LOCUS       Exported                7823 bp ds-DNA     circular SYN 12-MAY-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 7823)
  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 7823)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7823
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     CDS             941..1390
                     /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(3417..3434)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3423..3647
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3693..4121
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3780..3799)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3990..4011
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4130..4150)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4135..4464
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4315..4450
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4377..4396
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4531..5325
                     /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(4585..4604)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5195..5214
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5499..5620
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5536..5555)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5590..5609
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5669..5685)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5669..5685)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5682..5704)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5693..5709
                     /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(5717..5747)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5762..5783
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5900..5917)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6071..6656)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6151..6170)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6827..7687)
                     /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     7450..7469
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7688..7792)
                     /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 cacactcttt acgcccctgg
      961 aggatacgac attatgggat atttgcggca gattaggaac cgcccaaacc ctcaggtcga
     1021 actggggcct gtggacacgt catgtgccct ggtcctgtgc gatctgaagc aaaaggacac
     1081 tccggtggtc tacgcctcgg aagccttctt gtatatgacc ggatacagca atgcagaggt
     1141 gctcggcagg aactgcagat tcctgcagtc ccccgacggg atggtgaaac caaagtcgac
     1201 tcgcaaatat gtggactcga acacgatcaa caccatgcgg aaggccatcg accggaacgc
     1261 cgaggtccag gtggaggtgg tcaactttaa gaagaacggc cagcggttcg tgaactttct
     1321 gaccatgatt ccggtccggg atgaaaccgg agagtacaga tactccatgg gattccagtg
     1381 cgaaaccgaa ggttccggcg gctcgggcgg atcaggggga agcggacaca cactctacgc
     1441 cccgggaggc tacgatatca tgggctacct gaggcagatc cggaacagac cgaaccccca
     1501 agtggaactc ggccccgtgg acacttcttg cgctctggtg ctgtgcgacc tgaagcagaa
     1561 ggataccccc gtggtgtacg catccgaagc gttcctctac atgaccgggt actcgaacgc
     1621 ggaggtgctg ggtcgcaact gccggttcct gcagagcccc gacggaatgg tcaagcccaa
     1681 gtccactcgg aaatacgtgg attccaacac catcaatact atgaggaagg ctatcgatag
     1741 gaacgcggaa gtccaagtgg aggtggtgaa cttcaagaag aatggccaga gattcgtcaa
     1801 tttccttacc atgatccctg tgcgcgatga gactggagag taccgatact caatgggctt
     1861 ccagtgtgaa accgagggaa gcggcggttc cggggggtcg ggaggttccg gacataccct
     1921 ctacgcgccg gggggttatg acatcatggg ttacctcaga cagatcagaa accggccgaa
     1981 cccacaagtg gagctgggac ccgtcgacac ctcctgcgcc ctcgtgctgt gtgaccttaa
     2041 gcagaaggac acccctgtgg tgtacgcctc cgaagcattc ctgtacatga ccgggtactc
     2101 gaacgccgaa gtgctgggac ggaactgccg cttcctgcaa tccccggatg gaatggtgaa
     2161 gcctaagtca acccgcaaat acgtggactc caacactatc aacaccatgc gcaaggccat
     2221 tgaccgcaat gctgaggtgc aagtggaagt ggtgaacttc aagaagaacg gacagcgctt
     2281 cgtcaacttc ctgactatga ttcccgtgcg ggacgaaacc ggcgaatacc ggtacagcat
     2341 ggggtttcag tgcgagactg agggctccgg cggcagcggt ggctccggag gcagcggagg
     2401 taccggcggc agtggagggt ctggtggcag cggaatggcc gaagggagtg tcgccagaca
     2461 gcctgacctg ctgacttgcg atgacgaacc aatccacatt cccggcgcta tccagcccca
     2521 cggactgctg ctggctctgg ccgctgacat gactatcgtg gcaggctctg ataacctgcc
     2581 agagctgacc gggctggcta tcggagcact gattggccgg tccgcagccg acgtgttcga
     2641 ttctgagact cacaatagac tgaccatcgc cctggctgaa cctggagccg ccgtgggagc
     2701 cccaattacc gtcggcttca caatgcgaaa ggacgctggc tttatcgggt cctggcaccg
     2761 ccatgatcag ctgatttttc tggagctgga accacctcag agagacgtgg cagagccaca
     2821 ggctttcttt cggagaacaa acagcgccat ccggcggctc caggccgctg agactctgga
     2881 atccgcttgc gcagccgctg cacaggaagt gaggaagatc accgggttcg atcgcgtcat
     2941 gatctaccgg ttcgcctcag actttagcgg agaagtgatc gcagaagatc gctgtgccga
     3001 ggtcgaaagt aaactgggcc tgcattaccc agcctcaacc gtgcctgcac aggcacgacg
     3061 gctgtataca atcaacccag tccggatcat tcccgacatt aattacagac cagtgcccgt
     3121 cacacctgac ctgaacccag tgactgggag gcccatcgac ctgagcttcg ccattctgcg
     3181 cagcgtgtcc cctgtccacc tggagtttat gaggaatatc ggaatgcatg gcaccatgtc
     3241 tatcagtatt ctgagaggcg aaaggctgtg ggggctgatc gtgtgccacc atcggacacc
     3301 ttactatgtg gacctggacg gcaggcaggc ttgtgaactg gtcgctcagg tgctggcttg
     3361 gcagattggc gtgatggaag agtaatctag agggcccgtt taaacccgct gatcagcctc
     3421 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3481 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3541 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3601 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     3661 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3721 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3781 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3841 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3901 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3961 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4021 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4081 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     4141 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4201 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4261 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4321 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4381 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4441 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4501 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4561 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4621 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4681 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4741 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4801 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4861 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4921 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4981 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5041 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5101 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5161 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5221 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5281 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5341 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5401 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5461 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5521 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5581 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5641 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5701 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5761 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5821 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5881 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5941 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6001 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6061 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6121 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6181 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6241 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6301 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6361 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6421 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6481 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6541 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6601 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6661 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6721 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6781 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6841 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6901 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6961 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7021 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7081 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7141 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7201 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7261 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7321 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7381 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7441 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7501 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7561 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7621 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7681 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7741 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7801 cccgaaaagt gccacctgac gtc
//
