LOCUS       sequence-364131-        6310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optogenetic PHR domain coupled to GFP; binds to CIBN upon blue light
            exposure.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6310)
  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 6310)
  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
FEATURES             Location/Qualifiers
     source          1..6310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..225
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     176..196
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             282..1775
                     /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(1785..1787,1788..1790,1791..2501)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1785..1787,1788..1790,1791..2501)
                     /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(1830..1851)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2091..2110)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2438..2459
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2535..2555
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2755..2876
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2792..2811)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2846..2865
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2883..3338)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3020..3041)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3232..3251
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3365..3469
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3471..3828
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3471..3662
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3494..3514)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3679..3814
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3863..4657
                     /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             3863..4657
                     /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(3917..3936)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4527..4546
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4845..4864)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4889..4936
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5265..5853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5754..5773
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(6028..6310,1..21)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
       61 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      121 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      181 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
      241 cagatccgct agcgctaccg gactcagatc tcgaggccac catgaagatg gacaaaaaga
      301 ctatagtttg gtttagaaga gacctaagga ttgaggataa tcctgcatta gcagcagctg
      361 ctcacgaagg atctgttttt cctgtcttca tttggtgtcc tgaagaagaa ggacagtttt
      421 atcctggaag agcttcaaga tggtggatga aacaatcact tgctcactta tctcaatcct
      481 tgaaggctct tggatctgac ctcactttaa tcaaaaccca caacacgatt tcagcgatct
      541 tggattgtat ccgcgttacc ggtgctacaa aagtcgtctt taaccacctc tatgatcctg
      601 tttcgttagt tcgggaccat accgtaaagg agaagctggt ggaacgtggg atctctgtgc
      661 aaagctacaa tggagatcta ttgtatgaac cgtgggagat atactgcgaa aagggcaaac
      721 cttttacgag tttcaattct tactggaaga aatgcttaga tatgtcgatt gaatccgtta
      781 tgcttcctcc tccttggcgg ttgatgccaa taactgcagc ggctgaagcg atttgggcgt
      841 gttcgattga agaactaggg ctggagaatg aggccgagaa accgagcaat gcgttgttaa
      901 ctagagcttg gtctccagga tggagcaatg ctgataagtt actaaatgag ttcatcgaga
      961 agcagttgat agattatgca aagaacagca agaaagttgt tgggaattct acttcactac
     1021 tttctccgta tctccatttc ggggaaataa gcgtcagaca cgttttccag tgtgcccgga
     1081 tgaaacaaat tatatgggca agagataaga acagtgaagg agaagaaagt gcagatcttt
     1141 ttcttagggg aatcggttta agagagtatt ctcggtatat atgtttcaac ttcccgttta
     1201 ctcacgagca atcgttgttg agtcatcttc ggtttttccc ttgggatgct gatgttgata
     1261 agttcaaggc ctggagacaa ggcaggaccg gttatccgtt ggtggatgcc ggaatgagag
     1321 agctttgggc taccggatgg atgcataaca gaataagagt gattgtttca agctttgctg
     1381 tgaagtttct tctccttcca tggaaatggg gaatgaagta tttctgggat acacttttgg
     1441 atgctgattt ggaatgtgac atccttggct ggcagtatat ctctgggagt atccccgatg
     1501 gccacgagct tgatcgcttg gacaatcccg cgttacaagg cgccaaatat gacccagaag
     1561 gtgagtacat aaggcaatgg cttcccgagc ttgcgagatt gccaactgaa tggatccatc
     1621 atccatggga cgctccttta accgtactca aagcttctgg tgtggaactc ggaacaaact
     1681 atgcgaaacc cattgtagac atcgacacag ctcgtgagct actagctaaa gctatttcaa
     1741 gaacccgtga agcacagatc atgatcggag cagcagcccg ggtcatggtg agcaagggcg
     1801 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     1861 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     1921 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     1981 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     2041 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2101 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2161 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2221 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     2281 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     2341 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     2401 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     2461 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gagcggccgc agggggcgtc
     2521 gactgggtac ccctcctaag aagaagagga aggttgcccc caccatggat aaaaagaaga
     2581 gaaaggcccc cccgaccgat gtcagcctgg gggacggtcc ggatctagac atctcgatct
     2641 agcggctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     2701 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     2761 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2821 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     2881 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     2941 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3001 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3061 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3121 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3181 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3241 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3301 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3361 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3421 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     3481 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3541 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3601 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3661 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3721 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3781 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     3841 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3901 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3961 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4021 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     4081 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4141 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4201 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4261 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4321 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4381 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4441 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4501 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4561 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4621 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4681 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4741 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4801 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     4861 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     4921 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4981 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     5041 ccaagttcgg gtgaaggcct agggctcgca gccaacgtcg gggcggcagg ccctgccata
     5101 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5161 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5221 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5281 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5341 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5401 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5461 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5521 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5581 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5641 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5701 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5761 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5821 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5881 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5941 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
     6001 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     6061 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     6121 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     6181 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     6241 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     6301 catctacgta
//
