LOCUS       sequence-487965-        7629 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_052, derived from human fusion protein DDX3X_MRAS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7629)
  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 7629)
  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_052
FEATURES             Location/Qualifiers
     source          1..7629
                     /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(1162..1872,1873..1875,1876..1878))
                     /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(1204..1225)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1553..1572
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1812..1833
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1875..1884
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1960..2163)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1989..2009)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2164..2467
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2724..2768)
                     /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    2953..3186
                     /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    3546..3663
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4155..4280
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4327..4584
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4586..4965
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4996..5100
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5101..5169,5170..5961)
                     /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(5319..5338)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6132..6720
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6621..6640
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6874..6891
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6959..7093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6983..7002)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7037..7056
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7099..7294)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7113..7248
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7353..7374
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7389..7394,7395..7412,7413..7419)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7427..7443
                     /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     7432..7454
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7451..7467
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7451..7467
                     /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 tgagaaactt tcaatgtggg gcgggcaatt
      601 gttccccgtt gcctcgacgg gaatgtcgtc atacttttca aaattaatcc cggtgttgcc
      661 tcccgaaaac agctcttgct ctaagcgttc gctaggagga agtggcttac tccaatcgtc
      721 ctcgtctgat ttgtcgcacc aacgagagtt gccgccgcgc tcgaacttac cgaatcccga
      781 gcggtctccg cgggagccaa tgccgtcgta gtcggagcgt ccacgatcgt cgaaacgtcc
      841 acgtgagcca gatccgcgat cagaaaagaa gctcgacttc ccgcgtgagt ctgagcggga
      901 cccgaaggaa gagtatgcat ccttatcctt cgaactcgac cagccggagc tatccttatc
      961 gtaaaagccc ttcgttgcct cacggttgcg cagatgaggc ggaatgtagc ggcctttgct
     1021 cgcggtggaa cctccacttt gattatcact agaattaagg tccagcccag caaattgctg
     1081 gtctaaccct aacgcatttt ccaccgcaac gtgagacatc cgggaagcgg ccgcacttcc
     1141 cccggagcct ccgcttccgc ccttgtacag ctcgtccatg ccgagagtga tcccggcggc
     1201 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt
     1261 ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
     1321 gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt
     1381 gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt
     1441 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
     1501 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
     1561 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
     1621 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta
     1681 ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa
     1741 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     1801 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     1861 ctcgcccttg ctcaccatgg tggcgaccgg tggatcccgg gcccgcggta ccgtcgactg
     1921 cagaattcgg ttcactaaac gagcccggtt cactaaacga gctctgctta tatagacctc
     1981 ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg
     2041 aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact
     2101 tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat
     2161 caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa
     2221 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg
     2281 cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc
     2341 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat
     2401 tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta
     2461 tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg attactatta
     2521 ataactagca cgcgttcgaa gcttgagctc ggtcccctcg gggttgggag gtgggtctga
     2581 aacgataatg gtgaatatcc ctgcctaact ctattcacta tagaaagtac agcaaaaact
     2641 attcttaaac ctaccaagcc tcctactatc attatgaata attttatata ccacagccaa
     2701 tttgttatgt taaaccaatt ccacaaactt gcccatttat ctaattccaa taattcttgt
     2761 tcattctttt cttgctggtt ttgcgattct tcaattaagg agtgtattaa gcttgtgtaa
     2821 ttgttaattt ctctgtccca ctccatccag gtcgtgtgat tccaaatctg ttccagagat
     2881 ttattactcc aactagcatt ccaaggcaca gcagtggtgc aaatgagttt tccagagcaa
     2941 ccccaaatcc ccaggagctg ttgatccttt aggtatcttt ccacagccag gattcttgcc
     3001 tggagctgct tgatgcccca gactgtgagt tgcaacagat gctgttgcgc ctcaatagcc
     3061 ctcagcaaat tgttctgctg ctgcactata ccagacaata attgtctggc ctgtaccgtc
     3121 agcgtcattg acgctgcgcc catagtgctt cctgctgctc ccaagaaccc aaggaacaaa
     3181 gctcctattc ccactgctct tttttctctc tgcaccactc ttctctttgc cttggtgggt
     3241 gctactccta atggttcaat ttttactact ttatatttat ataattcact tctccaattg
     3301 tccctcatat ctcctcctta tggtgtttta ctaaactttt ccatgttcta atcctcatcc
     3361 tgtctacttg ccacacaatc atcacctgcc atctgttttc cataatccct aatgatcttt
     3421 gcttttcttc ttggcactac ttttatgtca ctattatctt gtattactac tgccccttca
     3481 cctttccaga ggagctttgc tggtcctttc caaagtggat ttctgctgtc cctgtaataa
     3541 acccgaaaat tttgaatttt tgtaatttgt ttttgtaatt ctttagtttg tatgtctgtt
     3601 gctattatgt ctactattct ttcccctgca ctgtaccccc caatcccccc ttttctttta
     3661 aaattgtgga tgaatactgc catttgtact gctgtcttaa gatgttcagc ctgatctctt
     3721 acctgtccta taattttctt taattcttta ttcatagatt ctactactcc ttgactttgg
     3781 ggattgtagg gaatttccag gtctgaagat cctgctgctt gctgtgcttt tttcttactt
     3841 ttgttttgct cttcctctat cttgtctaaa gcttccttgg tgtcttttat ctctatcctt
     3901 tgatgcacac aatagagggt tgctactgta ttatataatg atctaagttc ttctgatcct
     3961 gtctgaaggg atggttgtag ctgtcccagt atttgtctac agccttctga tgtttctaac
     4021 aggccagtat taactgcgaa tcgttctagc tccctgcttg cccatactat atgttttaat
     4081 ttatattttt tctttccccc tggccttaac cgaatttttt cccatcgcga tctaattctc
     4141 ccccgcttaa tactgacgct ctcgcaccca tctctctcct tctagcctcc gctagtcaaa
     4201 atttttggcg tactcaccag tcgccgcccc tcgcctcttg ccgtgcgcgc ttcagcaagc
     4261 cgagtcctgc gtcgagagag ctcctctggt ttccctttcg ctttcaagtc cctgttcggg
     4321 cgccactgct agagattttc cacactgact aaaagggtct gagggatctc tagttaccag
     4381 agtcacacaa cagacgggca cacactactt gaagcactca aggcaagctt tattgaggct
     4441 taagcagtgg gttccctagt tagccagaga gctcccaggc tcagatctgg tctaaccaga
     4501 gagacccagt acaggcaaaa agcagctgct tatatgcagg atctgagggc tcgccactcc
     4561 ccagtcccgc ccaggccacg cctcccatgg taatagcgat gactaatacg tagatgtact
     4621 gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac
     4681 cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt
     4741 gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta
     4801 ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag
     4861 gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg
     4921 accccgtaat tgattactat taataactag tcaataatca atgtcgaccc aggtggcact
     4981 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5041 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5101 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5161 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5221 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5281 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5341 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5401 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5461 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5521 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5581 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5641 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5701 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5761 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5821 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5881 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5941 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6001 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6061 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6121 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6181 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6241 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6301 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6361 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6421 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6481 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6541 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6601 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6661 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6721 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6781 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     6841 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     6901 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagcgga
     6961 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     7021 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     7081 caataaacaa gttagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
     7141 ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca
     7201 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
     7261 gggcgggact atggttgctg actaattgag atgcggatcc gctggcacga caggtttccc
     7321 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     7381 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     7441 caatttcaca caggaaacag ctatgaccat gattacgcca agcttgggct gcaggtcgag
     7501 ggatctccat aagagaagag ggacagctat gactgggagt agtcaggaga ggaggaaaaa
     7561 tctggctagt aaaacatgta aggaaaattt tagggatgtt aaagaaaaaa ataacacaaa
     7621 acaaaatat
//
