LOCUS       sequence-483143-        7240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Blue-light-induced reconstitution of the tTA transactivator via 
            split PS Intein-mediated fusion of TetR and vp16..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7240)
  AUTHORS   Baloban M, Manoilov KY, Karasev MM, Verkhusha VV, Shcherbakova DM
  TITLE     Photoswitchable intein for light control of covalent protein binding
            and cleavage.
  JOURNAL   Nat Commun. 2025 Sep 11;16(1):8263. doi: 10.1038/s41467-025-63595-9.
  PUBMED    40935816
REFERENCE   2  (bases 1 to 7240)
  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: "Nat 
            Commun."; date: "2025-09-11"; volume: "16(1):8263. doi"; pages: " 
            10.1038/s41467-025-63595-9"
COMMENT     Sequence Label: Split PS Intein TetR-vp16
FEATURES             Location/Qualifiers
     source          1..7240
                     /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             540..1160
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(613..632)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             1467..1520
                     /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             2106..2555
                     /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             2823..3056
                     /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"
     CDS             3072..3125
                     /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             join(3126..3128,3129..3131,3132..3845)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3171..3192)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3432..3451)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3779..3800
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3968..4089
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4005..4024)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4059..4078
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4096..4551)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4233..4254)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4445..4464
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4578..4682
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4684..5041
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4684..4875
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4707..4727)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4892..5027
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5076..5870
                     /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             5076..5870
                     /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(5130..5149)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5740..5759
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6058..6077)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6102..6149
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6478..7066
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6967..6986
                     /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 tgtctagatt agataaaagt aaagtgatta acagcgcatt agagctgctt aatgaggtcg
      601 gaatcgaagg tttaacaacc cgtaaactcg cccagaagct aggtgtagag cagcctacat
      661 tgtattggca tgtaaaaaat aagcgggctt tgctcgacgc cttagccatt gagatgttag
      721 ataggcacca tactcacttt tgccctttag aaggggaaag ctggcaagat tttttacgta
      781 ataacgctaa aagttttaga tgtgctttac taagtcatcg cgatggagca aaagtacatt
      841 taggtacacg gcctacagaa aaacagtatg aaactctcga aaatcaatta gcctttttat
      901 gccaacaagg tttttcacta gagaatgcat tatatgcact cagcgctgtg gggcatttta
      961 ctttaggttg cgtattggaa gatcaagagc atcaagtcgc taaagaagaa agggaaacac
     1021 ctactactga tagtatgccg ccattattac gacaagctat cgaattattt gatcaccaag
     1081 gtgcagagcc agccttctta ttcggccttg aattgatcat atgcggatta gaaaaacaac
     1141 ttaaatgtga aagtgggtcc gggggtggag ggtcgggtgg aggaggttcc ggatactgtc
     1201 tggatctgaa aacccaggtt cagacaccgc agggtatgaa agaaatttca aatattcagg
     1261 tgggtgatct ggttctgagc aatactggtt ataatgaagt gctgaatgtg tttccgaaaa
     1321 gcaaaaaaaa aagctataaa atcaccctgg aagatggcaa agaaatcatt tgtagcgaag
     1381 aacacctgtt tccgacccag actggtgaaa tgaatattag cggtggtctg aaagaaggta
     1441 tgtgcctgta tgttaaaggc agcggcgagg gcaggggcag cctgctgacc tgcggcgacg
     1501 tggaggagaa ccccggcccc ggcgaaatga tgctgaaaaa aatcctgaaa atcgaggaac
     1561 tggatgaacg cggtaccggc agcggaggtc ataccttgta tgcgccgggg ggttatgaca
     1621 tcatgggtta cctcatacag atcatgaata ggccgaaccc acaagtggaa ctcggacccg
     1681 tcgatacctc ctgcgctctc attctgtgtg accttaagca gaaggatacc cctatcgtgt
     1741 acgcctccga ggcatttctg tacatgacag ggtactcgaa cgccgaagtg ctgggacgga
     1801 actgccgctt cctgcaaagc ccggatggaa tggtgaagcc taagtcaacc cgcaaatacg
     1861 tggactccaa cactatcaac accatgcgca aggccattga ccgcaatgct gaggtgcaag
     1921 tggaagtggt gaacttcaag aagaatggac agcgcttcgt caacttcctg actatgatcc
     1981 ccgtgcgcga cgagaccggc gaatatcggt acagcatggg gtttcagtgt gagacagagg
     2041 gcggctcagg aggcagcggc ggttctggag gttccggtgg cggctcagga ggtagcggag
     2101 ggtctcacac tctttacgcc cctggaggat acgacattat gggatatttg attcagatta
     2161 tgaaccgccc aaaccctcag gtcgaactgg ggcctgtgga cacgtcatgt gccctgatcc
     2221 tgtgcgatct gaagcaaaag gacactccga ttgtctacgc ctcggaagcc ttcttgtata
     2281 tgaccggata cagcaatgca gaggtgctcg gtaggaactg cagattcctg cagtcccccg
     2341 acgggatggt gaaaccaaag tccactcgca aatatgtgga ctcgaacacg atcaatacaa
     2401 tgcggaaggc catcgaccgg aacgccgagg tccaggtgga ggtggtcaac tttaagaaga
     2461 acggccagcg gttcgtgaac tttctcacca tgattccggt ccgggatgaa accggagagt
     2521 acagatactc catgggattc cagtgcgaaa ccgaagggtc aggatccgaa ctgattgata
     2581 ttgaagttag cggtaaccac ctgttctatg ccaatgatat tctgacccat aatagctcga
     2641 gcggaggggg aggatctgga gggggtggtt cggcgtacag ccgcgcgcgt acgaaaaaca
     2701 attacgggtc taccatcgag ggcctgctcg atctcccgga cgacgacgcc cccgaagagg
     2761 cggggctggc ggctccgcgc ctgtcctttc tccccgcggg acacacgcgc agactgtcga
     2821 cggccccccc gaccgatgtc agcctggggg acgagctcca cttagacggc gaggacgtgg
     2881 cgatggcgca tgccgacgcg ctagacgatt tcgatctgga catgttgggg gacggggatt
     2941 ccccgggtcc gggatttacc ccccacgact ccgcccccta cggcgctctg gatatggccg
     3001 acttcgagtt tgagcagatg tttaccgatg cccttggaat tgacgagtac ggtggggaat
     3061 tcggtagcgg cgagggcaga ggaagtctgc taacatgcgg tgatgtcgag gagaatcctg
     3121 gcccaatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     3181 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     3241 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     3301 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     3361 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     3421 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     3481 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     3541 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     3601 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     3661 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3721 accactacct gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca
     3781 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3841 agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     3901 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     3961 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4021 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4081 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     4141 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     4201 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4261 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4321 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4381 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4441 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     4501 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     4561 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4621 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4681 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4741 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4801 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4861 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4921 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4981 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     5041 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     5101 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     5161 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     5221 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     5281 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5341 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5401 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5461 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5521 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5581 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     5641 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5701 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5761 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5821 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5881 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5941 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     6001 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     6061 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     6121 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     6181 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     6241 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     6301 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     6361 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6421 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6481 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6541 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6601 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6661 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6721 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6781 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6841 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6901 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6961 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7021 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7081 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     7141 atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa
     7201 ttacggggtc attagttcat agcccatata tggagttccg
//
