LOCUS       sequence-364154-        6814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optogenetic PHR domain coupled to GFP and FUSN domain with a high 
            phase separation propensity; binds to CIBN upon blue light exposure.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6814)
  AUTHORS   Trojanowski J, Frank L, Rademacher A, Mucke N, Grigaitis P, Rippe K
  TITLE     Transcription activation is enhanced by multivalent interactions 
            independent of phase separation.
  JOURNAL   Mol Cell. 2022 May 4. pii: S1097-2765(22)00328-8. doi: 
            10.1016/j.molcel.2022.04.017.
  PUBMED    35537448
REFERENCE   2  (bases 1 to 6814)
  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: "Mol Cell. 
            2022 May 4. pii: S1097-2765(22)00328-8. doi: 
            10.1016/j.molcel.2022.04.017."
COMMENT     Sequence Label: PHR-GFP-FUSN
FEATURES             Location/Qualifiers
     source          1..6814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..127
                     /gene="bla"
                     /label=AmpR promoter
     promoter        129..486
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        129..320
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(152..172)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      337..472
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             521..1315
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             521..1315
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(575..594)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1185..1204
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1503..1522)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1547..1594
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1923..2511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2412..2431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2686..2989
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2990..3193
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3144..3164
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3250..4743
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     CDS             join(4753..4755,4756..4758,4759..5469)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4753..4755,4756..4758,4759..5469)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4798..4819)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5059..5078)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5406..5427
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5850..5861
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    6227..6348
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6264..6283)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6318..6337
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6355..6810)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6492..6513)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6704..6723
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
       61 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
      121 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
      181 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
      241 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
      301 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
      361 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
      421 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
      481 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
      541 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
      601 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
      661 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
      721 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
      781 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
      841 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
      901 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
      961 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     1021 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     1081 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     1141 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     1201 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     1261 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     1321 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     1381 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     1441 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     1501 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     1561 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     1621 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     1681 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     1741 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     1801 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     1861 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     1921 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     1981 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2041 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     2101 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2161 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     2221 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2281 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     2341 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     2401 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     2461 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     2521 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     2581 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
     2641 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     2701 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     2761 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     2821 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     2881 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     2941 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     3001 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     3061 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     3121 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     3181 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga ctcagatctc
     3241 gaggccacca tgaagatgga caaaaagact atagtttggt ttagaagaga cctaaggatt
     3301 gaggataatc ctgcattagc agcagctgct cacgaaggat ctgtttttcc tgtcttcatt
     3361 tggtgtcctg aagaagaagg acagttttat cctggaagag cttcaagatg gtggatgaaa
     3421 caatcacttg ctcacttatc tcaatccttg aaggctcttg gatctgacct cactttaatc
     3481 aaaacccaca acacgatttc agcgatcttg gattgtatcc gcgttaccgg tgctacaaaa
     3541 gtcgtcttta accacctcta tgatcctgtt tcgttagttc gggaccatac cgtaaaggag
     3601 aagctggtgg aacgtgggat ctctgtgcaa agctacaatg gagatctatt gtatgaaccg
     3661 tgggagatat actgcgaaaa gggcaaacct tttacgagtt tcaattctta ctggaagaaa
     3721 tgcttagata tgtcgattga atccgttatg cttcctcctc cttggcggtt gatgccaata
     3781 actgcagcgg ctgaagcgat ttgggcgtgt tcgattgaag aactagggct ggagaatgag
     3841 gccgagaaac cgagcaatgc gttgttaact agagcttggt ctccaggatg gagcaatgct
     3901 gataagttac taaatgagtt catcgagaag cagttgatag attatgcaaa gaacagcaag
     3961 aaagttgttg ggaattctac ttcactactt tctccgtatc tccatttcgg ggaaataagc
     4021 gtcagacacg ttttccagtg tgcccggatg aaacaaatta tatgggcaag agataagaac
     4081 agtgaaggag aagaaagtgc agatcttttt cttaggggaa tcggtttaag agagtattct
     4141 cggtatatat gtttcaactt cccgtttact cacgagcaat cgttgttgag tcatcttcgg
     4201 tttttccctt gggatgctga tgttgataag ttcaaggcct ggagacaagg caggaccggt
     4261 tatccgttgg tggatgccgg aatgagagag ctttgggcta ccggatggat gcataacaga
     4321 ataagagtga ttgtttcaag ctttgctgtg aagtttcttc tccttccatg gaaatgggga
     4381 atgaagtatt tctgggatac acttttggat gctgatttgg aatgtgacat ccttggctgg
     4441 cagtatatct ctgggagtat ccccgatggc cacgagcttg atcgcttgga caatcccgcg
     4501 ttacaaggcg ccaaatatga cccagaaggt gagtacataa ggcaatggct tcccgagctt
     4561 gcgagattgc caactgaatg gatccatcat ccatgggacg ctcctttaac cgtactcaaa
     4621 gcttctggtg tggaactcgg aacaaactat gcgaaaccca ttgtagacat cgacacagct
     4681 cgtgagctac tagctaaagc tatttcaaga acccgtgaag cacagatcat gatcggagca
     4741 gcagcccggg tcatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     4801 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     4861 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     4921 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     4981 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     5041 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     5101 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     5161 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     5221 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     5281 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     5341 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     5401 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     5461 ctgtacaaga tggcctcaaa cgattatacc caacaagcaa cccaaagcta tggggcctac
     5521 cccacccagc ccgggcaggg ctattcccag cagagcagtc agccctacgg acagcagagt
     5581 tacagtggtt atagccagtc cacggacact tcaggctatg gccagagcag ctattcttct
     5641 tatggccaga gccagaacac aggctatgga actcagtcaa ctccccaggg atatggctcg
     5701 actggcggct atggcagtag ccagagctcc caatcgtctt acgggcagca gtcctcctat
     5761 cctggctatg gccagcagcc agctcccagc agcacctcgg gaagttacgg tagcagttct
     5821 cagagcagca gctatgggca gccccagagt gggagctaca gccagcagcc tagctatggt
     5881 ggacagcagc aaagctatgg acagcagcaa agctataatc cccctcaggg ctatggacag
     5941 cagaaccagt acaacagcag cagtggtggt ggaggtggag gtggaggtgg aggtaactat
     6001 ggccaagatc aatcctccat gagtagtggt ggtggcagtg gtggcggtta tggcaatcaa
     6061 gaccagagtg gtggaggtgg cagcggtggc tatggacagc aggaccgtgg atgacggcta
     6121 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     6181 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     6241 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6301 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg
     6361 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     6421 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     6481 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     6541 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     6601 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     6661 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     6721 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     6781 ccgcgcttaa tgcgccgcta cagggcgcgt cagg
//
