LOCUS       sequence-116158-        8450 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 8450)
  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 8450)
  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_iSH2-pMagFast2(3x)-iRFP
FEATURES             Location/Qualifiers
     source          1..8450
                     /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      917..926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1568..2017
                     /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=pMagFast2
                     /note="rapidly heterodimerizes with nMagHigh1 upon blue 
                     light irradiation to form a complex with fast switch-off 
                     kinetics"
                     /translation="HTLYAPGGYDIMGYLRQIRNRPNPQVELGPVDTSCALVLCDLKQK
                     DTPVVYASEAFLYMTGYSNAEVLGRNCRFLQSPDGMVKPKSTRKYVDSNTINTMRKAID
                     RNAEVQVEVVNFKKNGQRFVNFLTMIPVRDETGEYRYSMGFQCETE"
     primer_bind     complement(4044..4061)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4050..4274
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4320..4748
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4407..4426)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4617..4638
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4757..4777)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4762..5091
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4942..5077
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5004..5023
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5158..5952
                     /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(5212..5231)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5822..5841
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6126..6247
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6163..6182)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6217..6236
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6296..6312)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6296..6312)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6309..6331)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6320..6336
                     /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(6344..6350,6351..6368,6369..6374))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6389..6410
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6527..6544)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6698..7283)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6778..6797)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7454..8245,8246..8314))
                     /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     8077..8096
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8315..8419)
                     /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 aagcttgcca ccatggtgtc caaataccaa caggatcaag ttgtcaaaga
      961 agataatatt gaagctgtag ggaaaaaatt acatgaatat aacactcaat ttcaagaaaa
     1021 aagtcgggaa tatgatagat tatatgaaga ctatacccga acttcccagg aaatccaaat
     1081 gaaaagaaca gctattgaag catttaatga aaccataaaa atatttgaag aacagtgcca
     1141 aacccaggag cggtacagca aagaatacat agaaaagttc aaacgtgaag gcaacgagac
     1201 ggaaatccaa aggattatgc ataattatga aaagttaaag tctcgaatca gtgaaattgt
     1261 cgacagcaga aggagattgg aagaggactt gaagaagcag gcggctgagt atcgtgagat
     1321 tgacaaacgc atgaacagca ttaaaccaga cctcattcag ctaagaaaga cgagagacca
     1381 atacttgatg tggttgactc agaaaggtgt tcggcagaag aaattgaacg agtggctggg
     1441 caatgaaaac actgaagacc aatattcgct ggtagaagat gatgaggatt tgccccacca
     1501 cgatgagaag actggatccg gcggctccgg aggcagcggc ggttctggag gttccggtgg
     1561 cggctcccac actctttacg cccctggagg atacgacatt atgggatatt tgcggcagat
     1621 taggaaccgc ccaaaccctc aggtcgaact ggggcctgtg gacacgtcat gtgccctggt
     1681 cctgtgcgat ctgaagcaaa aggacactcc ggtggtctac gcctcggaag ccttcttgta
     1741 tatgaccgga tacagcaatg cagaggtgct cggcaggaac tgcagattcc tgcagtcccc
     1801 cgacgggatg gtgaaaccaa agtcgactcg caaatatgtg gactcgaaca cgatcaacac
     1861 catgcggaag gccatcgacc ggaacgccga ggtccaggtg gaggtggtca actttaagaa
     1921 gaacggccag cggttcgtga actttctgac catgattccg gtccgggatg aaaccggaga
     1981 gtacagatac tccatgggat tccagtgcga aaccgaaggt tccggcggct cgggcggatc
     2041 agggggaagc ggacacacac tctacgcccc gggaggctac gatatcatgg gctacctgag
     2101 gcagatccgg aacagaccga acccccaagt ggaactcggc cccgtggaca cttcttgcgc
     2161 tctggtgctg tgcgacctga agcagaagga tacccccgtg gtgtacgcat ccgaagcgtt
     2221 cctctacatg accgggtact cgaacgcgga ggtgctgggt cgcaactgcc ggttcctgca
     2281 gagccccgac ggaatggtca agcccaagtc cactcggaaa tacgtggatt ccaacaccat
     2341 caatactatg aggaaggcta tcgataggaa cgcggaagtc caagtggagg tggtgaactt
     2401 caagaagaat ggccagagat tcgtcaattt ccttaccatg atccctgtgc gcgatgagac
     2461 tggagagtac cgatactcaa tgggcttcca gtgtgaaacc gagggaagcg gcggttccgg
     2521 ggggtcggga ggttccggac ataccctcta cgcgccgggg ggttatgaca tcatgggtta
     2581 cctcagacag atcagaaacc ggccgaaccc acaagtggag ctgggacccg tcgacacctc
     2641 ctgcgccctc gtgctgtgtg accttaagca gaaggacacc cctgtggtgt acgcctccga
     2701 agcattcctg tacatgaccg ggtactcgaa cgccgaagtg ctgggacgga actgccgctt
     2761 cctgcaatcc ccggatggaa tggtgaagcc taagtcaacc cgcaaatacg tggactccaa
     2821 cactatcaac accatgcgca aggccattga ccgcaatgct gaggtgcaag tggaagtggt
     2881 gaacttcaag aagaacggac agcgcttcgt caacttcctg actatgattc ccgtgcggga
     2941 cgaaaccggc gaataccggt acagcatggg gtttcagtgc gagactgagg gctccggcgg
     3001 cagcggtggc tccggaggca gcggaggtac cggcggcagt ggagggtctg gtggcagcgg
     3061 aatggccgaa gggagtgtcg ccagacagcc tgacctgctg acttgcgatg acgaaccaat
     3121 ccacattccc ggcgctatcc agccccacgg actgctgctg gctctggccg ctgacatgac
     3181 tatcgtggca ggctctgata acctgccaga gctgaccggg ctggctatcg gagcactgat
     3241 tggccggtcc gcagccgacg tgttcgattc tgagactcac aatagactga ccatcgccct
     3301 ggctgaacct ggagccgccg tgggagcccc aattaccgtc ggcttcacaa tgcgaaagga
     3361 cgctggcttt atcgggtcct ggcaccgcca tgatcagctg atttttctgg agctggaacc
     3421 acctcagaga gacgtggcag agccacaggc tttctttcgg agaacaaaca gcgccatccg
     3481 gcggctccag gccgctgaga ctctggaatc cgcttgcgca gccgctgcac aggaagtgag
     3541 gaagatcacc gggttcgatc gcgtcatgat ctaccggttc gcctcagact ttagcggaga
     3601 agtgatcgca gaagatcgct gtgccgaggt cgaaagtaaa ctgggcctgc attacccagc
     3661 ctcaaccgtg cctgcacagg cacgacggct gtatacaatc aacccagtcc ggatcattcc
     3721 cgacattaat tacagaccag tgcccgtcac acctgacctg aacccagtga ctgggaggcc
     3781 catcgacctg agcttcgcca ttctgcgcag cgtgtcccct gtccacctgg agtttatgag
     3841 gaatatcgga atgcatggca ccatgtctat cagtattctg agaggcgaaa ggctgtgggg
     3901 gctgatcgtg tgccaccatc ggacacctta ctatgtggac ctggacggca ggcaggcttg
     3961 tgaactggtc gctcaggtgc tggcttggca gattggcgtg atggaagagt aatctagagg
     4021 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     4081 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     4141 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     4201 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     4261 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     4321 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4381 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4441 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4501 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4561 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4621 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4681 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4741 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4801 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4861 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4921 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4981 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     5041 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     5101 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     5161 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     5221 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     5281 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     5341 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     5401 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     5461 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     5521 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5581 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5641 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5701 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5761 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5821 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5881 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5941 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     6001 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     6061 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     6121 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     6181 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     6241 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     6301 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6361 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6421 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6481 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6541 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6601 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6661 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6721 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6781 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6841 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6901 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6961 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7021 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7081 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     7141 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7201 cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta
     7261 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7321 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7381 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7441 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7501 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7561 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7621 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7681 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7741 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7801 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7861 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7921 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7981 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8041 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8101 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8161 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8221 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8281 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8341 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8401 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
