LOCUS       sequence-494365-        7737 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_142, derived from human fusion protein PAX5_NCOA5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7737)
  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 7737)
  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_142
FEATURES             Location/Qualifiers
     source          1..7737
                     /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(1270..1980,1981..1983,1984..1986))
                     /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(1312..1333)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1661..1680
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1920..1941
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1983..1992
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2068..2271)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2097..2117)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2272..2575
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2832..2876)
                     /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    3061..3294
                     /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    3654..3771
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4263..4388
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4435..4692
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4694..5073
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5104..5208
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5209..5277,5278..6069)
                     /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(5427..5446)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6240..6828
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6729..6748
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6982..6999
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7067..7201
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7091..7110)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7145..7164
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7207..7402)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7221..7356
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7461..7482
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7497..7502,7503..7520,7521..7527)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7535..7551
                     /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     7540..7562
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7559..7575
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7559..7575
                     /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 gtagtgacgt tgatagctgc ccatcggggc
      601 ctgtggttga cccatttgcg cggttgtctg gctgacaaga tgagataagg caggtccaga
      661 ctgaatcaag gtgtcaagcg ctttttggac actaggatta tcgaaattga tcccagtgga
      721 ggagactggg cgttgcgaag tcatgttgct agcgggagca aggcgacttg aaggctgacc
      781 aaacagtcct tgtgatggag ctcccggacg aggacccatg ttttgagcgc tccctccttg
      841 gcccaaaata ctaggcggct ggtttccact ggcttgcgaa cgctgctgtg gttggctgtt
      901 cgcagcagtt gagaagtttt gattcggggt gttcccggct gcaacagatg gcgaggcact
      961 ggaggagttt gcggtcaccg tgcctgaatt gaataaagac agaatcttgg cctgaagctc
     1021 ttgttgcgat gtcggtggcg cactcggtgt cggcgtcgcg gaaggtaaaa cttgaccact
     1081 ttgcaagggc tggctacttg gttgggtttt caaggatgca cctgacgtgg cccccaatgg
     1141 ttggcgagaa atcggccctg gtaaagagtc tgtggaggag cgcatcaggc gctccttacg
     1201 ttcacgcaag tagttaataa ttttatcccg ggaagcggcc gcacttcccc cggagcctcc
     1261 gcttccgccc ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc
     1321 cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg actgggtgct
     1381 caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta
     1441 gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt
     1501 gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc
     1561 cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg
     1621 gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc
     1681 cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc
     1741 gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt
     1801 cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag
     1861 cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac
     1921 gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct
     1981 caccatggtg gcgaccggtg gatcccgggc ccgcggtacc gtcgactgca gaattcggtt
     2041 cactaaacga gcccggttca ctaaacgagc tctgcttata tagacctccc accgtacacg
     2101 cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg
     2161 attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg
     2221 tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat
     2281 agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg
     2341 ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat
     2401 atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg
     2461 tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg
     2521 gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa
     2581 ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagcacg
     2641 cgttcgaagc ttgagctcgg tcccctcggg gttgggaggt gggtctgaaa cgataatggt
     2701 gaatatccct gcctaactct attcactata gaaagtacag caaaaactat tcttaaacct
     2761 accaagcctc ctactatcat tatgaataat tttatatacc acagccaatt tgttatgtta
     2821 aaccaattcc acaaacttgc ccatttatct aattccaata attcttgttc attcttttct
     2881 tgctggtttt gcgattcttc aattaaggag tgtattaagc ttgtgtaatt gttaatttct
     2941 ctgtcccact ccatccaggt cgtgtgattc caaatctgtt ccagagattt attactccaa
     3001 ctagcattcc aaggcacagc agtggtgcaa atgagttttc cagagcaacc ccaaatcccc
     3061 aggagctgtt gatcctttag gtatctttcc acagccagga ttcttgcctg gagctgcttg
     3121 atgccccaga ctgtgagttg caacagatgc tgttgcgcct caatagccct cagcaaattg
     3181 ttctgctgct gcactatacc agacaataat tgtctggcct gtaccgtcag cgtcattgac
     3241 gctgcgccca tagtgcttcc tgctgctccc aagaacccaa ggaacaaagc tcctattccc
     3301 actgctcttt tttctctctg caccactctt ctctttgcct tggtgggtgc tactcctaat
     3361 ggttcaattt ttactacttt atatttatat aattcacttc tccaattgtc cctcatatct
     3421 cctccttatg gtgttttact aaacttttcc atgttctaat cctcatcctg tctacttgcc
     3481 acacaatcat cacctgccat ctgttttcca taatccctaa tgatctttgc ttttcttctt
     3541 ggcactactt ttatgtcact attatcttgt attactactg ccccttcacc tttccagagg
     3601 agctttgctg gtcctttcca aagtggattt ctgctgtccc tgtaataaac ccgaaaattt
     3661 tgaatttttg taatttgttt ttgtaattct ttagtttgta tgtctgttgc tattatgtct
     3721 actattcttt cccctgcact gtacccccca atcccccctt ttcttttaaa attgtggatg
     3781 aatactgcca tttgtactgc tgtcttaaga tgttcagcct gatctcttac ctgtcctata
     3841 attttcttta attctttatt catagattct actactcctt gactttgggg attgtaggga
     3901 atttccaggt ctgaagatcc tgctgcttgc tgtgcttttt tcttactttt gttttgctct
     3961 tcctctatct tgtctaaagc ttccttggtg tcttttatct ctatcctttg atgcacacaa
     4021 tagagggttg ctactgtatt atataatgat ctaagttctt ctgatcctgt ctgaagggat
     4081 ggttgtagct gtcccagtat ttgtctacag ccttctgatg tttctaacag gccagtatta
     4141 actgcgaatc gttctagctc cctgcttgcc catactatat gttttaattt atattttttc
     4201 tttccccctg gccttaaccg aattttttcc catcgcgatc taattctccc ccgcttaata
     4261 ctgacgctct cgcacccatc tctctccttc tagcctccgc tagtcaaaat ttttggcgta
     4321 ctcaccagtc gccgcccctc gcctcttgcc gtgcgcgctt cagcaagccg agtcctgcgt
     4381 cgagagagct cctctggttt ccctttcgct ttcaagtccc tgttcgggcg ccactgctag
     4441 agattttcca cactgactaa aagggtctga gggatctcta gttaccagag tcacacaaca
     4501 gacgggcaca cactacttga agcactcaag gcaagcttta ttgaggctta agcagtgggt
     4561 tccctagtta gccagagagc tcccaggctc agatctggtc taaccagaga gacccagtac
     4621 aggcaaaaag cagctgctta tatgcaggat ctgagggctc gccactcccc agtcccgccc
     4681 aggccacgcc tcccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     4741 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     4801 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     4861 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     4921 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     4981 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     5041 attactatta ataactagtc aataatcaat gtcgacccag gtggcacttt tcggggaaat
     5101 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5161 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5221 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5281 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5341 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5401 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5461 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5521 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5581 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5641 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5701 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5761 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5821 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5881 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5941 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6001 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6061 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6121 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6181 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6241 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6301 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6361 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6421 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6481 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6541 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6601 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6661 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6721 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6781 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6841 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6901 ttatcccctg cttctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     6961 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     7021 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagcggatc cagacatgat
     7081 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     7141 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     7201 tagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     7261 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     7321 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     7381 ggttgctgac taattgagat gcggatccgc tggcacgaca ggtttcccga ctggaaagcg
     7441 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     7501 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     7561 ggaaacagct atgaccatga ttacgccaag cttgggctgc aggtcgaggg atctccataa
     7621 gagaagaggg acagctatga ctgggagtag tcaggagagg aggaaaaatc tggctagtaa
     7681 aacatgtaag gaaaatttta gggatgttaa agaaaaaaat aacacaaaac aaaatat
//
