LOCUS Exported 8368 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses Opto-hEGFR (light-activated receptor tyrosine kinase). ACCESSION . VERSION . KEYWORDS Opto-hEGFR_308 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8368) 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 8368) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8368 /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 3859..3885 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(3962..3979) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3968..4192 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4238..4666 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4325..4344) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4535..4556 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4675..4695) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4680..5009 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4860..4995 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4922..4941 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5076..5870 /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(5130..5149) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5740..5759 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6044..6165 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6081..6100) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6135..6154 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6214..6230) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6214..6230) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6227..6249) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6238..6254 /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(6262..6292) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6307..6328 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6445..6462) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6616..7201) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6696..6715) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7372..8232) /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 7995..8014 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8233..8337) /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 cgcaccatgg gggcaggcgc aactggtaga 961 gcaatggacg gacctagact cctgctgctc ctcctgctcg gagtgtccct gggtggagca 1021 aaagaagcat gcccaactgg actctacacc cattctggag agtgctgcaa ggcctgcaat 1081 ctgggtgaag gagtcgctca gccatgtgga gcaaaccaga ccgtctgtga gccttgcctg 1141 gattctgtga cattctccga cgtggtgtct gccaccgagc cttgcaagcc atgcaccgag 1201 tgcgttggcc tgcagtcaat gtctgctccc tgcgtggagg ctgacgacgc agtgtgtaga 1261 tgcgcatatg gctactacca ggatgaaact actgggaggt gcgaggcatg tagggtgtgt 1321 gaggcaggca gtggtctggt tttcagttgc caggacaagc agaacaccgt ctgcgaggag 1381 tgtccagacg ggacatacag cgacgaagcc aaccatgtcg atccctgcct gccatgtacc 1441 gtttgcgaag ataccgaacg ccagctgaga gaatgcacta gatgggctga cgccgaatgt 1501 gaggagatcc caggacggtg gattactcgc tctactcctc cagaagggtc agatagcacc 1561 gctcccagca cccaagaacc cgaagctcct cccgagcagg atctgattgc ctccacagtt 1621 gctggagtcg tgactaccgt gatgggttcc tcacaacccg tcgtgaccag agggacaacc 1681 gacaacctga ttccagtgta ttgctctatt ctggctgccg tcgtggtggg actggttgca 1741 tacatagcct tcaagagggg gcgcgcccga aggcgccaca tcgttcggaa gcgcacgctg 1801 cggaggctgc tgcaggagag ggagcttgtg gagcctctta cacccagtgg agaagctccc 1861 aaccaagctc tcttgaggat cttgaaggaa actgaattca aaaagatcaa agtgctgggc 1921 tccggtgcgt tcggcacggt gtataaggga ctctggatcc cagaaggtga gaaagttaaa 1981 attcccgtcg ctatcaagga attaagagaa gcaacatctc cgaaagccaa caaggaaatc 2041 ctcgatgaag cctacgtgat ggccagcgtg gacaaccccc acgtgtgccg cctgctgggc 2101 atctgcctca cctccaccgt gcaactcatc acgcagctca tgcccttcgg ctgcctcctg 2161 gactatgtcc gggaacacaa agacaatatt ggctcccagt acctgctcaa ctggtgtgtg 2221 cagatcgcaa agggcatgaa ctacttggag gaccgtcgct tggtgcaccg cgacctggca 2281 gccaggaacg tactggtgaa aacaccgcag catgtcaaga tcacagattt tgggctggcc 2341 aaactgctgg gtgcggaaga gaaagaatac catgcagaag gaggcaaagt gcctatcaag 2401 tggatggcat tggaatcaat tttacacaga atctataccc accagagtga tgtctggagc 2461 tacggggtga ccgtttggga gttgatgacc tttggatcca agccatatga cggaatccct 2521 gccagcgaga tctcctccat cctggagaaa ggagaacgcc tccctcagcc acccatatgt 2581 accatcgatg tctacatgat catggtcaag tgctggatga tagacgcaga tagtcgccca 2641 aagttccgtg agttgatcat cgaattctcc aaaatggccc gagaccccca gcgctacctt 2701 gtcattcagg gggatgaaag aatgcatttg ccaagtccta cagactccaa cttctaccgt 2761 gccctgatgg atgaagaaga catggacgac gtggtggatg ccgacgagta cctcatccca 2821 cagcagggct tcttcagcag cccctccacg tcacggactc ccctcctgag ctctctgagt 2881 gcaaccagca acaattccac cgtggcttgc attgatagaa atgggctgca aagctgtccc 2941 atcaaggaag acagcttctt gcagcgatac agctcagacc ccacaggcgc cttgactgag 3001 gacagcatag acgacacctt cctcccagtg cctgaataca taaaccagtc cgttcccaaa 3061 aggcccgctg gctctgtgca gaatcctgtc tatcacaatc agcctctgaa ccccgcgccc 3121 agcagagacc cacactacca ggacccccac agcactgcag tgggcaaccc cgagtatctc 3181 aacactgtcc agcccacctg tgtcaacagc acattcgaca gccctgccca ctgggcccag 3241 aaaggcagcc accaaattag cctggacaac cctgactacc agcaggactt ctttcccaag 3301 gaagccaagc caaatggcat ctttaagggc tccacagctg aaaatgcaga atacctaagg 3361 gtcgcgccac aaagcagtga atttattgga gcatccggtg gacctgacta cagtctcgtg 3421 aaggctctgc aaatggcaca acagaatttt gtcattacag acgcctccct cccagacaac 3481 cctatcgtct acgccagtag agggtttctg acactgacag gctattctct cgaccagatc 3541 ctgggcagga actgcaggtt tctgcaaggg ccagaaacag acccaagagc tgtggataag 3601 atcaggaatg ccatcaccaa aggcgttgat accagtgtct gtctgctgaa ttatagacag 3661 gatggcacaa ccttctggaa tctcttcttc gtggctggac tcagagattc taagggcaat 3721 attgtcaact acgtcggagt gcagtcaaag gtgagcgaag attatgccaa gctgctggtc 3781 aacgagcaga acattgagta caaaggtgtg cgcaccagta acatgctgcg cagaaagccc 3841 ggtggatccg gagtcgacta tccgtacgac gtaccagact acgcactcga ctaagaattc 3901 caccacactg gactagtgga tccgagctcg gtaccaagct taagtttaaa ccgctgatca 3961 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc 4021 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg 4081 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg 4141 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag 4201 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta 4261 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg 4321 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa 4381 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc 4441 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt 4501 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca 4561 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc 4621 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg 4681 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 4741 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa 4801 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca 4861 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt 4921 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag 4981 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg 5041 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg 5101 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa 5161 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg 5221 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt 5281 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa 5341 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc 5401 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg 5461 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg 5521 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg 5581 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg 5641 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact 5701 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg 5761 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc 5821 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct 5881 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac 5941 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat 6001 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc 6061 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 6121 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc 6181 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg 6241 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg 6301 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc 6361 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt 6421 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct 6481 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga 6541 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc 6601 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg 6661 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg 6721 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt 6781 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt 6841 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg 6901 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact 6961 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt 7021 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct 7081 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac 7141 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 7201 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 7261 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 7321 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 7381 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 7441 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 7501 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 7561 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 7621 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 7681 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 7741 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 7801 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 7861 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 7921 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 7981 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 8041 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 8101 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 8161 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 8221 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 8281 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 8341 atttccccga aaagtgccac ctgacgtc //