LOCUS       sequence-488221-        7323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_204, derived from human fusion protein TTC6_MIPOL1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7323)
  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 7323)
  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_204
FEATURES             Location/Qualifiers
     source          1..7323
                     /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(856..1566,1567..1569,1570..1572))
                     /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(898..919)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1247..1266
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1506..1527
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1569..1578
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1654..1857)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1683..1703)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1858..2161
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2418..2462)
                     /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    2647..2880
                     /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    3240..3357
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3849..3974
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4021..4278
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4280..4659
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4690..4794
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4795..4863,4864..5655)
                     /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(5013..5032)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5826..6414
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6315..6334
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6568..6585
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6653..6787
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6677..6696)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6731..6750
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6793..6988)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6807..6942
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7047..7068
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7083..7088,7089..7106,7107..7113)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7121..7137
                     /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     7126..7148
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7145..7161
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7145..7161
                     /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 aaaagagcgc gtcttaccag gtgcagcggc
      601 ttcccctaac ccgctcggac gcggagcacc tccaagcact tcttgcagaa gtgcagcggc
      661 gctcattgcc ggacctttta acgatgggta acggcgcgat gtacgggctg ccgcgcatga
      721 tacgcgggcc gggcgtgcag gacctggggc atgtaaggaa tgcactggcg actcatcaac
      781 tgctggcttc gaggccaact tctgcttaaa gcgccgggaa gcggccgcac ttcccccgga
      841 gcctccgctt ccgcccttgt acagctcgtc catgccgaga gtgatcccgg cggcggtcac
      901 gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca gcttggactg
      961 ggtgctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg gggtgttctg
     1021 ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt
     1081 caccttgatg ccgttcttct gcttgtcggc catgatatag acgttgtggc tgttgtagtt
     1141 gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc
     1201 gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc
     1261 gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa
     1321 gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag cactgcacgc cgtaggtcag
     1381 ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga tgaacttcag
     1441 ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc
     1501 gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc
     1561 cttgctcacc atggtggcga ccggtggatc ccgggcccgc ggtaccgtcg actgcagaat
     1621 tcggttcact aaacgagccc ggttcactaa acgagctctg cttatataga cctcccaccg
     1681 tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc
     1741 ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa
     1801 tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc gcatcaccat
     1861 ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat
     1921 gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact
     1981 tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca
     2041 ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt
     2101 caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac
     2161 gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact attaataact
     2221 agcacgcgtt cgaagcttga gctcggtccc ctcggggttg ggaggtgggt ctgaaacgat
     2281 aatggtgaat atccctgcct aactctattc actatagaaa gtacagcaaa aactattctt
     2341 aaacctacca agcctcctac tatcattatg aataatttta tataccacag ccaatttgtt
     2401 atgttaaacc aattccacaa acttgcccat ttatctaatt ccaataattc ttgttcattc
     2461 ttttcttgct ggttttgcga ttcttcaatt aaggagtgta ttaagcttgt gtaattgtta
     2521 atttctctgt cccactccat ccaggtcgtg tgattccaaa tctgttccag agatttatta
     2581 ctccaactag cattccaagg cacagcagtg gtgcaaatga gttttccaga gcaaccccaa
     2641 atccccagga gctgttgatc ctttaggtat ctttccacag ccaggattct tgcctggagc
     2701 tgcttgatgc cccagactgt gagttgcaac agatgctgtt gcgcctcaat agccctcagc
     2761 aaattgttct gctgctgcac tataccagac aataattgtc tggcctgtac cgtcagcgtc
     2821 attgacgctg cgcccatagt gcttcctgct gctcccaaga acccaaggaa caaagctcct
     2881 attcccactg ctcttttttc tctctgcacc actcttctct ttgccttggt gggtgctact
     2941 cctaatggtt caatttttac tactttatat ttatataatt cacttctcca attgtccctc
     3001 atatctcctc cttatggtgt tttactaaac ttttccatgt tctaatcctc atcctgtcta
     3061 cttgccacac aatcatcacc tgccatctgt tttccataat ccctaatgat ctttgctttt
     3121 cttcttggca ctacttttat gtcactatta tcttgtatta ctactgcccc ttcacctttc
     3181 cagaggagct ttgctggtcc tttccaaagt ggatttctgc tgtccctgta ataaacccga
     3241 aaattttgaa tttttgtaat ttgtttttgt aattctttag tttgtatgtc tgttgctatt
     3301 atgtctacta ttctttcccc tgcactgtac cccccaatcc ccccttttct tttaaaattg
     3361 tggatgaata ctgccatttg tactgctgtc ttaagatgtt cagcctgatc tcttacctgt
     3421 cctataattt tctttaattc tttattcata gattctacta ctccttgact ttggggattg
     3481 tagggaattt ccaggtctga agatcctgct gcttgctgtg cttttttctt acttttgttt
     3541 tgctcttcct ctatcttgtc taaagcttcc ttggtgtctt ttatctctat cctttgatgc
     3601 acacaataga gggttgctac tgtattatat aatgatctaa gttcttctga tcctgtctga
     3661 agggatggtt gtagctgtcc cagtatttgt ctacagcctt ctgatgtttc taacaggcca
     3721 gtattaactg cgaatcgttc tagctccctg cttgcccata ctatatgttt taatttatat
     3781 tttttctttc cccctggcct taaccgaatt ttttcccatc gcgatctaat tctcccccgc
     3841 ttaatactga cgctctcgca cccatctctc tccttctagc ctccgctagt caaaattttt
     3901 ggcgtactca ccagtcgccg cccctcgcct cttgccgtgc gcgcttcagc aagccgagtc
     3961 ctgcgtcgag agagctcctc tggtttccct ttcgctttca agtccctgtt cgggcgccac
     4021 tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac
     4081 acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca
     4141 gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc
     4201 cagtacaggc aaaaagcagc tgcttatatg caggatctga gggctcgcca ctccccagtc
     4261 ccgcccaggc cacgcctccc atggtaatag cgatgactaa tacgtagatg tactgccaag
     4321 taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
     4381 tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag
     4441 tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg
     4501 gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc
     4561 catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg
     4621 taattgatta ctattaataa ctagtcaata atcaatgtcg acccaggtgg cacttttcgg
     4681 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4741 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4801 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4861 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4921 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4981 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5041 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5101 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5161 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5221 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5281 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5341 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5401 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5461 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5521 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5581 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5641 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5701 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5761 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5821 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5881 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5941 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6001 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6061 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6121 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6181 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6241 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6301 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6361 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6421 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6481 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     6541 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     6601 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag cggatccaga
     6661 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     6721 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     6781 acaagttagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     6841 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     6901 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     6961 gactatggtt gctgactaat tgagatgcgg atccgctggc acgacaggtt tcccgactgg
     7021 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     7081 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     7141 cacacaggaa acagctatga ccatgattac gccaagcttg ggctgcaggt cgagggatct
     7201 ccataagaga agagggacag ctatgactgg gagtagtcag gagaggagga aaaatctggc
     7261 tagtaaaaca tgtaaggaaa attttaggga tgttaaagaa aaaaataaca caaaacaaaa
     7321 tat
//
