LOCUS       sequence-481860-        8428 bp    DNA     circular SYN 08-AUG-2025
DEFINITION  Blue-light-controlled release of tTA transactivator from the 
            cytoplasm to the nucleus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8428)
  TITLE     Photoswitchable intein for light control of covalent protein binding
            and cleavage
REFERENCE   2  (bases 1 to 8428)
  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: Cp PS Intein tTA
FEATURES             Location/Qualifiers
     source          1..8428
                     /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             join(2667..3284,3285..3668)
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus 
                     VP16"
                     /label=tTA
                     /note="In the Tet-Off(R) system, tTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="SRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVK
                     NKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRP
                     TEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTD
                     SMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTIE
                     GLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHAD
                     ALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(2737..2756)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     polyA_signal    5156..5277
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5193..5212)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5247..5266
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5284..5739)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5421..5442)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5633..5652
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5766..5870
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5872..6229
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5872..6063
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5895..5915)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6080..6215
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6264..7058
                     /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             6264..7058
                     /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(6318..6337)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6928..6947
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7246..7265)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7290..7337
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7666..8254
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8155..8174
                     /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 tcgagctcta gattagataa aagtaaagtg attaacagcg
     2701 cattagagct gcttaatgag gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga
     2761 agctaggtgt agagcagcct acattgtatt ggcatgtaaa aaataagcgg gctttgctcg
     2821 acgccttagc cattgagatg ttagataggc accatactca cttttgccct ttagaagggg
     2881 aaagctggca agatttttta cgtaataacg ctaaaagttt tagatgtgct ttactaagtc
     2941 atcgcgatgg agcaaaagta catttaggta cacggcctac agaaaaacag tatgaaactc
     3001 tcgaaaatca attagccttt ttatgccaac aaggtttttc actagagaat gcattatatg
     3061 cactcagcgc tgtggggcat tttactttag gttgcgtatt ggaagatcaa gagcatcaag
     3121 tcgctaaaga agaaagggaa acacctacta ctgatagtat gccgccatta ttacgacaag
     3181 ctatcgaatt atttgatcac caaggtgcag agccagcctt cttattcggc cttgaattga
     3241 tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg gtccgcgtac agccgcgcgc
     3301 gtacgaaaaa caattacggg tctaccatcg agggcctgct cgatctcccg gacgacgacg
     3361 cccccgaaga ggcggggctg gcggctccgc gcctgtcctt tctccccgcg ggacacacgc
     3421 gcagactgtc gacggccccc ccgaccgatg tcagcctggg ggacgagctc cacttagacg
     3481 gcgaggacgt ggcgatggcg catgccgacg cgctagacga tttcgatctg gacatgttgg
     3541 gggacgggga ttccccgggt ccgggattta ccccccacga ctccgccccc tacggcgctc
     3601 tggatatggc cgacttcgag tttgagcaga tgtttaccga tgcccttgga attgacgagt
     3661 acggtgggac tagtggaggg tcaggggctg gagggtcggg ggcaggcggt tcgggagccg
     3721 gaggatctgg agggggtggt tcgggtggtg gtggtagcgg ctactgcctg gacctgaaga
     3781 cccaggtgca gaccccccag ggcatgaagg agatcagcaa catccaggtg ggcgacctgg
     3841 tgctgagcaa caccggctac aacgaggtgc tgaacgtgtt ccccaagagc aagaagaaga
     3901 gctacaagat caccctggag gacggcaagg agatcatctg cagcgaggag cacctgttcc
     3961 ccacccagac cggcgagatg aacatcagcg gcggcctgaa ggagggcatg tgcctgtacg
     4021 tgaagggcgg ctccggagga ggtagtggcg gagggagtgg tgggagtggc tctggttcag
     4081 gcagtggttc tgcaggcgat atgcgggccg ctaatttgtg gccttctcct ttgatgatta
     4141 agagaagtaa aaaaaattcc ttggctctga gcttgaccgc tgatcaaatg gtgtccgctc
     4201 tcctcgacgc cgaacctcct atcttgtaca gcgaatacga ccccacacgg ccatttagcg
     4261 aggcctcaat gatgggactg cttacaaatc tcgctgatag agaactcgtg catatgatta
     4321 attgggccaa acgggtcccc ggattcgtcg acctgactct gcatgaccaa gtgcatctct
     4381 tggagtgcgc atggctggaa attctcatga tcggcctggt gtggcggagc atggaacatc
     4441 ctgtcaaact gctcttcgcc cccaatctgc tgctggatcg gaatcaaggc aagtgcgtgg
     4501 aaggaatggt cgaaattttt gatatgctcc tcgccaccag cagcaggttt agaatgatga
     4561 acctccaagg cgaagagttc gtctgtctga agagcatcat cctgctgaac agcggggtct
     4621 ataccttcct aagtagtaca ttaaaaagtt tggaggaaaa agatcacatt catcgggtgc
     4681 tcgataaaat taccgatacc ctaattcatc tcatggctaa agccggactc acactccaac
     4741 aacaacatca aagactcgct caactgttac tgattctgag ccatattaga catatgagca
     4801 ataagggaat ggaacacctc tattccatga aatgtaaaaa tgtcgtccct ctgtacgatc
     4861 tcctcctcga agccgccgat gctcataggc tgcacgcccc tacctccaga ggcggagcca
     4921 gcgtcgaaga aaccgatcag tcccatctgg ctaccgccgg aagcacctcc agccacagcc
     4981 ttcagaaata ctatattacc ggcgaagccg aaggctttcc cgctaccgcc taaagcggcc
     5041 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     5101 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     5161 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5221 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     5281 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     5341 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     5401 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     5461 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     5521 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     5581 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     5641 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     5701 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     5761 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5821 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     5881 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     5941 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     6001 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     6061 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     6121 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     6181 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     6241 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     6301 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     6361 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     6421 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     6481 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     6541 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     6601 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     6661 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     6721 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     6781 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     6841 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     6901 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     6961 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     7021 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     7081 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     7141 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     7201 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     7261 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     7321 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     7381 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     7441 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     7501 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     7561 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7621 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     7681 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     7741 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     7801 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     7861 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     7921 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     7981 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     8041 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     8101 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     8161 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     8221 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     8281 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     8341 tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat
     8401 tagttcatag cccatatatg gagttccg
//
