LOCUS Exported 6635 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 6635) 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 6635) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6635 /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 1427..2143 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2229..2246) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2235..2459 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2505..2933 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2592..2611) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2802..2823 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2942..2962) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2947..3276 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3127..3262 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3189..3208 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3343..4137 /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(3397..3416) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4007..4026 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4311..4432 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4348..4367) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4402..4421 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4481..4497) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4481..4497) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4494..4516) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4505..4521 /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(4529..4559) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4574..4595 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4712..4729) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4883..5468) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4963..4982) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5639..6499) /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 6262..6281 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6500..6604) /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 aagcttatgg gacacactct ttacgcccct ggaggatacg acattatggg 961 atatttggat cagattgcga accgcccaaa ccctcaggtc gaactggggc ctgtggacct 1021 gtcatgtgcc ctgatcctgt gcgatctgaa gcaaaaggac actccgatcg tctacgcctc 1081 ggaagccttc ttggagatga ccggatacaa cagacatgag gtgctcggca ggaactgcag 1141 attcctgcag tcccccgacg ggatggtgaa accaaagtcg actcgcaaat atgtggactc 1201 gaacacgatc ttcaccatca agaaggccat cgaccggaac gccgaggtcc aggtggaggt 1261 ggtcaacttt aagaagaacg gccagcggtt cgtgaacttt ctgaccatca ttccggtccg 1321 ggatgaaacc ggagagtaca gatactccat cggattccag tgcgaaaccg aaaccggtgg 1381 cggctccgga ggcagcggcg gttctggagg ttccggtggc ggctccatgg tgtcgaaggg 1441 agaggagctt ttcaccggcg tggtgccgat tcttgtggaa ctggacggcg acgtgaacgg 1501 acataagttc agcgtgtcag gagaaggaga aggggacgcg acttacggaa agctgaccct 1561 gaagttcatc tgtaccaccg ggaagctccc ggtgccctgg ccgaccctcg tgactactct 1621 gacctacgga gtgcagtgct tctcccgcta cccggaccat atgaagcagc acgatttctt 1681 caagagcgct atgcccgagg gttacgtgca agaacggacc atcttcttca aggacgacgg 1741 aaactacaag accagagccg aagtgaaatt cgaaggagac accctcgtca atcgcatcga 1801 gctgaaggga atcgacttta aggaagatgg gaatatcctg ggtcacaagc tggagtacaa 1861 ctacaactcc cacaatgtgt acatcatggc ggataagcaa aagaacggca ttaaggtcaa 1921 cttcaagatc cggcataaca tagaagatgg ctccgtgcaa ctggcggacc actaccagca 1981 gaacaccccc attggggacg gtccggtgct gctccctgat aaccactacc tgtcaaccca 2041 aagcgcactg tccaaggacc ccaacgagaa gagggaccac atggtgctgc tggagttcgt 2101 cactgccgct ggcatcactc tcggaatgga cgagctgtac aagggggata tcaagaaaaa 2161 gaagaagaag tccaagacaa aatgcgtgat tatgtagtct agagggcccg tttaaacccg 2221 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt 2281 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat 2341 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag 2401 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc 2461 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg 2521 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 2581 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 2641 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 2701 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac 2761 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 2821 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat 2881 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg 2941 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg 3001 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca 3061 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact 3121 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 3181 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag 3241 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc 3301 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga 3361 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa 3421 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt 3481 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg 3541 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa 3601 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac 3661 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt 3721 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact 3781 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg 3841 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg 3901 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc 3961 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt 4021 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc 4081 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg 4141 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 4201 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 4261 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta 4321 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 4381 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 4441 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt 4501 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 4561 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 4621 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 4681 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 4741 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 4801 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 4861 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 4921 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 4981 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 5041 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 5101 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 5161 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 5221 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 5281 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 5341 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 5401 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 5461 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 5521 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 5581 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 5641 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 5701 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 5761 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 5821 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 5881 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 5941 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 6001 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 6061 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 6121 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 6181 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 6241 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 6301 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 6361 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 6421 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 6481 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 6541 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 6601 cgcgcacatt tccccgaaaa gtgccacctg acgtc //