LOCUS Exported 6660 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses Opto-casp9_VfAU1-LOV_casp9 (light-activated caspase9). ACCESSION . VERSION . KEYWORDS Opto-casp9_VfAU1-LOV_casp9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6660) 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 6660) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6660 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 170..549 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 550..753 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 704..724 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 798..817 /label=T7 /note="T7 promoter, forward primer" promoter 798..816 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS complement(1640..1651) /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /translation="WELQ" primer_bind complement(2186..2203) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2192..2416 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2462..2890 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2549..2568) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2759..2780 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2899..2919) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2904..3233 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3084..3219 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3146..3165 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3300..4094 /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(3354..3373) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3964..3983 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4268..4389 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4305..4324) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4359..4378 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4438..4454) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4438..4454) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4451..4473) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4462..4478 /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(4486..4516) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4531..4552 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4669..4686) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4840..5428) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4920..4939) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5599..6459) /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 6222..6241 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6460..6564) /gene="bla" /label=AmpR promoter primer_bind complement(6639..6658) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc 61 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg 121 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 181 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 241 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 301 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 361 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 421 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 481 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 541 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 601 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 661 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 721 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct 781 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctga ccatggccac 841 cggtcctgac tacagtctcg tgaaggctct gcaaatggca caacagaatt ttgtcattac 901 agacgcctcc ctcccagaca accctatcgt ctacgccagt agagggtttc tgacactgac 961 aggctattct ctcgaccaga tcctgggcag gaactgcagg tttctgcaag ggccagaaac 1021 agacccaaga gctgtggata agatcaggaa tgccatcacc aaaggcgttg ataccagtgt 1081 ctgtctgctg aattatagac aggatggcac aaccttctgg aatctcttct tcgtggctgg 1141 actcagagat tctaagggca atattgtcaa ctacgtcgga gtgcagtcaa aggtgagcga 1201 agattatgcc aagctgctgg tcaacgagca gaacattgag tacaaaggtg tgcgcaccag 1261 taacatgctg cgcagaaagc ccggtgaatt ctcaggctct ggatcccccg ggggatttgg 1321 tgatgtcggt gctcttgaga gtttgagggg aaatgcagat ttggcttaca tcctgagcat 1381 ggagccctgt ggccactgcc tcattatcaa caatgtgaac ttctgccgtg agtccgggct 1441 ccgcacccgc actggctcca acatcgactg tgagaagttg cggcgtcgct tctcctcgct 1501 gcatttcatg gtggaggtga agggcgacct gactgccaag aaaatggtgc tggctttgct 1561 ggagctggcg cggcaggacc acggtgctct ggactgctgc gtggtggtca ttctctctca 1621 cggctgtcag gccagccacc tgcagttccc aggggctgtc tacggcacag atggatgccc 1681 tgtgtcggtc gagaagattg tgaacatctt caatgggacc agctgcccca gcctgggagg 1741 gaagcccaag ctctttttca tccaggcctg tggtggggag cagaaagacc atgggtttga 1801 ggtggcctcc acttcccctg aagacgagtc ccctggcagt aaccccgagc cagatgccac 1861 cccgttccag gaaggtttga ggaccttcga ccagctggac gccatatcta gtttgcccac 1921 acccagtgac atctttgtgt cctactctac tttcccaggt tttgtttcct ggagggaccc 1981 caagagtggc tcctggtacg ttgagaccct ggacgacatc tttgagcagt gggctcactc 2041 tgaagacctg cagtccctcc tgcttagggt cgctaatgct gtttcggtga aagggattta 2101 taaacagatg cctggttgct ttaatttcct ccggaaaaaa cttttcttta aaacatcacc 2161 cgggtaagtt taaaccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt 2221 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc 2281 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt 2341 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat 2401 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc 2461 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 2521 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 2581 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt 2641 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg 2701 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt 2761 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta 2821 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt 2881 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc 2941 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt 3001 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 3061 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc 3121 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg 3181 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctct 3241 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca 3301 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg 3361 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag 3421 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc 3481 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc 3541 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg 3601 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc 3661 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca 3721 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag 3781 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg 3841 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg 3901 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca 3961 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc 4021 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg 4081 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct 4141 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt 4201 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc 4261 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 4321 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 4381 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc 4441 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 4501 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 4561 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 4621 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 4681 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 4741 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 4801 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 4861 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 4921 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 4981 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 5041 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 5101 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 5161 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 5221 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 5281 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 5341 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 5401 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 5461 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 5521 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 5581 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 5641 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 5701 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 5761 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 5821 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 5881 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 5941 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 6001 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 6061 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 6121 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 6181 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 6241 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 6301 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 6361 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 6421 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 6481 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 6541 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg 6601 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca //