LOCUS       sequence-488219-        7455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_202, derived from human fusion protein TTC6_MIPOL1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7455)
  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 7455)
  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_202
FEATURES             Location/Qualifiers
     source          1..7455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    828..962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(852..871)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     906..925
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(968..1163)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      982..1117
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    1222..1243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1258..1263,1264..1281,1282..1288)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1296..1312
                     /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     1301..1323
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1320..1336
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1320..1336
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1679..1698
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1697..1752
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1798..1817
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(2486..3196,3197..3199,3200..3202))
                     /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(2528..2549)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2877..2896
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3136..3157
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3199..3208
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3284..3487)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3313..3333)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3488..3791
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4048..4092)
                     /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    4277..4510
                     /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    4870..4987
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5479..5604
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5651..5908
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5910..6289
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6320..6424
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6425..6493,6494..7285)
                     /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(6643..6662)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagcggat ccagacatga
      841 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      901 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      961 ttagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     1021 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     1081 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     1141 tggttgctga ctaattgaga tgcggatccg ctggcacgac aggtttcccg actggaaagc
     1201 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     1261 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     1321 aggaaacagc tatgaccatg attacgccaa gcttgggctg caggtcgagg gatctccata
     1381 agagaagagg gacagctatg actgggagta gtcaggagag gaggaaaaat ctggctagta
     1441 aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa taacacaaaa caaaatataa
     1501 aaaaaatcta acctcaagtc aaggcttttc tatggaataa ggaatggaca gcagggggct
     1561 gtttcatata ctgatgacct ctttatagcc aacctttgtt catggcagcc agcatatggg
     1621 catatgttgc caaactctaa accaaatact cattctgatg ttttaaatga tttgccctcc
     1681 catatgtcct tccgagtgag agacacaaaa aattccaaca cactattgca atgaaaataa
     1741 atttccttta ttagccagaa gtcagatgct caaggggctt catgatgtcc ccataatttt
     1801 tggcagaggg aaaaagatct gctagctcga gcagctgaag ctttattgag gcttaagcag
     1861 tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc
     1921 agtacaggca aaaagcagat cttgtcttct ttgggagtga attagccctt ccagtccccc
     1981 cttttctttt aaaaagtggc taagatctac agctgccttg taagtcattg gtcttaaagg
     2041 taccgtaatc ttatctagaa ccggttcacc aagcagcttc gcgcgcatca gatggtggcc
     2101 atgattcctg ctccccctgt gcgcctaaac cagcagcctc gcgtgcctca cgggcccccg
     2161 actcagacac aaaattactg cgaatacgga ctttacgacg cccgctggaa gcttccatat
     2221 aaccgtcttc ggcggaaaca cttgatgcct tacgttcttt ggccatacac gggccagcca
     2281 gaggagctac ctcttgagac tgttcacgtt cacgacctaa atggaatacg cggcgtggtg
     2341 cagcgtggcg agatgactcc agcacaccac gcgcgcgacg cttaaccggt ggctctggtg
     2401 gcggtaaaag gacgaccggg cgggcagtac caaagccact tgcccgggaa gcggccgcac
     2461 ttcccccgga gcctccgctt ccgcccttgt acagctcgtc catgccgaga gtgatcccgg
     2521 cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca
     2581 gcttggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg
     2641 gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga
     2701 tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag acgttgtggc
     2761 tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc
     2821 ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct
     2881 tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg
     2941 cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag cactgcacgc
     3001 cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga
     3061 tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga
     3121 acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca
     3181 gctcctcgcc cttgctcacc atggtggcga ccggtggatc ccgggcccgc ggtaccgtcg
     3241 actgcagaat tcggttcact aaacgagccc ggttcactaa acgagctctg cttatataga
     3301 cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt
     3361 ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga
     3421 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc
     3481 gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     3541 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     3601 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     3661 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     3721 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     3781 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
     3841 attaataact agcacgcgtt cgaagcttga gctcggtccc ctcggggttg ggaggtgggt
     3901 ctgaaacgat aatggtgaat atccctgcct aactctattc actatagaaa gtacagcaaa
     3961 aactattctt aaacctacca agcctcctac tatcattatg aataatttta tataccacag
     4021 ccaatttgtt atgttaaacc aattccacaa acttgcccat ttatctaatt ccaataattc
     4081 ttgttcattc ttttcttgct ggttttgcga ttcttcaatt aaggagtgta ttaagcttgt
     4141 gtaattgtta atttctctgt cccactccat ccaggtcgtg tgattccaaa tctgttccag
     4201 agatttatta ctccaactag cattccaagg cacagcagtg gtgcaaatga gttttccaga
     4261 gcaaccccaa atccccagga gctgttgatc ctttaggtat ctttccacag ccaggattct
     4321 tgcctggagc tgcttgatgc cccagactgt gagttgcaac agatgctgtt gcgcctcaat
     4381 agccctcagc aaattgttct gctgctgcac tataccagac aataattgtc tggcctgtac
     4441 cgtcagcgtc attgacgctg cgcccatagt gcttcctgct gctcccaaga acccaaggaa
     4501 caaagctcct attcccactg ctcttttttc tctctgcacc actcttctct ttgccttggt
     4561 gggtgctact cctaatggtt caatttttac tactttatat ttatataatt cacttctcca
     4621 attgtccctc atatctcctc cttatggtgt tttactaaac ttttccatgt tctaatcctc
     4681 atcctgtcta cttgccacac aatcatcacc tgccatctgt tttccataat ccctaatgat
     4741 ctttgctttt cttcttggca ctacttttat gtcactatta tcttgtatta ctactgcccc
     4801 ttcacctttc cagaggagct ttgctggtcc tttccaaagt ggatttctgc tgtccctgta
     4861 ataaacccga aaattttgaa tttttgtaat ttgtttttgt aattctttag tttgtatgtc
     4921 tgttgctatt atgtctacta ttctttcccc tgcactgtac cccccaatcc ccccttttct
     4981 tttaaaattg tggatgaata ctgccatttg tactgctgtc ttaagatgtt cagcctgatc
     5041 tcttacctgt cctataattt tctttaattc tttattcata gattctacta ctccttgact
     5101 ttggggattg tagggaattt ccaggtctga agatcctgct gcttgctgtg cttttttctt
     5161 acttttgttt tgctcttcct ctatcttgtc taaagcttcc ttggtgtctt ttatctctat
     5221 cctttgatgc acacaataga gggttgctac tgtattatat aatgatctaa gttcttctga
     5281 tcctgtctga agggatggtt gtagctgtcc cagtatttgt ctacagcctt ctgatgtttc
     5341 taacaggcca gtattaactg cgaatcgttc tagctccctg cttgcccata ctatatgttt
     5401 taatttatat tttttctttc cccctggcct taaccgaatt ttttcccatc gcgatctaat
     5461 tctcccccgc ttaatactga cgctctcgca cccatctctc tccttctagc ctccgctagt
     5521 caaaattttt ggcgtactca ccagtcgccg cccctcgcct cttgccgtgc gcgcttcagc
     5581 aagccgagtc ctgcgtcgag agagctcctc tggtttccct ttcgctttca agtccctgtt
     5641 cgggcgccac tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta
     5701 ccagagtcac acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga
     5761 ggcttaagca gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac
     5821 cagagagacc cagtacaggc aaaaagcagc tgcttatatg caggatctga gggctcgcca
     5881 ctccccagtc ccgcccaggc cacgcctccc atggtaatag cgatgactaa tacgtagatg
     5941 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
     6001 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
     6061 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
     6121 gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag
     6181 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
     6241 aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtcg acccaggtgg
     6301 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     6361 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     6421 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     6481 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     6541 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     6601 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     6661 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     6721 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     6781 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6841 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6901 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6961 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     7021 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     7081 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     7141 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     7201 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     7261 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     7321 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     7381 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     7441 gatcaaagga tcttc
//
