LOCUS       sequence-481814-        7219 bp    DNA     circular SYN 08-AUG-2025
DEFINITION  Blue-light-controlled release of gal4-vp16 transactivator from the 
            cytoplasm to the nucleus. Split version..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7219)
  TITLE     Photoswitchable intein for light control of covalent protein binding
            and cleavage
REFERENCE   2  (bases 1 to 7219)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: Split PS Intein gal4-vp16
FEATURES             Location/Qualifiers
     source          1..7219
                     /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             825..878
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             882..1817
                     /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             2424..2873
                     /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             2961..3398
                     /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             3588..3821
                     /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    3947..4068
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3984..4003)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4038..4057
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4075..4530)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4212..4233)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4424..4443
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4557..4661
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4663..5020
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4663..4854
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4686..4706)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4871..5006
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5055..5849
                     /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             5055..5849
                     /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(5109..5128)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5719..5738
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6037..6056)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6081..6128
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6457..7045
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6946..6965
                     /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 tgagcggcta ttgtctggat ctgaaaaccc aggttcagac accgcagggt atgaaagaaa
      601 tttcaaatat tcaggtgggt gatctggttc tgagcaatac tggttataat gaagtgctga
      661 atgtgtttcc gaaaagcaaa aaaaaaagct ataaaatcac cctggaagat ggcaaagaaa
      721 tcatttgtag cgaagaacac ctgtttccga cccagactgg tgaaatgaat attagcggtg
      781 gtctgaaaga aggtatgtgc ctgtatgtta aatccggaag cggcgagggc aggggcagcc
      841 tgctgacctg cggcgacgtg gaggagaacc ccggccccgg cgctggagac atgagagctg
      901 ccaacctttg gccaagcccg ctcatgatca aacgctctaa gaagaacagc ctggccttgt
      961 ccctgacggc cgaccagatg gtcagtgcct tgttggatgc tgagcccccc atactctatt
     1021 ccgagtatga tcctaccaga cccttcagtg aagcttcgat gatgggctta ctgaccaacc
     1081 tggcagacag ggagctggtt cacatgatca actgggcgaa gagggtgcca ggctttgtgg
     1141 atttgaccct ccatgatcag gtccaccttc tagaatgtgc ctggctagag atcctgatga
     1201 ttggtctcgt ctggcgctcc atggagcacc cagtgaagct actgtttgct cctaacttgc
     1261 tcttggacag gaaccaggga aaatgtgtag agggcatggt ggagatcttc gacatgctgc
     1321 tggctacatc atctcggttc cgcatgatga atctgcaggg agaggagttt gtgtgcctca
     1381 aatctattat tttgcttaat tctggagtgt acacatttct gtccagcacc ctgaagtctc
     1441 tggaagagaa ggaccatatc caccgagtcc tggacaagat cacagacact ttgatccacc
     1501 tgatggccaa ggcaggcctg accctgcagc agcagcacca gcggctggcc cagctcctcc
     1561 tcatcctctc ccacatcagg cacatgagta acaaaggcat ggagcatctg tacagcatga
     1621 agtgcaagaa cgtggtgccc ctctatgacc tgctgctgga ggcggcggac gcccaccgcc
     1681 tacatgcgcc cactagccgt ggaggggcat ccgtggagga gacggaccaa agccacttgg
     1741 ccactgcggg ctctacttca tcgcattcct tgcaaaagta ttacatcacg ggggaggcag
     1801 agggtttccc tgccacagct gggggtggtg gatcaaccgg tgaaatgatg ctgaaaaaaa
     1861 tcctgaaaat cgaggaactg gatgaacgcg gtaccggcag cggaggtcat accttgtatg
     1921 cgccgggggg ttatgacatc atgggttacc tcatacagat catgaatagg ccgaacccac
     1981 aagtggaact cggacccgtc gatacctcct gcgctctcat tctgtgtgac cttaagcaga
     2041 aggatacccc tatcgtgtac gcctccgagg catttctgta catgacaggg tactcgaacg
     2101 ccgaagtgct gggacggaac tgccgcttcc tgcaaagccc ggatggaatg gtgaagccta
     2161 agtcaacccg caaatacgtg gactccaaca ctatcaacac catgcgcaag gccattgacc
     2221 gcaatgctga ggtgcaagtg gaagtggtga acttcaagaa gaatggacag cgcttcgtca
     2281 acttcctgac tatgatcccc gtgcgcgacg agaccggcga atatcggtac agcatggggt
     2341 ttcagtgtga gacagagggc ggctcaggag gcagcggcgg ttctggaggt tccggtggcg
     2401 gctcaggagg tagcggaggg tctcacactc tttacgcccc tggaggatac gacattatgg
     2461 gatatttgat tcagattatg aaccgcccaa accctcaggt cgaactgggg cctgtggaca
     2521 cgtcatgtgc cctgatcctg tgcgatctga agcaaaagga cactccgatt gtctacgcct
     2581 cggaagcctt cttgtatatg accggataca gcaatgcaga ggtgctcggt aggaactgca
     2641 gattcctgca gtcccccgac gggatggtga aaccaaagtc cactcgcaaa tatgtggact
     2701 cgaacacgat caatacaatg cggaaggcca tcgaccggaa cgccgaggtc caggtggagg
     2761 tggtcaactt taagaagaac ggccagcggt tcgtgaactt tctcaccatg attccggtcc
     2821 gggatgaaac cggagagtac agatactcca tgggattcca gtgcgaaacc gaagggtcag
     2881 gatccgaact gattgatatt gaagttagcg gtaaccacct gttctatgcc aatgatattc
     2941 tgacccataa tagctcgagc aagctactgt cttctatcga acaagcatgc gatatttgcc
     3001 gacttaaaaa gctcaagtgc tccaaagaaa aaccgaagtg cgccaagtgt ctgaagaaca
     3061 actgggagtg tcgctactct cccaaaacca aaaggtctcc gctgactagg gcacatctga
     3121 cagaagtgga atcaaggcta gaaagactgg aacagctatt tctactgatt tttcctcgag
     3181 aagaccttga catgattttg aaaatggatt ctttacagga tataaaagca ttgttaacag
     3241 gattatttgt acaagataat gtgaataaag atgccgtcac agatagattg gcttcagtgg
     3301 agactgatat gcctctaaca ttgagacagc atagaataag tgcgacatca tcatcggaag
     3361 agagtagtaa caaaggtcaa agacagttga ctgtatcgcc ggaattcgga ggtggtggat
     3421 caggaggagg cggttctgcg tacagccgcg cgcgtacgaa aaacaattac gggtctacca
     3481 tcgagggcct gctcgatctc ccggacgacg acgcccccga agaggcgggg ctggcggctc
     3541 cgcgcctgtc ctttctcccc gcgggacaca cgcgcagact gtcgacggcc cccccgaccg
     3601 atgtcagcct gggggacgag ctccacttag acggcgagga cgtggcgatg gcgcatgccg
     3661 acgcgctaga cgatttcgat ctggacatgt tgggggacgg ggattcccct ggtccgggat
     3721 ttacccccca cgactccgcc ccctacggcg ctctggatat ggccgacttc gagtttgagc
     3781 agatgtttac cgatgccctt ggaattgacg agtacggtgg gtaaagcggc cgcgactcta
     3841 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     3901 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     3961 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4021 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg
     4081 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     4141 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     4201 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     4261 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     4321 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     4381 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     4441 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     4501 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     4561 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4621 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     4681 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4741 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4801 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4861 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4921 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     4981 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     5041 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5101 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     5161 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     5221 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5281 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5341 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5401 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5461 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5521 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5581 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5641 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5701 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5761 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5821 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5881 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5941 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     6001 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     6061 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     6121 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     6181 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     6241 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     6301 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6361 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6421 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6481 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6541 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6601 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6661 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6721 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6781 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6841 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6901 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6961 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7021 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7081 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     7141 ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata
     7201 gcccatatat ggagttccg
//
