LOCUS       sequence-488124-        7587 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_099, derived from human fusion protein MBNL1_MEIS2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7587)
  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 7587)
  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_099
FEATURES             Location/Qualifiers
     source          1..7587
                     /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(1120..1830,1831..1833,1834..1836))
                     /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(1162..1183)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1511..1530
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1770..1791
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1833..1842
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1918..2121)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1947..1967)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2122..2425
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2682..2726)
                     /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    2911..3144
                     /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    3504..3621
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4113..4238
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4285..4542
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4544..4923
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4954..5058
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5059..5127,5128..5919)
                     /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(5277..5296)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6090..6678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6579..6598
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6832..6849
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6917..7051
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6941..6960)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6995..7014
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7057..7252)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7071..7206
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7311..7332
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7347..7352,7353..7370,7371..7377)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7385..7401
                     /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     7390..7412
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7409..7425
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7409..7425
                     /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 actgcagaca tcgccccccg caacccctgg
      601 ctcacgtggt gtgcaagtag caagttcaca cttttcaaag accaaggcaa gtaaggggaa
      661 aaggggatgg ccgtaaatgg cgtctttgtc gcgcttcagc gcgtcgttta ccgcactccc
      721 catggacgct ggcataacat tgggatgcgg tgcatgcgcc ccgtaatgct gcgtagcgtg
      781 taagggggga ccatggttca agtgatgtac tggtgggatg ggacggggcg cgtgagggtc
      841 gccgtacatg gacgctggaa ctcctacacc atccatccca ccgtagtggg gtaattcgtc
      901 atagactggt tgcaggggcg cacctggcat catggcatta gccagctgca tttgttgtgc
      961 taacatggcc atgttctttt gttgaatcaa attattacgg ccattaattt ctaattgtgt
     1021 tttaagatga ggaggtggat gtaaatattt gcaattttcg cggctacaac gtcccatccg
     1081 ggaagcggcc gcacttcccc cggagcctcc gcttccgccc ttgtacagct cgtccatgcc
     1141 gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt
     1201 ggggtctttg ctcagcttgg actgggtgct caggtagtgg ttgtcgggca gcagcacggg
     1261 gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc tgccgtcctc
     1321 gatgttgtgg cggatcttga agttcacctt gatgccgttc ttctgcttgt cggccatgat
     1381 atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc
     1441 cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac
     1501 ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta
     1561 gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct
     1621 gaagcactgc acgccgtagg tcagggtggt cacgagggtg ggccagggca cgggcagctt
     1681 gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc cctcgccctc
     1741 gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac
     1801 caccccggtg aacagctcct cgcccttgct caccatggtg gcgaccggtg gatcccgggc
     1861 ccgcggtacc gtcgactgca gaattcggtt cactaaacga gcccggttca ctaaacgagc
     1921 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga
     1981 gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac
     2041 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat
     2101 gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca
     2161 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     2221 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     2281 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     2341 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     2401 gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc
     2461 cgtaattgat tactattaat aactagcacg cgttcgaagc ttgagctcgg tcccctcggg
     2521 gttgggaggt gggtctgaaa cgataatggt gaatatccct gcctaactct attcactata
     2581 gaaagtacag caaaaactat tcttaaacct accaagcctc ctactatcat tatgaataat
     2641 tttatatacc acagccaatt tgttatgtta aaccaattcc acaaacttgc ccatttatct
     2701 aattccaata attcttgttc attcttttct tgctggtttt gcgattcttc aattaaggag
     2761 tgtattaagc ttgtgtaatt gttaatttct ctgtcccact ccatccaggt cgtgtgattc
     2821 caaatctgtt ccagagattt attactccaa ctagcattcc aaggcacagc agtggtgcaa
     2881 atgagttttc cagagcaacc ccaaatcccc aggagctgtt gatcctttag gtatctttcc
     2941 acagccagga ttcttgcctg gagctgcttg atgccccaga ctgtgagttg caacagatgc
     3001 tgttgcgcct caatagccct cagcaaattg ttctgctgct gcactatacc agacaataat
     3061 tgtctggcct gtaccgtcag cgtcattgac gctgcgccca tagtgcttcc tgctgctccc
     3121 aagaacccaa ggaacaaagc tcctattccc actgctcttt tttctctctg caccactctt
     3181 ctctttgcct tggtgggtgc tactcctaat ggttcaattt ttactacttt atatttatat
     3241 aattcacttc tccaattgtc cctcatatct cctccttatg gtgttttact aaacttttcc
     3301 atgttctaat cctcatcctg tctacttgcc acacaatcat cacctgccat ctgttttcca
     3361 taatccctaa tgatctttgc ttttcttctt ggcactactt ttatgtcact attatcttgt
     3421 attactactg ccccttcacc tttccagagg agctttgctg gtcctttcca aagtggattt
     3481 ctgctgtccc tgtaataaac ccgaaaattt tgaatttttg taatttgttt ttgtaattct
     3541 ttagtttgta tgtctgttgc tattatgtct actattcttt cccctgcact gtacccccca
     3601 atcccccctt ttcttttaaa attgtggatg aatactgcca tttgtactgc tgtcttaaga
     3661 tgttcagcct gatctcttac ctgtcctata attttcttta attctttatt catagattct
     3721 actactcctt gactttgggg attgtaggga atttccaggt ctgaagatcc tgctgcttgc
     3781 tgtgcttttt tcttactttt gttttgctct tcctctatct tgtctaaagc ttccttggtg
     3841 tcttttatct ctatcctttg atgcacacaa tagagggttg ctactgtatt atataatgat
     3901 ctaagttctt ctgatcctgt ctgaagggat ggttgtagct gtcccagtat ttgtctacag
     3961 ccttctgatg tttctaacag gccagtatta actgcgaatc gttctagctc cctgcttgcc
     4021 catactatat gttttaattt atattttttc tttccccctg gccttaaccg aattttttcc
     4081 catcgcgatc taattctccc ccgcttaata ctgacgctct cgcacccatc tctctccttc
     4141 tagcctccgc tagtcaaaat ttttggcgta ctcaccagtc gccgcccctc gcctcttgcc
     4201 gtgcgcgctt cagcaagccg agtcctgcgt cgagagagct cctctggttt ccctttcgct
     4261 ttcaagtccc tgttcgggcg ccactgctag agattttcca cactgactaa aagggtctga
     4321 gggatctcta gttaccagag tcacacaaca gacgggcaca cactacttga agcactcaag
     4381 gcaagcttta ttgaggctta agcagtgggt tccctagtta gccagagagc tcccaggctc
     4441 agatctggtc taaccagaga gacccagtac aggcaaaaag cagctgctta tatgcaggat
     4501 ctgagggctc gccactcccc agtcccgccc aggccacgcc tcccatggta atagcgatga
     4561 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat
     4621 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca
     4681 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
     4741 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt
     4801 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat
     4861 atgggctatg aactaatgac cccgtaattg attactatta ataactagtc aataatcaat
     4921 gtcgacccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4981 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5041 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5101 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5161 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5221 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5281 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5341 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5401 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5461 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5521 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5581 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5641 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5701 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5761 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5821 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5881 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5941 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6001 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6061 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6121 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6181 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6241 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6301 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6361 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6421 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6481 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6541 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6601 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6661 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6721 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6781 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     6841 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     6901 attcattaat gcagcggatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     6961 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     7021 tgtaaccatt ataagctgca ataaacaagt tagctttttg caaaagccta ggcctccaaa
     7081 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     7141 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     7201 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcggatccgc
     7261 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7321 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7381 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     7441 cttgggctgc aggtcgaggg atctccataa gagaagaggg acagctatga ctgggagtag
     7501 tcaggagagg aggaaaaatc tggctagtaa aacatgtaag gaaaatttta gggatgttaa
     7561 agaaaaaaat aacacaaaac aaaatat
//
