LOCUS       sequence-488252-        7665 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_023, derived from human protein SPOC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7665)
  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 7665)
  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_Vidr_023
FEATURES             Location/Qualifiers
     source          1..7665
                     /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(2696..3406,3407..3409,3410..3412))
                     /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(2738..2759)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3087..3106
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3346..3367
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3409..3418
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3494..3697)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3523..3543)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3698..4001
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4258..4302)
                     /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    4487..4720
                     /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    5080..5197
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5689..5814
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5861..6118
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6120..6499
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6530..6634
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6635..6703,6704..7495)
                     /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(6853..6872)
                     /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 ccggttcatg ctttgcgagg aaatggacat tctgagccag
     2101 cctcgtcagt tggaccaagg gagctgtcac gcgccggtcc tgggggttcg ggggcagcaa
     2161 gtgcgctgct taagcgctgt aatgggcgtg ggatcgagga tttggtctgt ggacacaata
     2221 acgcctgcaa ttgatgtgac atagtagcca gactttccaa gtggcgcaga agggcttgat
     2281 gaggacaact gtcacgatgg aaatgctgtc cgcgaccgaa tccgtgcccc gcgggagcta
     2341 ctgagtaagg atgctgactc tggcgcaaac cgggttcagg aggccattga ccacgtccgg
     2401 gatgctgcca gttttctggc tcgggccaaa gacgcccacg cccacgcggt ggacgctgcc
     2461 acgcagaaat tccgcgtggc gcgatactgc cacgtccttg tgattgctgc cacgcgccac
     2521 ctggaggcgg tgtgcctttc aaaggcacgt tggggcggcg atcatccggc tgataatagc
     2581 gtttctcaac cttagaattg aacgacacca tcttttgaac cttacctaac caaggagatg
     2641 atcccgccgt gtcccgggaa gcggccgcac ttcccccgga gcctccgctt ccgcccttgt
     2701 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt
     2761 gatcgcgctt ctcgttgggg tctttgctca gcttggactg ggtgctcagg tagtggttgt
     2821 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct
     2881 gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct
     2941 gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca
     3001 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt
     3061 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg
     3121 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt
     3181 ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc
     3241 agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg
     3301 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct
     3361 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga
     3421 ccggtggatc ccgggcccgc ggtaccgtcg actgcagaat tcggttcact aaacgagccc
     3481 ggttcactaa acgagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
     3541 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
     3601 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     3661 atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata
     3721 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     3781 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     3841 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     3901 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     3961 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     4021 tatgaactaa tgaccccgta attgattact attaataact agcacgcgtt cgaagcttga
     4081 gctcggtccc ctcggggttg ggaggtgggt ctgaaacgat aatggtgaat atccctgcct
     4141 aactctattc actatagaaa gtacagcaaa aactattctt aaacctacca agcctcctac
     4201 tatcattatg aataatttta tataccacag ccaatttgtt atgttaaacc aattccacaa
     4261 acttgcccat ttatctaatt ccaataattc ttgttcattc ttttcttgct ggttttgcga
     4321 ttcttcaatt aaggagtgta ttaagcttgt gtaattgtta atttctctgt cccactccat
     4381 ccaggtcgtg tgattccaaa tctgttccag agatttatta ctccaactag cattccaagg
     4441 cacagcagtg gtgcaaatga gttttccaga gcaaccccaa atccccagga gctgttgatc
     4501 ctttaggtat ctttccacag ccaggattct tgcctggagc tgcttgatgc cccagactgt
     4561 gagttgcaac agatgctgtt gcgcctcaat agccctcagc aaattgttct gctgctgcac
     4621 tataccagac aataattgtc tggcctgtac cgtcagcgtc attgacgctg cgcccatagt
     4681 gcttcctgct gctcccaaga acccaaggaa caaagctcct attcccactg ctcttttttc
     4741 tctctgcacc actcttctct ttgccttggt gggtgctact cctaatggtt caatttttac
     4801 tactttatat ttatataatt cacttctcca attgtccctc atatctcctc cttatggtgt
     4861 tttactaaac ttttccatgt tctaatcctc atcctgtcta cttgccacac aatcatcacc
     4921 tgccatctgt tttccataat ccctaatgat ctttgctttt cttcttggca ctacttttat
     4981 gtcactatta tcttgtatta ctactgcccc ttcacctttc cagaggagct ttgctggtcc
     5041 tttccaaagt ggatttctgc tgtccctgta ataaacccga aaattttgaa tttttgtaat
     5101 ttgtttttgt aattctttag tttgtatgtc tgttgctatt atgtctacta ttctttcccc
     5161 tgcactgtac cccccaatcc ccccttttct tttaaaattg tggatgaata ctgccatttg
     5221 tactgctgtc ttaagatgtt cagcctgatc tcttacctgt cctataattt tctttaattc
     5281 tttattcata gattctacta ctccttgact ttggggattg tagggaattt ccaggtctga
     5341 agatcctgct gcttgctgtg cttttttctt acttttgttt tgctcttcct ctatcttgtc
     5401 taaagcttcc ttggtgtctt ttatctctat cctttgatgc acacaataga gggttgctac
     5461 tgtattatat aatgatctaa gttcttctga tcctgtctga agggatggtt gtagctgtcc
     5521 cagtatttgt ctacagcctt ctgatgtttc taacaggcca gtattaactg cgaatcgttc
     5581 tagctccctg cttgcccata ctatatgttt taatttatat tttttctttc cccctggcct
     5641 taaccgaatt ttttcccatc gcgatctaat tctcccccgc ttaatactga cgctctcgca
     5701 cccatctctc tccttctagc ctccgctagt caaaattttt ggcgtactca ccagtcgccg
     5761 cccctcgcct cttgccgtgc gcgcttcagc aagccgagtc ctgcgtcgag agagctcctc
     5821 tggtttccct ttcgctttca agtccctgtt cgggcgccac tgctagagat tttccacact
     5881 gactaaaagg gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact
     5941 acttgaagca ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca
     6001 gagagctccc aggctcagat ctggtctaac cagagagacc cagtacaggc aaaaagcagc
     6061 tgcttatatg caggatctga gggctcgcca ctccccagtc ccgcccaggc cacgcctccc
     6121 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
     6181 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
     6241 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     6301 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     6361 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
     6421 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa
     6481 ctagtcaata atcaatgtcg acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     6541 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     6601 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     6661 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6721 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     6781 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     6841 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     6901 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     6961 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     7021 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     7081 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     7141 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     7201 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     7261 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     7321 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     7381 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     7441 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     7501 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     7561 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7621 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
