
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4215..4340
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4387..4644
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4646..5025
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5056..5160
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5161..5229,5230..6021)
                     /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(5379..5398)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6192..6780
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6681..6700
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6934..6951
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7019..7153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7043..7062)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7097..7116
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7159..7354)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7173..7308
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7413..7434
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7449..7454,7455..7472,7473..7479)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7487..7503
                     /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     7492..7514
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7511..7527
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7511..7527
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aaaaaaaatc taacctcaag tcaaggcttt tctatggaat aaggaatgga cagcaggggg
       61 ctgtttcata tactgatgac ctctttatag ccaacctttg ttcatggcag ccagcatatg
      121 ggcatatgtt gccaaactct aaaccaaata ctcattctga tgttttaaat gatttgccct
      181 cccatatgtc cttccgagtg agagacacaa aaaattccaa cacactattg caatgaaaat
      241 aaatttcctt tattagccag aagtcagatg ctcaaggggc ttcatgatgt ccccataatt
      301 tttggcagag ggaaaaagat ctgctagctc gagcagctga agctttattg aggcttaagc
      361 agtgggttcc ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac
      421 ccagtacagg caaaaagcag atcttgtctt ctttgggagt gaattagccc ttccagtccc
      481 cccttttctt ttaaaaagtg gctaagatct acagctgcct tgtaagtcat tggtcttaaa
      541 ggtaccgtaa tcttatctag aaccggttca ctctgctcct gaccagtcat actttgaggg
      601 gatgtccagc acctttttct ctcttttctt ttcagtgctc tctttcacaa gttcaacttt
      661 ggctgcagca gccttctcct tctttttctt tctctcttct tcttttgcta gtttcttggc
      721 cagtttattc agctcctttg atttgtctgc acctaatttt aatttcttct ttctaggttt
      781 gtccattttc tttaactcct tggacttctg ttttccttct ccaaaaagtt gctctacctt
      841 ttctagcttc cgtttccgtt tcttctctga agagtccttt cctttcactt ttagtggttt
      901 ctcttccatg ctgtcatcct cttccttctt cttagggtct gacatcggat tccggaagag
      961 aggagagtct ccaaaaggtg agtatgttag actattgatg tgctgctgga gaacagcctg
     1021 ctgggcagca gaagcatttg gatctgtcaa agctactggg gcctgggggg ctccaaagcc
     1081 caaagtggct gtcgtagtat taaatccagg ggccccaaag gctcctgtac caagaggccc
     1141 tccaatctta ggttggttgt tcccaaacaa agatgcctgc cgggaagcgg ccgcacttcc
     1201 cccggagcct ccgcttccgc ccttgtacag ctcgtccatg ccgagagtga tcccggcggc
     1261 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt
     1321 ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
     1381 gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt
     1441 gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt
     1501 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
     1561 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
     1621 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
     1681 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta
     1741 ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa
     1801 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     1861 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     1921 ctcgcccttg ctcaccatgg tggcgaccgg tggatcccgg gcccgcggta ccgtcgactg
     1981 cagaattcgg ttcactaaac gagcccggtt cactaaacga gctctgctta tatagacctc
     2041 ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg
     2101 aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact
     2161 tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat
     2221 caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa
     2281 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg
     2341 cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc
     2401 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat
     2461 tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta
     2521 tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg attactatta
     2581 ataactagca cgcgttcgaa gcttgagctc ggtcccctcg gggttgggag gtgggtctga
     2641 aacgataatg gtgaatatcc ctgcctaact ctattcacta tagaaagtac agcaaaaact
     2701 attcttaaac ctaccaagcc tcctactatc attatgaata attttatata ccacagccaa
     2761 tttgttatgt taaaccaatt ccacaaactt gcccatttat ctaattccaa taattcttgt
     2821 tcattctttt cttgctggtt ttgcgattct tcaattaagg agtgtattaa gcttgtgtaa
     2881 ttgttaattt ctctgtccca ctccatccag gtcgtgtgat tccaaatctg ttccagagat
     2941 ttattactcc aactagcatt ccaaggcaca gcagtggtgc aaatgagttt tccagagcaa
     3001 ccccaaatcc ccaggagctg ttgatccttt aggtatcttt ccacagccag gattcttgcc
     3061 tggagctgct tgatgcccca gactgtgagt tgcaacagat gctgttgcgc ctcaatagcc
     3121 ctcagcaaat tgttctgctg ctgcactata ccagacaata attgtctggc ctgtaccgtc
     3181 agcgtcattg acgctgcgcc catagtgctt cctgctgctc ccaagaaccc aaggaacaaa
     3241 gctcctattc ccactgctct tttttctctc tgcaccactc ttctctttgc cttggtgggt
     3301 gctactccta atggttcaat ttttactact ttatatttat ataattcact tctccaattg
     3361 tccctcatat ctcctcctta tggtgtttta ctaaactttt ccatgttcta atcctcatcc
     3421 tgtctacttg ccacacaatc atcacctgcc atctgttttc cataatccct aatgatcttt
     3481 gcttttcttc ttggcactac ttttatgtca ctattatctt gtattactac tgccccttca
     3541 cctttccaga ggagctttgc tggtcctttc caaagtggat ttctgctgtc cctgtaataa
     3601 acccgaaaat tttgaatttt tgtaatttgt ttttgtaatt ctttagtttg tatgtctgtt
     3661 gctattatgt ctactattct ttcccctgca ctgtaccccc caatcccccc ttttctttta
     3721 aaattgtgga tgaatactgc catttgtact gctgtcttaa gatgttcagc ctgatctctt
     3781 acctgtccta taattttctt taattcttta ttcatagatt ctactactcc ttgactttgg
     3841 ggattgtagg gaatttccag gtctgaagat cctgctgctt gctgtgcttt tttcttactt
     3901 ttgttttgct cttcctctat cttgtctaaa gcttccttgg tgtcttttat ctctatcctt
     3961 tgatgcacac aatagagggt tgctactgta ttatataatg atctaagttc ttctgatcct
     4021 gtctgaaggg atggttgtag ctgtcccagt atttgtctac agccttctga tgtttctaac
     4081 aggccagtat taactgcgaa tcgttctagc tccctgcttg cccatactat atgttttaat
     4141 ttatattttt tctttccccc tggccttaac cgaatttttt cccatcgcga tctaattctc
     4201 ccccgcttaa tactgacgct ctcgcaccca tctctctcct tctagcctcc gctagtcaaa
     4261 atttttggcg tactcaccag tcgccgcccc tcgcctcttg ccgtgcgcgc ttcagcaagc
     4321 cgagtcctgc gtcgagagag ctcctctggt ttccctttcg ctttcaagtc cctgttcggg
     4381 cgccactgct agagattttc cacactgact aaaagggtct gagggatctc tagttaccag
     4441 agtcacacaa cagacgggca cacactactt gaagcactca aggcaagctt tattgaggct
     4501 taagcagtgg gttccctagt tagccagaga gctcccaggc tcagatctgg tctaaccaga
     4561 gagacccagt acaggcaaaa agcagctgct tatatgcagg atctgagggc tcgccactcc
     4621 ccagtcccgc ccaggccacg cctcccatgg taatagcgat gactaatacg tagatgtact
     4681 gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac
     4741 cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt
     4801 gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta
     4861 ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag
     4921 gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg
     4981 accccgtaat tgattactat taataactag tcaataatca atgtcgaccc aggtggcact
     5041 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5101 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5161 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5221 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5281 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5341 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5401 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5461 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5521 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5581 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5641 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5701 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5761 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5821 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5881 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5941 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6001 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6061 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6121 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6181 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6241 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6301 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6361 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6421 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6481 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6541 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6601 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6661 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6721 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6781 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6841 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     6901 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     6961 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagcgga
     7021 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     7081 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     7141 caataaacaa gttagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
     7201 ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca
     7261 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
     7321 gggcgggact atggttgctg actaattgag atgcggatcc gctggcacga caggtttccc
     7381 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     7441 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     7501 caatttcaca caggaaacag ctatgaccat gattacgcca agcttgggct gcaggtcgag
     7561 ggatctccat aagagaagag ggacagctat gactgggagt agtcaggaga ggaggaaaaa
     7621 tctggctagt aaaacatgta aggaaaattt tagggatgtt aaagaaaaaa ataacacaaa
     7681 acaaaatat
//
