LOCUS Exported 7217 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses Opto-hRET (light-activated receptor tyrosine kinase) . ACCESSION . VERSION . KEYWORDS Opto-hRET_317 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7217) AUTHORS Grusch M, Schelch K, Riedler R, Reichhart E, Differ C, Berger W, Ingles-Prieto A, Janovjak H TITLE Spatio-temporally precise activation of engineered receptor tyrosine kinases by light. JOURNAL EMBO J. 2014 Jul 1. pii: e201387695. PUBMED 24986882 REFERENCE 2 (bases 1 to 7217) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7217 /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 935..944 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 941..982 /codon_start=1 /product="N-myristoylation signal from Src kinase (Pellman et al., 1985; Kaplan et al., 1988)" /label=myr /translation="MGSSKSKPKDPSQR" CDS 2708..2734 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(2811..2828) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2817..3041 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3087..3515 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3174..3193) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3384..3405 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3524..3544) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3529..3858 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3709..3844 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3771..3790 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3925..4719 /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(3979..3998) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4589..4608 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4893..5014 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4930..4949) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4984..5003 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5063..5079) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5063..5079) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5076..5098) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5087..5103 /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(5111..5141) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5156..5177 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5294..5311) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5465..6050) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5545..5564) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6221..7081) /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 6844..6863 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7082..7186) /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 ggagacacaa gctggctagc 901 gtttaaacgg gccctctaga ctcgagcggc cgcggccacc atggggagta gcaagagcaa 961 gcctaaggac cccagccagc gcctcgacgt caccggtcac tgctaccaca agtttgccca 1021 caagccaccc atctcctcag ctgagatgac cttccggagg cccgcccagg ccttcccggt 1081 cagctactcc tcttccagtg cccgccggcc ctcgctggac tccatggaga accaggtctc 1141 cgtggatgcc ttcaagatcc tggaggatcc aaagtgggaa ttccctcgga agaacttggt 1201 tcttggaaaa actctaggag aaggcgaatt tggaaaagtg gtcaaggcaa cggccttcca 1261 tctgaaaggc agagcagggt acaccacggt ggccgtgaag atgctgaaag agaacgcctc 1321 cccgagtgag cttcgagacc tgctgtcaga gttcaacgtc ctgaagcagg tcaaccaccc 1381 acatgtcatc aaattgtatg gggcctgcag ccaggatggc ccgctcctcc tcatcgtgga 1441 gtacgccaaa tacggctccc tgcggggctt cctccgcgag agccgcaaag tggggcctgg 1501 ctacctgggc agtggaggca gccgcaactc cagctccctg gaccacccgg atgagcgggc 1561 cctcaccatg ggcgacctca tctcatttgc ctggcagatc tcacagggga tgcagtatct 1621 ggccgagatg aagctcgttc atcgggactt ggcagccaga aacatcctgg tagctgaggg 1681 gcggaagatg aagatttcgg atttcggctt gtcccgagat gtttatgaag aggattcgta 1741 cgtgaagagg agccagggtc ggattccagt taaatggatg gcaattgaat ccctttttga 1801 tcatatctac accacgcaaa gtgatgtatg gtcttttggt gtcctgctgt gggagatcgt 1861 gaccctaggg ggaaacccct atcctgggat tcctcctgag cggctcttca accttctgaa 1921 gaccggccac cggatggaga ggccagacaa ctgcagcgag gagatgtacc gcctgatgct 1981 gcaatgctgg aagcaggagc cggacaaaag gccggtgttt gcggacatca gcaaagacct 2041 ggagaagatg atggttaaga ggagagacta cttggacctt gcggcgtcca ctccatctga 2101 ctccctgatt tatgacgacg gcctctcaga ggaggagaca ccgctggtgg actgtaataa 2161 tgcccccctc cctcgagccc tcccttccac atggattgaa aacaaactct atggtagaat 2221 ttcccatgca tttactagat tcaccggtgg acctgactac agtctcgtga aggctctgca 2281 aatggcacaa cagaattttg tcattacaga cgcctccctc ccagacaacc ctatcgtcta 2341 cgccagtaga gggtttctga cactgacagg ctattctctc gaccagatcc tgggcaggaa 2401 ctgcaggttt ctgcaagggc cagaaacaga cccaagagct gtggataaga tcaggaatgc 2461 catcaccaaa ggcgttgata ccagtgtctg tctgctgaat tatagacagg atggcacaac 2521 cttctggaat ctcttcttcg tggctggact cagagattct aagggcaata ttgtcaacta 2581 cgtcggagtg cagtcaaagg tgagcgaaga ttatgccaag ctgctggtca acgagcagaa 2641 cattgagtac aaaggtgtgc gcaccagtaa catgctgcgc agaaagcccg gtggatccgg 2701 agtcgactat ccgtacgacg taccagacta cgcactcgac taagaattcc accacactgg 2761 actagtggat ccgagctcgg taccaagctt aagtttaaac cgctgatcag cctcgactgt 2821 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga 2881 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag 2941 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga 3001 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac 3061 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg 3121 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 3181 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 3241 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 3301 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 3361 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 3421 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa 3481 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta 3541 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 3601 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 3661 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 3721 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 3781 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga 3841 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa 3901 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg 3961 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 4021 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 4081 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg 4141 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 4201 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 4261 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 4321 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 4381 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 4441 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc 4501 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 4561 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 4621 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 4681 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa 4741 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct 4801 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 4861 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt 4921 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 4981 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta 5041 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 5101 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 5161 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 5221 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 5281 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 5341 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 5401 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 5461 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 5521 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 5581 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 5641 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 5701 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 5761 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 5821 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 5881 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 5941 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 6001 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 6061 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 6121 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 6181 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 6241 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 6301 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 6361 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 6421 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 6481 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 6541 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 6601 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 6661 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 6721 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 6781 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 6841 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 6901 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 6961 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 7021 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 7081 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 7141 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 7201 aagtgccacc tgacgtc //