LOCUS Exported 8368 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses RIG-I with a mutation to motif III in mammalian cells. ACCESSION . VERSION . KEYWORDS pEF-Bos-FLAG RIG-I MIII SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8368) AUTHORS Bamming D, Horvath CM TITLE Regulation of signal transduction by enzymatically inactive antiviral RNA helicase proteins MDA5, RIG-I, and LGP2. JOURNAL J Biol Chem. 2009 Apr 10;284(15):9700-12. doi: 10.1074/jbc.M807365200. Epub 2009 Feb 11. PUBMED 19211564 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" rep_origin 1..456 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(88..107) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 298..319 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(473..492) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 592..614 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(652..670) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 738..842 /gene="bla" /label=AmpR promoter CDS 843..1703 /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 complement(1061..1080) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1874..2462 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2363..2382 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2616..2633 /label=L4440 /note="L4440 vector, forward primer" protein_bind 2750..2771 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2786..2816 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2824..2840 /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)." primer_bind 2829..2851 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2848..2864 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2848..2864 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 2896..3106 /label=SV40 promoter /note="SV40 early promoter" promoter 2918..3114 /label=SV40 promoter /note="SV40 early promoter" rep_origin 2965..3100 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3027..3046 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 3201..4379 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 3432..4370 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 4327..4347 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 4396..4405 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4405..4428 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 4432..4455 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" primer_bind complement(8140..8157) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(8140..8156) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8149..8171) /label=M13/pUC Forward /note="In lacZ gene" ORIGIN 1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 61 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 181 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc 241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 301 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat 361 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta 421 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct 481 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct 541 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct 601 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga 661 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca 721 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata 781 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga 841 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc 901 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg 961 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc 1021 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat 1081 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact 1141 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat 1201 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga 1261 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc 1321 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga 1381 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag 1441 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc 1501 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt 1561 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct 1621 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg 1681 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg 1741 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca 1801 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga 1861 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 1921 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 1981 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt 2041 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 2101 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 2161 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 2221 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 2281 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 2341 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 2401 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 2461 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 2521 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag 2581 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg 2641 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct 2701 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt 2761 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg 2821 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc 2881 ttaattccct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 2941 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 3001 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 3061 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt 3121 tgcaaagatg gataaagttt taaacagaga ggaatctttg cagctaatgg accttctagg 3181 tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca 3241 cagtccccga gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc 3301 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg 3361 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg 3421 ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg 3481 gcccttgcgt gccttgaatt acttccacct ggctgcagta cgtgattctt gatcccgagc 3541 ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc 3601 gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc 3661 ttcgcgcctg tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg 3721 ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc gggccaagat ctgcacactg 3781 gtatttcggt ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt 3841 cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg 3901 gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa 3961 ggctggcccg gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctg 4021 cagggagctc aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac 4081 aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg 4141 cgccgtccag gcacctcgat tagttctcga gcttttggag tacgtcgtct ttaggttggg 4201 gggaggggtt ttatgcgatg gagtttcccc acactgagtg ggtggagact gaagttaggc 4261 cagcttggca cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt 4321 tcattctcaa gcctcagaca gtggttcaaa gtttttttct tccatttcag gtgtcgtgag 4381 gaattctcta gataagccac catggattat aaggatgatg atgataaagg tgattataag 4441 gatgatgatg ataaaggtac caccgagcag cgacgcagcc tgcaagcctt ccaggattat 4501 atccggaaga ccctggaccc tacctacatc ctgagctaca tggccccctg gtttagggag 4561 gaagaggtgc agtatattca ggctgagaaa aacaacaagg gcccaatgga ggctgccaca 4621 ctttttctca agttcctgtt ggagctccag gaggaaggct ggttccgtgg ctttttggat 4681 gccctagacc atgcaggtta ttctggactt tatgaagcca ttgaaagttg ggatttcaaa 4741 aaaattgaaa agttggagga gtatagatta cttttaaaac gtttacaacc agaatttaaa 4801 accagaatta tcccaaccga tatcatttct gatctgtctg aatgtttaat taatcaggaa 4861 tgtgaagaaa ttctacagat ttgctctact aaggggatga tggcaggtgc agagaaattg 4921 gtggaatgcc ttctcagatc agacaaggaa aactggccca aaactttgaa acttgctttg 4981 gagaaagaaa ggaacaagtt cagtgaactg tggattgtag agaaaggtat aaaagatgtt 5041 gaaacagaag atcttgagga taagatggaa acttctgaca tacagatttt ctaccaagaa 5101 gatccagaat gccagaatct tagtgagaat tcatgtccac cttcagaagt gtctgataca 5161 aacttgtaca gcccatttaa accaagaaat taccaattag agcttgcttt gcctgctatg 5221 aaaggaaaaa acacaataat atgtgctcct acaggttgtg gaaaaacctt tgtttcactg 5281 cttatatgtg aacatcatct taaaaaattc ccacaaggac aaaaggggaa agttgtcttt 5341 tttgcgaatc agatcccagt gtatgaacag cagaaatctg tattctcaaa atactttgaa 5401 agacatgggt atagagttac aggcatttct ggagcaacag ctgagaatgt cccagtggaa 5461 cagattgttg agaacaatga catcatcatt ttaactccac agattcttgt gaacaacctt 5521 aaaaagggaa cgattccatc actatccatc tttactttga tgatatttga tgaatgccac 5581 aacactagta aacaacaccc gtacaatatg atcatgttta attatctaga tcagaaactt 5641 ggaggatctt caggcccact gccccaggtc attgggctgg ctgccgcggt tggtgttggg 5701 gatgccaaaa acacagatga agccttggat tatatctgca agctgtgtgc ttctcttgat 5761 gcgtcagtga tagcaacagt caaacacaat ctggaggaac tggagcaagt tgtttataag 5821 ccccagaagt ttttcaggaa agtggaatca cggattagcg acaaatttaa atacatcata 5881 gctcagctga tgagggacac agagagtctg gcaaagagaa tctgcaaaga cctcgaaaac 5941 ttatctcaaa ttcaaaatag ggaatttgga acacagaaat atgaacaatg gattgttaca 6001 gttcagaaag catgcatggt gttccagatg ccagacaaag atgaagagag caggatttgt 6061 aaagccctgt ttttatacac ttcacatttg cggaaatata atgatgccct cattatcagt 6121 gagcatgcac gaatgaaaga tgctctggat tacttgaaag acttcttcag caatgtccga 6181 gcagcaggat tcgatgagat tgagcaagat cttactcaga gatttgaaga aaagctgcag 6241 gaactagaaa gtgtttccag ggatcccagc aatgagaatc ctaaacttga agacctctgc 6301 ttcatcttac aagaagagta ccacttaaac ccagagacaa taacaattct ctttgtgaaa 6361 accagagcac ttgtggacgc tttaaaaaat tggattgaag gaaatcctaa actcagtttt 6421 ctaaaacctg gcatattgac tggacgtggc aaaacaaatc agaacacagg aatgaccctc 6481 ccggcacaga agtgtatatt ggatgcattc aaagccagtg gagatcacaa tattctgatt 6541 gccacctcag ttgctgatga aggcattgac attgcacagt gcaatcttgt catcctttat 6601 gagtatgtgg gcaatgtcat caaaatgatc caaaccagag gcagaggaag agcaagaggt 6661 agcaagtgct tccttctgac tagtaatgct ggtgtaattg aaaaagaaca aataaacatg 6721 tacaaagaaa aaatgatgaa tgactctatt ttacgccttc agacatggga cgaagcagta 6781 tttagggaaa agattctgca tatacagact catgaaaaat tcatcagaga tagtcaagaa 6841 aaaccaaaac ctgtacctga taaggaaaat aaaaaactgc tctgcagaaa gtgcaaagcc 6901 ttggcatgtt acacagctga cgtaagagtg atagaggaat gccattacac tgtgcttgga 6961 gatgctttta aggaatgctt tgtgagtaga ccacatccca agccaaagca gttttcaagt 7021 tttgaaaaaa gagcaaagat attctgtgcc cgacagaact gcagccatga ctggggaatc 7081 catgtgaagt acaagacatt tgagattcca gttataaaaa ttgaaagttt tgtggtggag 7141 gatattgcaa ctggagttca gacgctgtac tcgaagtgga aggactttca ttttgagaag 7201 ataccatttg atccagcaga aatgtccaaa tgatatcagg tcctcaatct tcagctacat 7261 cgatgtcgac ggtacctaga gtgagggtcc ccacctggga cccttgagag tatcaggtct 7321 cccacgtggg agacaagaaa tccctgttta atatttaaac agcagtgttc cccatctggg 7381 tccttgcacc cctcactctg gcctcagccg actgcacagc ggcccctgca tccccttggc 7441 tgtgaggccc ctggacaagc agaggtggcc agagctggga ggcatggccc tggggtccca 7501 cgaatttgct ggggaatctc gtttttcttc ttaagacttt tgggacatgg tttgactccc 7561 gaacatcacc gacgtgtctc ctgtttttct gggtggcctc gggacacctg ccctgccccc 7621 acgagggtca ggactgtgac tctttttagg gccaggcagg tgcctggaca tttgccttgc 7681 tggacgggga ctggggatgt gggagggagc agacaggagg aatcatgtca ggcctgtgtg 7741 tgaaaggaag ctccactgtc accctccacc tcttcacccc ccactcacca gtgtcccctc 7801 cactgtcaca ttgtaactga acttcaggat aataaagtgt ttgcctccaa aaaaaaaaaa 7861 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 7921 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 7981 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 8041 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 8101 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa accgaattca ctggccgtcg ttttacaacg 8161 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt 8221 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag 8281 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc 8341 acaccgcata cgtcaaagca accatagt //