LOCUS       sequence-483210-        8287 bp    DNA     circular SYN 14-AUG-2025
DEFINITION  Blue-light-controlled release of gal4-vp16 transactivator from the 
            cytoplasm to the nucleus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8287)
  TITLE     Photoswitchable intein for light control of covalent protein binding
            and cleavage
REFERENCE   2  (bases 1 to 8287)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: Cp PS Intein gal4-vp16
FEATURES             Location/Qualifiers
     source          1..8287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             543..1478
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             2130..2579
                     /codon_start=1
                     /product="""positive Magnet"" variant of an N-terminally 
                     truncated Vivid (VVD) blue light photoreceptor from 
                     Neurospora crassa (Kawano et al., 2015)"
                     /label=pMag
                     /note="rapidly heterodimerizes with nMagHigh1 upon blue 
                     light irradiation to form a complex with slow switch-off 
                     kinetics"
                     /translation="HTLYAPGGYDIMGYLIQIMNRPNPQVELGPVDTSCALILCDLKQK
                     DTPIVYASEAFLYMTGYSNAEVLGRNCRFLQSPDGMVKPKSTRKYVDSNTINTMRKAID
                     RNAEVQVEVVNFKKNGQRFVNFLTMIPVRDETGEYRYSMGFQCETE"
     CDS             2667..3104
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="KLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKR
                     SPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKD
                     AVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             3294..3527
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     polyA_signal    5015..5136
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5052..5071)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5106..5125
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5143..5598)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5280..5301)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5492..5511
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5625..5729
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5731..6088
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5731..5922
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5754..5774)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5939..6074
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6123..6917
                     /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             6123..6917
                     /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(6177..6196)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6787..6806
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7105..7124)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7149..7196
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7525..8113
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8014..8033
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcacca
      541 tggctggaga catgagagct gccaaccttt ggccaagccc gctcatgatc aaacgctcta
      601 agaagaacag cctggccttg tccctgacgg ccgaccagat ggtcagtgcc ttgttggatg
      661 ctgagccccc catactctat tccgagtatg atcctaccag acccttcagt gaagcttcga
      721 tgatgggctt actgaccaac ctggcagaca gggagctggt tcacatgatc aactgggcga
      781 agagggtgcc aggctttgtg gatttgaccc tccatgatca ggtccacctt ctagaatgtg
      841 cctggctaga gatcctgatg attggtctcg tctggcgctc catggagcac ccagtgaagc
      901 tactgtttgc tcctaacttg ctcttggaca ggaaccaggg aaaatgtgta gagggcatgg
      961 tggagatctt cgacatgctg ctggctacat catctcggtt ccgcatgatg aatctgcagg
     1021 gagaggagtt tgtgtgcctc aaatctatta ttttgcttaa ttctggagtg tacacatttc
     1081 tgtccagcac cctgaagtct ctggaagaga aggaccatat ccaccgagtc ctggacaaga
     1141 tcacagacac tttgatccac ctgatggcca aggcaggcct gaccctgcag cagcagcacc
     1201 agcggctggc ccagctcctc ctcatcctct cccacatcag gcacatgagt aacaaaggca
     1261 tggagcatct gtacagcatg aagtgcaaga acgtggtgcc cctctatgac ctgctgctgg
     1321 aggcggcgga cgcccaccgc ctacatgcgc ccactagccg tggaggggca tccgtggagg
     1381 agacggacca aagccacttg gccactgcgg gctctacttc atcgcattcc ttgcaaaagt
     1441 attacatcac gggggaggca gagggtttcc ctgccacagc tgggggtggt ggatcaggag
     1501 gtggtggatc aggaggaggt ggttctggtg gaggaggttc taccggtgaa atgatgctga
     1561 aaaaaatcct gaaaatcgag gaactggatg aacgcggtac cggcagcgga ggtcatacct
     1621 tgtatgcgcc ggggggttat gacatcatgg gttacctcat acagatcatg aataggccga
     1681 acccacaagt ggaactcgga cccgtcgata cctcctgcgc tctcattctg tgtgacctta
     1741 agcagaagga tacccctatc gtgtacgcct ccgaggcatt tctgtacatg acagggtact
     1801 cgaacgccga agtgctggga cggaactgcc gcttcctgca aagcccggat ggaatggtga
     1861 agcctaagtc aacccgcaaa tacgtggact ccaacactat caacaccatg cgcaaggcca
     1921 ttgaccgcaa tgctgaggtg caagtggaag tggtgaactt caagaagaat ggacagcgct
     1981 tcgtcaactt cctgactatg atccccgtgc gcgacgagac cggcgaatat cggtacagca
     2041 tggggtttca gtgtgagaca gagggcggct caggaggcag cggcggttct ggaggttccg
     2101 gtggcggctc aggaggtagc ggagggtctc acactcttta cgcccctgga ggatacgaca
     2161 ttatgggata tttgattcag attatgaacc gcccaaaccc tcaggtcgaa ctggggcctg
     2221 tggacacgtc atgtgccctg atcctgtgcg atctgaagca aaaggacact ccgattgtct
     2281 acgcctcgga agccttcttg tatatgaccg gatacagcaa tgcagaggtg ctcggtagga
     2341 actgcagatt cctgcagtcc cccgacggga tggtgaaacc aaagtccact cgcaaatatg
     2401 tggactcgaa cacgatcaat acaatgcgga aggccatcga ccggaacgcc gaggtccagg
     2461 tggaggtggt caactttaag aagaacggcc agcggttcgt gaactttctc accatgattc
     2521 cggtccggga tgaaaccgga gagtacagat actccatggg attccagtgc gaaaccgaag
     2581 ggtcaggatc cgaactgatt gatattgaag ttagcggtaa ccacctgttc tatgccaatg
     2641 atattctgac ccataatagc tcgagcaagc tactgtcttc tatcgaacaa gcatgcgata
     2701 tttgccgact taaaaagctc aagtgctcca aagaaaaacc gaagtgcgcc aagtgtctga
     2761 agaacaactg ggagtgtcgc tactctccca aaaccaaaag gtctccgctg actagggcac
     2821 atctgacaga agtggaatca aggctagaaa gactggaaca gctatttcta ctgatttttc
     2881 ctcgagaaga ccttgacatg attttgaaaa tggattcttt acaggatata aaagcattgt
     2941 taacaggatt atttgtacaa gataatgtga ataaagatgc cgtcacagat agattggctt
     3001 cagtggagac tgatatgcct ctaacattga gacagcatag aataagtgcg acatcatcat
     3061 cggaagagag tagtaacaaa ggtcaaagac agttgactgt atcgccggaa ttcggaggtg
     3121 gtggatcagg aggaggcggt tctgcgtaca gccgcgcgcg tacgaaaaac aattacgggt
     3181 ctaccatcga gggcctgctc gatctcccgg acgacgacgc ccccgaagag gcggggctgg
     3241 cggctccgcg cctgtccttt ctccccgcgg gacacacgcg cagactgtcg acggcccccc
     3301 cgaccgatgt cagcctgggg gacgagctcc acttagacgg cgaggacgtg gcgatggcgc
     3361 atgccgacgc gctagacgat ttcgatctgg acatgttggg ggacggggat tcccctggtc
     3421 cgggatttac cccccacgac tccgccccct acggcgctct ggatatggcc gacttcgagt
     3481 ttgagcagat gtttaccgat gcccttggaa ttgacgagta cggtgggact agtggagggt
     3541 caggggctgg agggtcgggg gcaggcggtt cgggagccgg aggatctgga gggggtggtt
     3601 cgggtggtgg tggtagcggc tactgcctgg acctgaagac ccaggtgcag accccccagg
     3661 gcatgaagga gatcagcaac atccaggtgg gcgacctggt gctgagcaac accggctaca
     3721 acgaggtgct gaacgtgttc cccaagagca agaagaagag ctacaagatc accctggagg
     3781 acggcaagga gatcatctgc agcgaggagc acctgttccc cacccagacc ggcgagatga
     3841 acatcagcgg cggcctgaag gagggcatgt gcctgtacgt gaagggcggc tccggaggag
     3901 gtagtggcgg agggagtggt gggagtggct ctggttcagg cagtggttct gcaggcgata
     3961 tgcgggccgc taatttgtgg ccttctcctt tgatgattaa gagaagtaaa aaaaattcct
     4021 tggctctgag cttgaccgct gatcaaatgg tgtccgctct cctcgacgcc gaacctccta
     4081 tcttgtacag cgaatacgac cccacacggc catttagcga ggcctcaatg atgggactgc
     4141 ttacaaatct cgctgataga gaactcgtgc atatgattaa ttgggccaaa cgggtccccg
     4201 gattcgtcga cctgactctg catgaccaag tgcatctctt ggagtgcgca tggctggaaa
     4261 ttctcatgat cggcctggtg tggcggagca tggaacatcc tgtcaaactg ctcttcgccc
     4321 ccaatctgct gctggatcgg aatcaaggca agtgcgtgga aggaatggtc gaaatttttg
     4381 atatgctcct cgccaccagc agcaggttta gaatgatgaa cctccaaggc gaagagttcg
     4441 tctgtctgaa gagcatcatc ctgctgaaca gcggggtcta taccttccta agtagtacat
     4501 taaaaagttt ggaggaaaaa gatcacattc atcgggtgct cgataaaatt accgataccc
     4561 taattcatct catggctaaa gccggactca cactccaaca acaacatcaa agactcgctc
     4621 aactgttact gattctgagc catattagac atatgagcaa taagggaatg gaacacctct
     4681 attccatgaa atgtaaaaat gtcgtccctc tgtacgatct cctcctcgaa gccgccgatg
     4741 ctcataggct gcacgcccct acctccagag gcggagccag cgtcgaagaa accgatcagt
     4801 cccatctggc taccgccgga agcacctcca gccacagcct tcagaaatac tatattaccg
     4861 gcgaagccga aggctttccc gctaccgcct aaagcggccg cgactctaga tcataatcag
     4921 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     4981 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     5041 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5101 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     5161 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     5221 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     5281 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     5341 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     5401 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     5461 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     5521 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     5581 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5641 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5701 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     5761 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     5821 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     5881 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     5941 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     6001 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     6061 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     6121 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     6181 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     6241 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     6301 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     6361 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     6421 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     6481 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     6541 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     6601 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     6661 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     6721 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     6781 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     6841 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     6901 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     6961 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     7021 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     7081 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     7141 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     7201 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     7261 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     7321 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     7381 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     7441 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     7501 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     7561 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     7621 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     7681 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     7741 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     7801 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     7861 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     7921 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     7981 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     8041 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     8101 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     8161 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     8221 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     8281 agttccg
//
