LOCUS       sequence-488248-        7404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_019, derived from human protein TRIPB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7404)
  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 7404)
  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_019
FEATURES             Location/Qualifiers
     source          1..7404
                     /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(2435..3145,3146..3148,3149..3151))
                     /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(2477..2498)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2826..2845
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3085..3106
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3148..3157
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3233..3436)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3262..3282)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3437..3740
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3997..4041)
                     /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    4226..4459
                     /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    4819..4936
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5428..5553
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5600..5857
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5859..6238
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6269..6373
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6374..6442,6443..7234)
                     /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(6592..6611)
                     /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 ccggttcatt gcttcagtaa gtctttcaga actacgcctg
     2101 ccgaattatc aggaagggca ggcaacggtg tgaacgtcgg taatacgtca ctaattggct
     2161 ttaacaacag atgacccgga ccccctggtc ctaacccggc cggattgatt aagggtactg
     2221 ctgcgctgcg gggggcaaga aagggattta cgtcagtacg acgtccgctg cgtgattcag
     2281 ctgtatcctt aaaggattca ggtgcgttcg tgtcgcgctt gcggcggccg ggactgtcca
     2341 agggtttcat atcgtgcacg ctcaacttcg gcgggggaat actgggatgg ctccgggaag
     2401 cggccgcact tcccccggag cctccgcttc cgcccttgta cagctcgtcc atgccgagag
     2461 tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt
     2521 ctttgctcag cttggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt
     2581 cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt
     2641 tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga
     2701 cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga
     2761 agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg
     2821 cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt
     2881 cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc
     2941 actgcacgcc gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg
     3001 tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg
     3061 acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc
     3121 cggtgaacag ctcctcgccc ttgctcacca tggtggcgac cggtggatcc cgggcccgcg
     3181 gtaccgtcga ctgcagaatt cggttcacta aacgagcccg gttcactaaa cgagctctgc
     3241 ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt
     3301 tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa
     3361 tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact
     3421 gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     3481 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     3541 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     3601 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     3661 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     3721 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     3781 ttgattacta ttaataacta gcacgcgttc gaagcttgag ctcggtcccc tcggggttgg
     3841 gaggtgggtc tgaaacgata atggtgaata tccctgccta actctattca ctatagaaag
     3901 tacagcaaaa actattctta aacctaccaa gcctcctact atcattatga ataattttat
     3961 ataccacagc caatttgtta tgttaaacca attccacaaa cttgcccatt tatctaattc
     4021 caataattct tgttcattct tttcttgctg gttttgcgat tcttcaatta aggagtgtat
     4081 taagcttgtg taattgttaa tttctctgtc ccactccatc caggtcgtgt gattccaaat
     4141 ctgttccaga gatttattac tccaactagc attccaaggc acagcagtgg tgcaaatgag
     4201 ttttccagag caaccccaaa tccccaggag ctgttgatcc tttaggtatc tttccacagc
     4261 caggattctt gcctggagct gcttgatgcc ccagactgtg agttgcaaca gatgctgttg
     4321 cgcctcaata gccctcagca aattgttctg ctgctgcact ataccagaca ataattgtct
     4381 ggcctgtacc gtcagcgtca ttgacgctgc gcccatagtg cttcctgctg ctcccaagaa
     4441 cccaaggaac aaagctccta ttcccactgc tcttttttct ctctgcacca ctcttctctt
     4501 tgccttggtg ggtgctactc ctaatggttc aatttttact actttatatt tatataattc
     4561 acttctccaa ttgtccctca tatctcctcc ttatggtgtt ttactaaact tttccatgtt
     4621 ctaatcctca tcctgtctac ttgccacaca atcatcacct gccatctgtt ttccataatc
     4681 cctaatgatc tttgcttttc ttcttggcac tacttttatg tcactattat cttgtattac
     4741 tactgcccct tcacctttcc agaggagctt tgctggtcct ttccaaagtg gatttctgct
     4801 gtccctgtaa taaacccgaa aattttgaat ttttgtaatt tgtttttgta attctttagt
     4861 ttgtatgtct gttgctatta tgtctactat tctttcccct gcactgtacc ccccaatccc
     4921 cccttttctt ttaaaattgt ggatgaatac tgccatttgt actgctgtct taagatgttc
     4981 agcctgatct cttacctgtc ctataatttt ctttaattct ttattcatag attctactac
     5041 tccttgactt tggggattgt agggaatttc caggtctgaa gatcctgctg cttgctgtgc
     5101 ttttttctta cttttgtttt gctcttcctc tatcttgtct aaagcttcct tggtgtcttt
     5161 tatctctatc ctttgatgca cacaatagag ggttgctact gtattatata atgatctaag
     5221 ttcttctgat cctgtctgaa gggatggttg tagctgtccc agtatttgtc tacagccttc
     5281 tgatgtttct aacaggccag tattaactgc gaatcgttct agctccctgc ttgcccatac
     5341 tatatgtttt aatttatatt ttttctttcc ccctggcctt aaccgaattt tttcccatcg
     5401 cgatctaatt ctcccccgct taatactgac gctctcgcac ccatctctct ccttctagcc
     5461 tccgctagtc aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg
     5521 cgcttcagca agccgagtcc tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa
     5581 gtccctgttc gggcgccact gctagagatt ttccacactg actaaaaggg tctgagggat
     5641 ctctagttac cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag
     5701 ctttattgag gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc
     5761 tggtctaacc agagagaccc agtacaggca aaaagcagct gcttatatgc aggatctgag
     5821 ggctcgccac tccccagtcc cgcccaggcc acgcctccca tggtaatagc gatgactaat
     5881 acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag
     5941 gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga
     6001 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
     6061 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
     6121 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
     6181 ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcga
     6241 cccaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     6301 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     6361 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     6421 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     6481 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     6541 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     6601 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     6661 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     6721 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     6781 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     6841 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     6901 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     6961 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     7021 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     7081 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     7141 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     7201 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     7261 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     7321 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     7381 cgtagaaaag atcaaaggat cttc
//
