LOCUS       sequence-488249-        7560 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_020, derived from human protein NPM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7560)
  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 7560)
  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_020
FEATURES             Location/Qualifiers
     source          1..7560
                     /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(2591..3301,3302..3304,3305..3307))
                     /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(2633..2654)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2982..3001
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3241..3262
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3304..3313
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3389..3592)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3418..3438)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3593..3896
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4153..4197)
                     /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    4382..4615
                     /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    4975..5092
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5584..5709
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5756..6013
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6015..6394
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6425..6529
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6530..6598,6599..7390)
                     /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(6748..6767)
                     /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 tggcttcaac ctttggtaag ctcccccctt
     2101 tctcgattga tgcctgcatt ttggccttga tatcttctac agaactaggc cccttaggcg
     2161 tcttaggagt tttttcttgt tttttgaagc tctcttgccc cttcgaacgc ggagtgctag
     2221 atggcttcga atccttcccg ttttggttac tcttttgagc attcttagca ggagtatcgc
     2281 ggatggattt tttgaccggc gctttttcct cagcttcctc atcatcaaaa tcatcgtcgt
     2341 catcatcttc atcatcgtct tcttcgtcgt catcatcgtc gtcctcgtcc gctgctaact
     2401 taactttctt ttggggtact ttggacccac ctccgggtgc gctgcgcttc cctgagatgc
     2461 tcaataattt cacatcctct tcttcttcat cctcggactc ggcgtcttcc tccactgcca
     2521 ccaagtgctg gccagagatg tgaactggcc gggaagcggc cgcacttccc ccggagcctc
     2581 cgcttccgcc cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact
     2641 ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagcttg gactgggtgc
     2701 tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt
     2761 agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg aagttcacct
     2821 tgatgccgtt cttctgcttg tcggccatga tatagacgtt gtggctgttg tagttgtact
     2881 ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc
     2941 ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt
     3001 ccttgaagaa gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt
     3061 cgtgctgctt catgtggtcg gggtagcggc tgaagcactg cacgccgtag gtcagggtgg
     3121 tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca
     3181 gcttgccgta ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta
     3241 cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc
     3301 tcaccatggt ggcgaccggt ggatcccggg cccgcggtac cgtcgactgc agaattcggt
     3361 tcactaaacg agcccggttc actaaacgag ctctgcttat atagacctcc caccgtacac
     3421 gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt
     3481 gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc
     3541 gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa
     3601 tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact
     3661 gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca
     3721 tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac
     3781 gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg
     3841 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga
     3901 actccatata tgggctatga actaatgacc ccgtaattga ttactattaa taactagcac
     3961 gcgttcgaag cttgagctcg gtcccctcgg ggttgggagg tgggtctgaa acgataatgg
     4021 tgaatatccc tgcctaactc tattcactat agaaagtaca gcaaaaacta ttcttaaacc
     4081 taccaagcct cctactatca ttatgaataa ttttatatac cacagccaat ttgttatgtt
     4141 aaaccaattc cacaaacttg cccatttatc taattccaat aattcttgtt cattcttttc
     4201 ttgctggttt tgcgattctt caattaagga gtgtattaag cttgtgtaat tgttaatttc
     4261 tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt tccagagatt tattactcca
     4321 actagcattc caaggcacag cagtggtgca aatgagtttt ccagagcaac cccaaatccc
     4381 caggagctgt tgatccttta ggtatctttc cacagccagg attcttgcct ggagctgctt
     4441 gatgccccag actgtgagtt gcaacagatg ctgttgcgcc tcaatagccc tcagcaaatt
     4501 gttctgctgc tgcactatac cagacaataa ttgtctggcc tgtaccgtca gcgtcattga
     4561 cgctgcgccc atagtgcttc ctgctgctcc caagaaccca aggaacaaag ctcctattcc
     4621 cactgctctt ttttctctct gcaccactct tctctttgcc ttggtgggtg ctactcctaa
     4681 tggttcaatt tttactactt tatatttata taattcactt ctccaattgt ccctcatatc
     4741 tcctccttat ggtgttttac taaacttttc catgttctaa tcctcatcct gtctacttgc
     4801 cacacaatca tcacctgcca tctgttttcc ataatcccta atgatctttg cttttcttct
     4861 tggcactact tttatgtcac tattatcttg tattactact gccccttcac ctttccagag
     4921 gagctttgct ggtcctttcc aaagtggatt tctgctgtcc ctgtaataaa cccgaaaatt
     4981 ttgaattttt gtaatttgtt tttgtaattc tttagtttgt atgtctgttg ctattatgtc
     5041 tactattctt tcccctgcac tgtacccccc aatcccccct tttcttttaa aattgtggat
     5101 gaatactgcc atttgtactg ctgtcttaag atgttcagcc tgatctctta cctgtcctat
     5161 aattttcttt aattctttat tcatagattc tactactcct tgactttggg gattgtaggg
     5221 aatttccagg tctgaagatc ctgctgcttg ctgtgctttt ttcttacttt tgttttgctc
     5281 ttcctctatc ttgtctaaag cttccttggt gtcttttatc tctatccttt gatgcacaca
     5341 atagagggtt gctactgtat tatataatga tctaagttct tctgatcctg tctgaaggga
     5401 tggttgtagc tgtcccagta tttgtctaca gccttctgat gtttctaaca ggccagtatt
     5461 aactgcgaat cgttctagct ccctgcttgc ccatactata tgttttaatt tatatttttt
     5521 ctttccccct ggccttaacc gaattttttc ccatcgcgat ctaattctcc cccgcttaat
     5581 actgacgctc tcgcacccat ctctctcctt ctagcctccg ctagtcaaaa tttttggcgt
     5641 actcaccagt cgccgcccct cgcctcttgc cgtgcgcgct tcagcaagcc gagtcctgcg
     5701 tcgagagagc tcctctggtt tccctttcgc tttcaagtcc ctgttcgggc gccactgcta
     5761 gagattttcc acactgacta aaagggtctg agggatctct agttaccaga gtcacacaac
     5821 agacgggcac acactacttg aagcactcaa ggcaagcttt attgaggctt aagcagtggg
     5881 ttccctagtt agccagagag ctcccaggct cagatctggt ctaaccagag agacccagta
     5941 caggcaaaaa gcagctgctt atatgcagga tctgagggct cgccactccc cagtcccgcc
     6001 caggccacgc ctcccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga
     6061 aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg
     6121 tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac
     6181 cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca
     6241 tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt
     6301 accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt
     6361 gattactatt aataactagt caataatcaa tgtcgaccca ggtggcactt ttcggggaaa
     6421 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     6481 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     6541 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     6601 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     6661 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     6721 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     6781 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     6841 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6901 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     6961 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     7021 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     7081 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     7141 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     7201 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     7261 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     7321 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     7381 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     7441 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     7501 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
