LOCUS       sequence-116162-        6650 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Blue light dependent photoswitching system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6650)
  AUTHORS   Kawano F, Suzuki H, Furuya A, Sato M
  TITLE     Engineered pairs of distinct photoswitches for optogenetic control 
            of cellular proteins.
  JOURNAL   Nat Commun. 2015 Feb 24;6:6256. doi: 10.1038/ncomms7256.
  PUBMED    25708714
REFERENCE   2  (bases 1 to 6650)
  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: "2015-02-24"; volume: "6:6256. doi"; pages: " 
            10.1038/ncomms7256"
COMMENT     Sequence Label: pcDNA3.1_nMagHigh1-EGFP-CAAX
FEATURES             Location/Qualifiers
     source          1..6650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      932..941
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             944..1393
                     /codon_start=1
                     /product="""negative Magnet"" variant of an N-terminally 
                     truncated Vivid (VVD) blue light photoreceptor from 
                     Neurospora crassa (Kawano et al., 2015)"
                     /label=nMagHigh1
                     /note="rapidly heterodimerizes with pMag or pMagFast2 upon 
                     blue light irradiation"
                     /translation="HTLYAPGGYDIMGYLDQIGNRPNPQVELGPVDTSCALILCDLKQK
                     DTPIVYASEAFLYMTGYSNAEVLGRNCRFLQSPDGMVKPKSTRKYVDSNTINTIRKAID
                     RNAEVQVEVVNFKKNGQRFVNFLTIIPVRDETGEYRYSMGFQCETE"
     CDS             join(1442..1444,1445..1447,1448..2158)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2244..2261)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2250..2474
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2520..2948
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2607..2626)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2817..2838
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2957..2977)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2962..3291
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3142..3277
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3204..3223
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3358..4152
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3412..3431)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4022..4041
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4326..4447
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4363..4382)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4417..4436
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4496..4512)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4496..4512)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4509..4531)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4520..4536
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4544..4550,4551..4568,4569..4574))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4589..4610
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4727..4744)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4898..5483)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4978..4997)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5654..6445,6446..6514))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6277..6296
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6515..6619)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttaagc tggctagcgc cgccaccatg ggacacactc tttacgcccc
      961 tggaggatac gacattatgg gatatttgga tcagattggg aaccgcccaa accctcaggt
     1021 cgaactgggg cctgtggaca cgtcatgtgc cctgatcctg tgcgatctga agcaaaagga
     1081 cactccgatc gtctacgcct cggaagcctt cttgtatatg accggataca gcaatgcaga
     1141 ggtgctcggc aggaactgca gattcctgca gtcccccgac gggatggtga aaccaaagtc
     1201 gactcgcaaa tatgtggact cgaacacgat caacaccatc cggaaggcca tcgaccggaa
     1261 cgccgaggtc caggtggagg tggtcaactt taagaagaac ggccagcggt tcgtgaactt
     1321 tctgaccatc attccggtcc gggatgaaac cggagagtac agatactcca tgggattcca
     1381 gtgcgaaacc gaaggcggct ccggaggcag cggcggttct ggaggttccg gtggcggctc
     1441 catggtgtcg aagggagagg agcttttcac cggcgtggtg ccgattcttg tggaactgga
     1501 cggcgacgtg aacggacata agttcagcgt gtcaggagaa ggagaagggg acgcgactta
     1561 cggaaagctg accctgaagt tcatctgtac caccgggaag ctcccggtgc cctggccgac
     1621 cctcgtgact actctgacct acggagtgca gtgcttctcc cgctacccgg accatatgaa
     1681 gcagcacgat ttcttcaaga gcgctatgcc cgagggttac gtgcaagaac ggaccatctt
     1741 cttcaaggac gacggaaact acaagaccag agccgaagtg aaattcgaag gagacaccct
     1801 cgtcaatcgc atcgagctga agggaatcga ctttaaggaa gatgggaata tcctgggtca
     1861 caagctggag tacaactaca actcccacaa tgtgtacatc atggcggata agcaaaagaa
     1921 cggcattaag gtcaacttca agatccggca taacatagaa gatggctccg tgcaactggc
     1981 ggaccactac cagcagaaca cccccattgg ggacggtccg gtgctgctcc ctgataacca
     2041 ctacctgtca acccaaagcg cactgtccaa ggaccccaac gagaagaggg accacatggt
     2101 gctgctggag ttcgtcactg ccgctggcat cactctcgga atggacgagc tgtacaaggg
     2161 gaagaaaaag aagaagaagt ccaagacaaa atgcgtgatt atgtagctcg agtctagagg
     2221 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     2281 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2341 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     2401 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     2461 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     2521 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2581 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2641 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2701 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2761 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2821 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2881 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2941 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3001 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3061 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3121 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3181 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3241 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     3301 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     3361 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3421 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3481 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3541 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3601 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3661 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3721 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3781 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     3841 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     3901 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     3961 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4021 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4081 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4141 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4201 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4261 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4321 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4381 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4441 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     4501 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4561 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4621 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4681 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4741 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4801 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4861 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4921 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4981 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5041 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5101 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5161 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5221 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5281 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5341 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5401 cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta
     5461 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5521 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5581 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5641 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5701 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5761 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5821 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5881 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5941 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6001 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6061 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6121 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6181 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6241 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6301 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6361 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6421 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6481 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6541 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6601 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
