LOCUS Exported 6638 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Blue-light dependent heterodimerization system. ACCESSION . VERSION . KEYWORDS eMagAF-EGFP-PM SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6638) AUTHORS Benedetti L, Marvin JS, Falahati H, Guillen-Samander A, Looger LL, De Camilli P TITLE Optimized Vivid-derived Magnets photodimerizers for subcellular optogenetics in mammalian cells. JOURNAL Elife. 2020 Nov 11;9. pii: 63230. doi: 10.7554/eLife.63230. PUBMED 33174843 REFERENCE 2 (bases 1 to 6638) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6638 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(49..68) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 240..619 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 620..823 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 774..794 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 868..887 /label=T7 /note="T7 promoter, forward primer" promoter 868..886 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1432..2148 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2234..2251) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2240..2464 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2510..2938 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2597..2616) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2807..2828 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2947..2967) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2952..3281 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3132..3267 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3194..3213 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3348..4142 /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(3402..3421) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4012..4031 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4316..4437 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4353..4372) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4407..4426 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4486..4502) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4486..4502) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4499..4521) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4510..4526 /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(4534..4564) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4579..4600 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4717..4734) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4888..5476) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4968..4987) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5647..6507) /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 6270..6289 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6508..6612) /gene="bla" /label=AmpR promoter ORIGIN 1 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc 61 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta 121 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa 181 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg 241 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc 301 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa 361 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac 421 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca 481 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg 541 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt 601 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg 661 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg 721 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat 781 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga 841 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg 901 ctagcgttta aacttaagct tatgggacac actctttacg cccctggagg atacgacatt 961 atgggatatt tggatcagat tgcgaaccgc ccaaaccctc aggtcgaact ggggcctgtg 1021 gacctgtcat gtgccctgat cctgtgcgat ctgaagcaaa aggacactcc gatcgtctac 1081 gcctcggaag ccttcttgga gatgaccgga tacaacagac atgaggtgct cggcaggaac 1141 tgcagattcc tgcagtcccc cgacgggatg gtgaaaccaa agtcgactcg caaatatgtg 1201 gactcgaaca cgatcttcac catcaagaag gccatcgacc ggaacgccga ggtccaggtg 1261 gaggtggtca actttaagaa gaacggccag cggttcgtga actttctgac catcattccg 1321 gtccgggatg aaaccggaga gtacagatac tccatcggat tccagtgcga aaccgaaacc 1381 ggtggcggct ccggaggcag cggcggttct ggaggttccg gtggcggctc catggtgtcg 1441 aagggagagg agcttttcac cggcgtggtg ccgattcttg tggaactgga cggcgacgtg 1501 aacggacata agttcagcgt gtcaggagaa ggagaagggg acgcgactta cggaaagctg 1561 accctgaagt tcatctgtac caccgggaag ctcccggtgc cctggccgac cctcgtgact 1621 actctgacct acggagtgca gtgcttctcc cgctacccgg accatatgaa gcagcacgat 1681 ttcttcaaga gcgctatgcc cgagggttac gtgcaagaac ggaccatctt cttcaaggac 1741 gacggaaact acaagaccag agccgaagtg aaattcgaag gagacaccct cgtcaatcgc 1801 atcgagctga agggaatcga ctttaaggaa gatgggaata tcctgggtca caagctggag 1861 tacaactaca actcccacaa tgtgtacatc atggcggata agcaaaagaa cggcattaag 1921 gtcaacttca agatccggca taacatagaa gatggctccg tgcaactggc ggaccactac 1981 cagcagaaca cccccattgg ggacggtccg gtgctgctcc ctgataacca ctacctgtca 2041 acccaaagcg cactgtccaa ggaccccaac gagaagaggg accacatggt gctgctggag 2101 ttcgtcactg ccgctggcat cactctcgga atggacgagc tgtacaaggg ggatatcaag 2161 aaaaagaaga agaagtccaa gacaaaatgc gtgattatgt agtctagagg gcccgtttaa 2221 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc 2281 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag 2341 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag 2401 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct 2461 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt 2521 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc 2581 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc 2641 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg 2701 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga 2761 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc 2821 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg 2881 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa 2941 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa 3001 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc 3061 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc 3121 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct 3181 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga 3241 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta 3301 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag 3361 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg 3421 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc 3481 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag 3541 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca 3601 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat 3661 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata 3721 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac 3781 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc 3841 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg 3901 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg 3961 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta 4021 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg 4081 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct 4141 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga 4201 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc 4261 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt 4321 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 4381 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 4441 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc 4501 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 4561 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 4621 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 4681 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 4741 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 4801 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 4861 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 4921 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 4981 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 5041 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 5101 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 5161 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 5221 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 5281 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg 5341 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 5401 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 5461 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 5521 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 5581 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 5641 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 5701 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 5761 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 5821 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 5881 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 5941 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 6001 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 6061 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 6121 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 6181 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 6241 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 6301 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 6361 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 6421 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 6481 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 6541 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 6601 aggggttccg cgcacatttc cccgaaaagt gccacctg //