LOCUS       sequence-488217-        7533 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_200, derived from human fusion protein TRIOBP_RBFOX2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7533)
  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 7533)
  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_200
FEATURES             Location/Qualifiers
     source          1..7533
                     /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(1066..1776,1777..1779,1780..1782))
                     /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(1108..1129)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1457..1476
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1716..1737
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1779..1788
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1864..2067)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1893..1913)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2068..2371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2628..2672)
                     /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    2857..3090
                     /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    3450..3567
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4059..4184
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4231..4488
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4490..4869
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4900..5004
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5005..5073,5074..5865)
                     /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(5223..5242)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6036..6624
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6525..6544
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6778..6795
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6863..6997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6887..6906)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6941..6960
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7003..7198)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7017..7152
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7257..7278
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7293..7298,7299..7316,7317..7323)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7331..7347
                     /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     7336..7358
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7355..7371
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7355..7371
                     /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 aaaacgaaac gggatgttac taacgtgaag
      601 acgcttagga gtcgatttag actcgctgtt ttcactactc tgtgtttggg attgctgccc
      661 gtcagtttgg gctccaccct cggtcgtaag cgacccgttc tgtgttgacg gggaattgct
      721 actctgttca ccgtgagcct gggttgaccc gtacaaggta agattgtgtt ctccagtttg
      781 gcccgcataa tcctgagtgt ggggcacacc gtactccgtc gggatgccgt tttggggcgg
      841 agggggaaag ggaatagtcg tgaagggttg gaccatagca tcaggggtcg tagtcggctc
      901 ctgattgcct tggtagcggg ctccgtgagc ctcctccgga tgaaagcagt tttgacaacg
      961 gttttgagtc agaatgttcg cctcgaaatg ctcacataac gcgtctcctg ggacttcttc
     1021 catccgggaa gcggccgcac ttcccccgga gcctccgctt ccgcccttgt acagctcgtc
     1081 catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt
     1141 ctcgttgggg tctttgctca gcttggactg ggtgctcagg tagtggttgt cgggcagcag
     1201 cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc
     1261 gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct gcttgtcggc
     1321 catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc
     1381 gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa
     1441 cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg
     1501 gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta
     1561 gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc agggcacggg
     1621 cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc
     1681 gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat
     1741 gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga ccggtggatc
     1801 ccgggcccgc ggtaccgtcg actgcagaat tcggttcact aaacgagccc ggttcactaa
     1861 acgagctctg cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg
     1921 ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc
     1981 attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc
     2041 attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta
     2101 ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt
     2161 accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa
     2221 gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt
     2281 tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc
     2341 aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa
     2401 tgaccccgta attgattact attaataact agcacgcgtt cgaagcttga gctcggtccc
     2461 ctcggggttg ggaggtgggt ctgaaacgat aatggtgaat atccctgcct aactctattc
     2521 actatagaaa gtacagcaaa aactattctt aaacctacca agcctcctac tatcattatg
     2581 aataatttta tataccacag ccaatttgtt atgttaaacc aattccacaa acttgcccat
     2641 ttatctaatt ccaataattc ttgttcattc ttttcttgct ggttttgcga ttcttcaatt
     2701 aaggagtgta ttaagcttgt gtaattgtta atttctctgt cccactccat ccaggtcgtg
     2761 tgattccaaa tctgttccag agatttatta ctccaactag cattccaagg cacagcagtg
     2821 gtgcaaatga gttttccaga gcaaccccaa atccccagga gctgttgatc ctttaggtat
     2881 ctttccacag ccaggattct tgcctggagc tgcttgatgc cccagactgt gagttgcaac
     2941 agatgctgtt gcgcctcaat agccctcagc aaattgttct gctgctgcac tataccagac
     3001 aataattgtc tggcctgtac cgtcagcgtc attgacgctg cgcccatagt gcttcctgct
     3061 gctcccaaga acccaaggaa caaagctcct attcccactg ctcttttttc tctctgcacc
     3121 actcttctct ttgccttggt gggtgctact cctaatggtt caatttttac tactttatat
     3181 ttatataatt cacttctcca attgtccctc atatctcctc cttatggtgt tttactaaac
     3241 ttttccatgt tctaatcctc atcctgtcta cttgccacac aatcatcacc tgccatctgt
     3301 tttccataat ccctaatgat ctttgctttt cttcttggca ctacttttat gtcactatta
     3361 tcttgtatta ctactgcccc ttcacctttc cagaggagct ttgctggtcc tttccaaagt
     3421 ggatttctgc tgtccctgta ataaacccga aaattttgaa tttttgtaat ttgtttttgt
     3481 aattctttag tttgtatgtc tgttgctatt atgtctacta ttctttcccc tgcactgtac
     3541 cccccaatcc ccccttttct tttaaaattg tggatgaata ctgccatttg tactgctgtc
     3601 ttaagatgtt cagcctgatc tcttacctgt cctataattt tctttaattc tttattcata
     3661 gattctacta ctccttgact ttggggattg tagggaattt ccaggtctga agatcctgct
     3721 gcttgctgtg cttttttctt acttttgttt tgctcttcct ctatcttgtc taaagcttcc
     3781 ttggtgtctt ttatctctat cctttgatgc acacaataga gggttgctac tgtattatat
     3841 aatgatctaa gttcttctga tcctgtctga agggatggtt gtagctgtcc cagtatttgt
     3901 ctacagcctt ctgatgtttc taacaggcca gtattaactg cgaatcgttc tagctccctg
     3961 cttgcccata ctatatgttt taatttatat tttttctttc cccctggcct taaccgaatt
     4021 ttttcccatc gcgatctaat tctcccccgc ttaatactga cgctctcgca cccatctctc
     4081 tccttctagc ctccgctagt caaaattttt ggcgtactca ccagtcgccg cccctcgcct
     4141 cttgccgtgc gcgcttcagc aagccgagtc ctgcgtcgag agagctcctc tggtttccct
     4201 ttcgctttca agtccctgtt cgggcgccac tgctagagat tttccacact gactaaaagg
     4261 gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact acttgaagca
     4321 ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca gagagctccc
     4381 aggctcagat ctggtctaac cagagagacc cagtacaggc aaaaagcagc tgcttatatg
     4441 caggatctga gggctcgcca ctccccagtc ccgcccaggc cacgcctccc atggtaatag
     4501 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
     4561 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
     4621 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     4681 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc
     4741 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact
     4801 ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata
     4861 atcaatgtcg acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4921 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4981 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5041 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5101 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5161 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5221 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5281 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5341 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5401 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5461 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5521 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5581 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5641 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5701 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5761 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5821 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5881 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5941 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6001 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6061 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6121 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6181 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6241 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6301 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6361 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6421 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6481 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6541 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6601 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6661 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6721 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     6781 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     6841 tggccgattc attaatgcag cggatccaga catgataaga tacattgatg agtttggaca
     6901 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     6961 tttatttgta accattataa gctgcaataa acaagttagc tttttgcaaa agcctaggcc
     7021 tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc
     7081 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
     7141 taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcgg
     7201 atccgctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     7261 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     7321 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     7381 gccaagcttg ggctgcaggt cgagggatct ccataagaga agagggacag ctatgactgg
     7441 gagtagtcag gagaggagga aaaatctggc tagtaaaaca tgtaaggaaa attttaggga
     7501 tgttaaagaa aaaaataaca caaaacaaaa tat
//
