LOCUS Exported 8453 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Blue light dependent photoswitching system. ACCESSION . VERSION . KEYWORDS pcDNA3.1_iSH2-pMag(3x)-iRFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8453) 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 8453) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8453 /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" regulatory 917..926 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" 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=pMag /note="rapidly heterodimerizes with nMagHigh1 upon blue light irradiation to form a complex with slow switch-off kinetics" /translation="HTLYAPGGYDIMGYLRQIRNRPNPQVELGPVDTSCALILCDLKQK DTPIVYASEAFLYMTGYSNAEVLGRNCRFLQSPDGMVKPKSTRKYVDSNTINTMRKAID 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(6344..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..7286) /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(7457..8317) /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 8080..8099 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8318..8422) /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 gtgccctgat 1681 cctgtgcgat ctgaagcaaa aggacactcc gatcgtctac 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 tctgatcctg tgcgacctga agcagaagga tacccccatc 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 atcctgtgtg accttaagca gaaggacacc cctatcgtgt 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 gcggtggttt ttttgtttgc aagcagcaga 7261 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 7321 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 7381 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 7441 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 7501 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg 7561 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag 7621 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt 7681 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag 7741 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt 7801 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca 7861 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg 7921 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat 7981 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta 8041 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca 8101 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct 8161 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat 8221 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa 8281 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt 8341 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa 8401 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc //