LOCUS       Exported                7826 bp ds-DNA     circular SYN 20-DEC-2021
DEFINITION  Blue light dependent photoswitching system.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3.1_pMagFast2(3x)-iRFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7826)
  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 7826)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 20, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7826
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(32..136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(211..230)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        402..781
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        782..985
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     936..956
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1030..1049
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1030..1048
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1108..1557
                     /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(3584..3601)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3590..3814
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3860..4288
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3947..3966)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4157..4178
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4297..4317)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4302..4631
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4482..4617
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4544..4563
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4698..5492
                     /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(4752..4771)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5362..5381
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5666..5787
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5703..5722)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5757..5776
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5836..5852)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5836..5852)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5849..5871)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5860..5876
                     /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(5884..5914)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5929..5950
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6067..6084)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6238..6826)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6318..6337)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6997..31)
                     /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     7620..7639
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
       61 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
      121 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
      181 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
      241 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
      301 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
      361 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
      421 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      481 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      541 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      601 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      661 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      721 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      781 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      841 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      901 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      961 gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg cttactggct
     1021 tatcgaaatt aatacgactc actataggga gacccaagct ggctagcgtt taaacttaag
     1081 cttaagctgg ctagcgccgc caccatgcac actctttacg cccctggagg atacgacatt
     1141 atgggatatt tgcggcagat taggaaccgc ccaaaccctc aggtcgaact ggggcctgtg
     1201 gacacgtcat gtgccctggt cctgtgcgat ctgaagcaaa aggacactcc ggtggtctac
     1261 gcctcggaag ccttcttgta tatgaccgga tacagcaatg cagaggtgct cggcaggaac
     1321 tgcagattcc tgcagtcccc cgacgggatg gtgaaaccaa agtcgactcg caaatatgtg
     1381 gactcgaaca cgatcaacac catgcggaag gccatcgacc ggaacgccga ggtccaggtg
     1441 gaggtggtca actttaagaa gaacggccag cggttcgtga actttctgac catgattccg
     1501 gtccgggatg aaaccggaga gtacagatac tccatgggat tccagtgcga aaccgaaggt
     1561 tccggcggct cgggcggatc agggggaagc ggacacacac tctacgcccc gggaggctac
     1621 gatatcatgg gctacctgag gcagatccgg aacagaccga acccccaagt ggaactcggc
     1681 cccgtggaca cttcttgcgc tctggtgctg tgcgacctga agcagaagga tacccccgtg
     1741 gtgtacgcat ccgaagcgtt cctctacatg accgggtact cgaacgcgga ggtgctgggt
     1801 cgcaactgcc ggttcctgca gagccccgac ggaatggtca agcccaagtc cactcggaaa
     1861 tacgtggatt ccaacaccat caatactatg aggaaggcta tcgataggaa cgcggaagtc
     1921 caagtggagg tggtgaactt caagaagaat ggccagagat tcgtcaattt ccttaccatg
     1981 atccctgtgc gcgatgagac tggagagtac cgatactcaa tgggcttcca gtgtgaaacc
     2041 gagggaagcg gcggttccgg ggggtcggga ggttccggac ataccctcta cgcgccgggg
     2101 ggttatgaca tcatgggtta cctcagacag atcagaaacc ggccgaaccc acaagtggag
     2161 ctgggacccg tcgacacctc ctgcgccctc gtgctgtgtg accttaagca gaaggacacc
     2221 cctgtggtgt acgcctccga agcattcctg tacatgaccg ggtactcgaa cgccgaagtg
     2281 ctgggacgga actgccgctt cctgcaatcc ccggatggaa tggtgaagcc taagtcaacc
     2341 cgcaaatacg tggactccaa cactatcaac accatgcgca aggccattga ccgcaatgct
     2401 gaggtgcaag tggaagtggt gaacttcaag aagaacggac agcgcttcgt caacttcctg
     2461 actatgattc ccgtgcggga cgaaaccggc gaataccggt acagcatggg gtttcagtgc
     2521 gagactgagg gctccggcgg cagcggtggc tccggaggca gcggaggtac cggcggcagt
     2581 ggagggtctg gtggcagcgg aatggccgaa gggagtgtcg ccagacagcc tgacctgctg
     2641 acttgcgatg acgaaccaat ccacattccc ggcgctatcc agccccacgg actgctgctg
     2701 gctctggccg ctgacatgac tatcgtggca ggctctgata acctgccaga gctgaccggg
     2761 ctggctatcg gagcactgat tggccggtcc gcagccgacg tgttcgattc tgagactcac
     2821 aatagactga ccatcgccct ggctgaacct ggagccgccg tgggagcccc aattaccgtc
     2881 ggcttcacaa tgcgaaagga cgctggcttt atcgggtcct ggcaccgcca tgatcagctg
     2941 atttttctgg agctggaacc acctcagaga gacgtggcag agccacaggc tttctttcgg
     3001 agaacaaaca gcgccatccg gcggctccag gccgctgaga ctctggaatc cgcttgcgca
     3061 gccgctgcac aggaagtgag gaagatcacc gggttcgatc gcgtcatgat ctaccggttc
     3121 gcctcagact ttagcggaga agtgatcgca gaagatcgct gtgccgaggt cgaaagtaaa
     3181 ctgggcctgc attacccagc ctcaaccgtg cctgcacagg cacgacggct gtatacaatc
     3241 aacccagtcc ggatcattcc cgacattaat tacagaccag tgcccgtcac acctgacctg
     3301 aacccagtga ctgggaggcc catcgacctg agcttcgcca ttctgcgcag cgtgtcccct
     3361 gtccacctgg agtttatgag gaatatcgga atgcatggca ccatgtctat cagtattctg
     3421 agaggcgaaa ggctgtgggg gctgatcgtg tgccaccatc ggacacctta ctatgtggac
     3481 ctggacggca ggcaggcttg tgaactggtc gctcaggtgc tggcttggca gattggcgtg
     3541 atggaagagt aatctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     3601 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3661 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3721 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3781 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3841 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3901 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3961 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4021 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4081 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4141 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4201 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4261 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     4321 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4381 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4441 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4501 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4561 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4621 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     4681 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4741 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4801 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4861 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4921 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4981 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     5041 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5101 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5161 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5221 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5281 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5341 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5401 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5461 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5521 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5581 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5641 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5701 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5761 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5821 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5881 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5941 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6001 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6061 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6121 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6181 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6241 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6301 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6361 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6421 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6481 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6541 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6601 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6661 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6721 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6781 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6841 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6901 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6961 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7021 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7081 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7141 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7201 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7261 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7321 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7381 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7441 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7501 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7561 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7621 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7681 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7741 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7801 aaggcaaaat gccgcaaaaa agggaa
//
