LOCUS       sequence-265282-        8368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses RIG-I with a mutation to motif III in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1074/jbc.M807365200"; journalName: "J Biol Chem"; date: 
            "2009-04-10- 10"; volume: "284"; issue: "15"; pages: "9700-12"
COMMENT     Sequence Label: pEF-Bos-FLAG RIG-I MIII
FEATURES             Location/Qualifiers
     source          1..8368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             join(843..911,912..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        join(2786..2791,2792..2809,2810..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"
     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
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     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
//
