LOCUS Exported 7818 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses Opto-casp9_AtCRY2-PHR-casp9 (light-activated caspase9). ACCESSION . VERSION . KEYWORDS Opto-casp9_AtCRY2-PHR-casp9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7818) AUTHORS Tichy AM, Gerrard EJ, Legrand JMD, Hobbs RM, Janovjak H TITLE Engineering Strategy and Vector Library for the Rapid Generation of Modular Light-Controlled Protein-Protein Interactions. JOURNAL J Mol Biol. 2019 May 29. pii: S0022-2836(19)30317-1. doi: 10.1016/j.jmb.2019.05.033. PUBMED 31150735 REFERENCE 2 (bases 1 to 7818) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7818 /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 2413..2439 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2440..2496 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 2497..2526 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS complement(2866..2877) /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /translation="WELQ" primer_bind complement(3412..3429) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3418..3642 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3688..4116 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3775..3794) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3985..4006 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4125..4145) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4130..4459 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4310..4445 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4372..4391 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4526..5320 /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(4580..4599) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5190..5209 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5494..5615 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5531..5550) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5585..5604 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5664..5680) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5664..5680) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5677..5699) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5688..5704 /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(5712..5742) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5757..5778 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5895..5912) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6066..6651) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6146..6165) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6822..7682) /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 7445..7464 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7683..7787) /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 gctgaccatg 901 gccaccggta agatggacaa gaaaaccata gtctggtttc gccgcgattt gaggatagag 961 gataaccctg cgttggccgc agcggcccac gaaggcagcg tgttccccgt gttcatatgg 1021 tgcccagagg aggaaggcca gttctaccca ggtcgcgcta gtcgctggtg gatgaaacag 1081 tctctcgcac atctctccca atctttgaaa gctctcgggt ctgaccttac gctgattaag 1141 acccacaata ctattagtgc aatcttggac tgcatccgcg ttaccggcgc gaccaaagtg 1201 gtgtttaatc atctgtatga cccggtaagt ctggtacgcg accatacagt taaggagaag 1261 ttggttgaaa ggggaatttc agtacagagt tataatggag acctcctcta cgaaccttgg 1321 gaaatatact gtgagaaggg aaagccattt acatcattta actcatactg gaaaaagtgt 1381 cttgacatga gcatagagtc tgtcatgttg cccccgccgt ggcggcttat gcccatcacc 1441 gcggccgccg aagctatctg ggcgtgtagt attgaagaac ttggcttgga gaacgaagca 1501 gaaaaaccta gcaatgcact gttgacgcgc gcctggtccc ctgggtggag taacgcagat 1561 aaattgctta acgagttcat cgaaaagcaa cttatcgact acgctaagaa tagcaaaaaa 1621 gtggtgggca attcaacctc cctcctgtct ccgtacttgc atttcggcga gattagcgtg 1681 cggcacgtct tccaatgcgc ccgaatgaaa caaataatct gggcgcggga taaaaacagt 1741 gagggtgaag aaagcgcaga cttgttcctc cgaggaatcg gtctgcgaga atatagtcgc 1801 tacatttgtt tcaattttcc ctttacgcat gagcagagcc tccttagtca cttgcgattc 1861 tttccttggg acgcagatgt tgacaaattt aaagcatggc ggcaaggtag aacaggctac 1921 ccattggtag atgcgggtat gcgagaactc tgggccacgg ggtggatgca taaccgaatc 1981 agggtaatag taagtagttt cgcagttaag tttcttttgc ttccatggaa gtgggggatg 2041 aagtatttct gggacacttt gctcgatgcg gatcttgaat gcgatatatt gggttggcaa 2101 tatatttccg ggtcaatccc tgacggccat gagctggaca gattggacaa tcctgcgctc 2161 cagggtgcga aatacgatcc cgaaggtgaa tacattagac aatggcttcc agaacttgcc 2221 aggttgccca cggaatggat tcaccaccca tgggacgccc ctcttacggt tttgaaggcg 2281 agtggtgtgg agctcggtac caattacgca aaaccgattg ttgacattga taccgcgcgg 2341 gagttgctgg ctaaagccat ttcacgaacc cggaaagccc aaatcatgat tggtgctgca 2401 cccggtgaat tctacccata cgatgttcca gattacgctg cgaccaactt tagcctgctg 2461 aaacaggcgg gcgatgtgga agaaaacccg ggcccggaac aaaaactcat ctcagaagag 2521 gatctgggat cccccggggg atttggtgat gtcggtgctc ttgagagttt gaggggaaat 2581 gcagatttgg cttacatcct gagcatggag ccctgtggcc actgcctcat tatcaacaat 2641 gtgaacttct gccgtgagtc cgggctccgc acccgcactg gctccaacat cgactgtgag 2701 aagttgcggc gtcgcttctc ctcgctgcat ttcatggtgg aggtgaaggg cgacctgact 2761 gccaagaaaa tggtgctggc tttgctggag ctggcgcggc aggaccacgg tgctctggac 2821 tgctgcgtgg tggtcattct ctctcacggc tgtcaggcca gccacctgca gttcccaggg 2881 gctgtctacg gcacagatgg atgccctgtg tcggtcgaga agattgtgaa catcttcaat 2941 gggaccagct gccccagcct gggagggaag cccaagctct ttttcatcca ggcctgtggt 3001 ggggagcaga aagaccatgg gtttgaggtg gcctccactt cccctgaaga cgagtcccct 3061 ggcagtaacc ccgagccaga tgccaccccg ttccaggaag gtttgaggac cttcgaccag 3121 ctggacgcca tatctagttt gcccacaccc agtgacatct ttgtgtccta ctctactttc 3181 ccaggttttg tttcctggag ggaccccaag agtggctcct ggtacgttga gaccctggac 3241 gacatctttg agcagtgggc tcactctgaa gacctgcagt ccctcctgct tagggtcgct 3301 aatgctgttt cggtgaaagg gatttataaa cagatgcctg gttgctttaa tttcctccgg 3361 aaaaaacttt tctttaaaac atcacccggg taagtttaaa ccgctgatca gcctcgactg 3421 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg 3481 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga 3541 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg 3601 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa 3661 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg 3721 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt 3781 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc 3841 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg 3901 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga 3961 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc 4021 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa 4081 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt 4141 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 4201 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 4261 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct 4321 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc 4381 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg 4441 aggcctaggc ttttgcaaaa agctctcggg agcttgtata tccattttcg gatctgatca 4501 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc 4561 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc 4621 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga 4681 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac 4741 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct 4801 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa 4861 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc 4921 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct 4981 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc 5041 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg 5101 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct 5161 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct 5221 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca 5281 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa 5341 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc 5401 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc 5461 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt 5521 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 5581 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct 5641 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 5701 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 5761 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 5821 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 5881 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 5941 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 6001 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 6061 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 6121 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 6181 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 6241 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 6301 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 6361 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 6421 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 6481 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca 6541 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 6601 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 6661 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 6721 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 6781 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 6841 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 6901 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 6961 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 7021 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 7081 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 7141 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 7201 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 7261 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 7321 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 7381 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 7441 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 7501 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 7561 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 7621 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 7681 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 7741 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 7801 aaagtgccac ctgacgtc //