LOCUS       sequence-488126-        7497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_101, derived from human fusion protein MBNL1_MEIS2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7497)
  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 7497)
  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_101
FEATURES             Location/Qualifiers
     source          1..7497
                     /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(1030..1740,1741..1743,1744..1746))
                     /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(1072..1093)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1421..1440
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1680..1701
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1743..1752
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1828..2031)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1857..1877)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2032..2335
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2592..2636)
                     /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    2821..3054
                     /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    3414..3531
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4023..4148
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4195..4452
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4454..4833
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4864..4968
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4969..5037,5038..5829)
                     /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(5187..5206)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6000..6588
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6489..6508
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6742..6759
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6827..6961
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6851..6870)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6905..6924
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6967..7162)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6981..7116
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7221..7242
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7257..7262,7263..7280,7281..7287)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7295..7311
                     /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     7300..7322
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7319..7335
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7319..7335
                     /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 ctgtgcgtga atatccataa cttgaccgcc
      601 cacatttgga tcaacgctat tcagcattgt tggactttga gcggacatgg tcattccggg
      661 atgagttgga ggtcctccat gcatcatcat tgcgggatga tgtggatggc tcggcaagta
      721 tgaatgcatc ggcggcccat ggcgtaattg cgtggggtgt ggcgtcattt ggggaggggt
      781 gtacgacggc tgagccattg acatccccat gggaccgccc tgactaacat aatcgccggg
      841 catactctga agaccggcag gacggattcc catgtgttgt tgcccgtcca aaacaaacga
      901 ccccattggt tgcccctcag gggagtaggc ggcgccttgg gatacggcgc gattactctg
      961 gtcaatcatt ggctgtacga tgcggcgccg ggaagcggcc gcacttcccc cggagcctcc
     1021 gcttccgccc ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc
     1081 cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg actgggtgct
     1141 caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta
     1201 gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt
     1261 gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc
     1321 cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg
     1381 gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc
     1441 cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc
     1501 gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt
     1561 cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag
     1621 cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac
     1681 gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct
     1741 caccatggtg gcgaccggtg gatcccgggc ccgcggtacc gtcgactgca gaattcggtt
     1801 cactaaacga gcccggttca ctaaacgagc tctgcttata tagacctccc accgtacacg
     1861 cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg
     1921 attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg
     1981 tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat
     2041 agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg
     2101 ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat
     2161 atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg
     2221 tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg
     2281 gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa
     2341 ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagcacg
     2401 cgttcgaagc ttgagctcgg tcccctcggg gttgggaggt gggtctgaaa cgataatggt
     2461 gaatatccct gcctaactct attcactata gaaagtacag caaaaactat tcttaaacct
     2521 accaagcctc ctactatcat tatgaataat tttatatacc acagccaatt tgttatgtta
     2581 aaccaattcc acaaacttgc ccatttatct aattccaata attcttgttc attcttttct
     2641 tgctggtttt gcgattcttc aattaaggag tgtattaagc ttgtgtaatt gttaatttct
     2701 ctgtcccact ccatccaggt cgtgtgattc caaatctgtt ccagagattt attactccaa
     2761 ctagcattcc aaggcacagc agtggtgcaa atgagttttc cagagcaacc ccaaatcccc
     2821 aggagctgtt gatcctttag gtatctttcc acagccagga ttcttgcctg gagctgcttg
     2881 atgccccaga ctgtgagttg caacagatgc tgttgcgcct caatagccct cagcaaattg
     2941 ttctgctgct gcactatacc agacaataat tgtctggcct gtaccgtcag cgtcattgac
     3001 gctgcgccca tagtgcttcc tgctgctccc aagaacccaa ggaacaaagc tcctattccc
     3061 actgctcttt tttctctctg caccactctt ctctttgcct tggtgggtgc tactcctaat
     3121 ggttcaattt ttactacttt atatttatat aattcacttc tccaattgtc cctcatatct
     3181 cctccttatg gtgttttact aaacttttcc atgttctaat cctcatcctg tctacttgcc
     3241 acacaatcat cacctgccat ctgttttcca taatccctaa tgatctttgc ttttcttctt
     3301 ggcactactt ttatgtcact attatcttgt attactactg ccccttcacc tttccagagg
     3361 agctttgctg gtcctttcca aagtggattt ctgctgtccc tgtaataaac ccgaaaattt
     3421 tgaatttttg taatttgttt ttgtaattct ttagtttgta tgtctgttgc tattatgtct
     3481 actattcttt cccctgcact gtacccccca atcccccctt ttcttttaaa attgtggatg
     3541 aatactgcca tttgtactgc tgtcttaaga tgttcagcct gatctcttac ctgtcctata
     3601 attttcttta attctttatt catagattct actactcctt gactttgggg attgtaggga
     3661 atttccaggt ctgaagatcc tgctgcttgc tgtgcttttt tcttactttt gttttgctct
     3721 tcctctatct tgtctaaagc ttccttggtg tcttttatct ctatcctttg atgcacacaa
     3781 tagagggttg ctactgtatt atataatgat ctaagttctt ctgatcctgt ctgaagggat
     3841 ggttgtagct gtcccagtat ttgtctacag ccttctgatg tttctaacag gccagtatta
     3901 actgcgaatc gttctagctc cctgcttgcc catactatat gttttaattt atattttttc
     3961 tttccccctg gccttaaccg aattttttcc catcgcgatc taattctccc ccgcttaata
     4021 ctgacgctct cgcacccatc tctctccttc tagcctccgc tagtcaaaat ttttggcgta
     4081 ctcaccagtc gccgcccctc gcctcttgcc gtgcgcgctt cagcaagccg agtcctgcgt
     4141 cgagagagct cctctggttt ccctttcgct ttcaagtccc tgttcgggcg ccactgctag
     4201 agattttcca cactgactaa aagggtctga gggatctcta gttaccagag tcacacaaca
     4261 gacgggcaca cactacttga agcactcaag gcaagcttta ttgaggctta agcagtgggt
     4321 tccctagtta gccagagagc tcccaggctc agatctggtc taaccagaga gacccagtac
     4381 aggcaaaaag cagctgctta tatgcaggat ctgagggctc gccactcccc agtcccgccc
     4441 aggccacgcc tcccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     4501 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     4561 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     4621 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     4681 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     4741 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     4801 attactatta ataactagtc aataatcaat gtcgacccag gtggcacttt tcggggaaat
     4861 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4921 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4981 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5041 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5101 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5161 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5221 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5281 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5341 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5401 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5461 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5521 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5581 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5641 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5701 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5761 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5821 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5881 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5941 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6001 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6061 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6121 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6181 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6241 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6301 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6361 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6421 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6481 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6541 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6601 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6661 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     6721 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     6781 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagcggatc cagacatgat
     6841 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     6901 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     6961 tagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     7021 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     7081 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     7141 ggttgctgac taattgagat gcggatccgc tggcacgaca ggtttcccga ctggaaagcg
     7201 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     7261 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     7321 ggaaacagct atgaccatga ttacgccaag cttgggctgc aggtcgaggg atctccataa
     7381 gagaagaggg acagctatga ctgggagtag tcaggagagg aggaaaaatc tggctagtaa
     7441 aacatgtaag gaaaatttta gggatgttaa agaaaaaaat aacacaaaac aaaatat
//
